Patent application title: TRAIT IMPROVEMENT IN PLANTS EXPRESSING MYB-RELATED PROTEINS
Inventors:
Colleen M. Marion (San Mateo, CA, US)
Graham J. Hymus (Castro Valley, CA, US)
T. Lynne Reuber (San Mateo, CA, US)
Oliver J. Ratcliffe (Oakland, CA, US)
Assignees:
Mendel Biotechnology, Inc.
IPC8 Class: AC12N1582FI
USPC Class:
800260
Class name: Multicellular living organisms and unmodified parts thereof and related processes method of using a plant or plant part in a breeding process which includes a step of sexual hybridization
Publication date: 2014-02-06
Patent application number: 20140041073
Abstract:
Polynucleotides and polypeptides incorporated into expression vectors are
introduced into plants and were ectopically expressed. These polypeptides
may confer at least one regulatory activity and increased photosynthetic
resource use efficiency, increased yield, greater vigor, greater biomass
as compared to a control plant.Claims:
1. A transgenic plant having greater photosynthetic resource use
efficiency than a control plant; wherein the transgenic plant comprises a
recombinant polynucleotide comprising a promoter that regulates
expression of a polypeptide comprising SEQ ID NO: 2 in a photosynthetic
tissue to a level that is effective in conferring greater photosynthetic
resource use efficiency in the transgenic plant relative to the control
plant; wherein the control plant does not comprise the recombinant
polynucleotide; wherein the promoter does not regulate protein expression
in a constitutive manner; and wherein expression of the polypeptide under
the regulatory control of the promoter confers greater photosynthetic
resource use efficiency in the transgenic plant relative to the control
plant.
2. The transgenic plant of claim 1, wherein the promoter is a photosynthetic tissue-enhanced promoter.
3. The transgenic plant of claim 2, wherein the photosynthetic tissue-enhanced promoter is an RBCS3 promoter, an RBCS4 promoter, an At4g01060 promoter, an Os02g09720 promoter, an Os05g34510 promoter, an Os11g08230 promoter, an Os01g64390 promoter, an Os06g15760 promoter, an Os12g37560 promoter, an Os03g17420 promoter, an Os04g51000 promoter, an Os01g01960 promoter, an Os05g04990 promoter, an Os02g44970 promoter, an Os01g25530 promoter, an Os03g30650 promoter, an Os01g64910 promoter, an Os07g26810 promoter, an Os07g26820 promoter, an Os09g11220 promoter, an Os04g21800 promoter, an Os10g23840 promoter, an Os08g13850 promoter, an Os12g42980 promoter, an Os03g29280 promoter, an Os03g20650 promoter, or an Os06g43920 promoter (SEQ ID NO: 136-159, respectively).
4. The transgenic plant of claim 1, wherein: the recombinant polynucleotide encodes the polypeptide comprising SEQ ID NO: 2; or the polypeptide is encoded by a second polynucleotide and expression of the polypeptide is regulated by a trans-regulatory element.
5. The transgenic plant of claim 1, wherein the transgenic plant has an altered trait that confers the greater photosynthetic resource use efficiency, wherein the altered trait is: (a) increased photosynthetic capacity; and/or (b) increased photosynthetic rate; and/or (c) a decrease in leaf chlorophyll content; and/or (d) a decrease in percentage of nitrogen in leaf dry weight; and/or (e) increased transpiration efficiency; and/or (f) an increase in resistance to water vapor diffusion exerted by leaf stomata; and/or (g) an increase in a rate of reactions responsible for dissipating light energy absorbed by light harvesting antennae as heat; and/or (h) a decrease in the ratio of the carbon isotope 12C to 13C in above-ground biomass; and/or (i) an increase in the total dry weight of above-ground plant material; and/or (j) greater yield than the control plant.
6. The transgenic plant of claim 1, wherein a plurality of the transgenic plants have greater cumulative canopy photosynthesis than the canopy photosynthesis of the same number of the control plants grown under the same conditions and at the same density.
7. The transgenic plant of claim 1, wherein the transgenic plant is selected from the group consisting of a corn, wheat, rice, Setaria, Miscanthus, switchgrass, ryegrass, sugarcane, miscane, barley, sorghum, soy, cotton, canola, rapeseed, Crambe, Camelina, sugar beet, alfalfa, tomato, Eucalyptus, poplar, willow, pine, birch and a woody plant.
8. A method for increasing photosynthetic resource use efficiency in a plant, the method comprising: (a) providing one or more transgenic plants that comprise a recombinant polynucleotide that comprises a photosynthetic tissue-enhanced promoter that regulates a polypeptide comprising SEQ ID NO: 2; and (b) growing the one or more transgenic plants; wherein the photosynthetic tissue-enhanced promoter does not regulate protein expression in a constitutive manner; and wherein expression of the polypeptide in the one or more transgenic plants confers increased photosynthetic resource use efficiency relative to a control plant that does not comprise the recombinant polynucleotide.
9. The method of claim 8, wherein the photosynthetic tissue-enhanced promoter is an RBCS3 promoter, an RBCS4 promoter, an At4g01060 promoter, an Os02g09720 promoter, an Os05g34510 promoter, an Os11g08230 promoter, an Os01g64390 promoter, an Os06g15760 promoter, an Os12g37560 promoter, an Os03g17420 promoter, an Os04g51000 promoter, an Os01g01960 promoter, an Os05g04990 promoter, an Os02g44970 promoter, an Os01g25530 promoter, an Os03g30650 promoter, an Os01g64910 promoter, an Os07g26810 promoter, an Os07g26820 promoter, an Os09g11220 promoter, an Os04g21800 promoter, an Os10g23840 promoter, an Os08g13850 promoter, an Os12g42980 promoter, an Os03g29280 promoter, an Os03g20650 promoter, or an Os06g43920 promoter (SEQ ID NO: 136-159, respectively).
10. The method of claim 8, wherein an expression cassette comprising the recombinant polynucleotide is introduced into a target plant to produce the transgenic plant.
11. The method of claim 8, wherein the transgenic plant has an altered trait that confers the greater photosynthetic resource use efficiency, wherein the altered trait is: (a) increased photosynthetic capacity; and/or (b) increased photosynthetic rate; and/or (c) a decrease in leaf chlorophyll content; and/or (d) a decrease in percentage of nitrogen in leaf dry weight; and/or (e) increased transpiration efficiency; and/or (f) an increase in resistance to water vapor diffusion exerted by leaf stomata; and/or (g) an increase in a rate of reactions responsible for dissipating light energy absorbed by light harvesting antennae as heat; and/or (h) a decrease in the ratio of the carbon isotope 12C to 13C in above-ground biomass; and/or (i) an increase in the total dry weight of above-ground plant material; and/or (j) greater yield than the control plant.
12. The method of claim 8, wherein the transgenic plant is selected for having the increased photosynthetic resource use efficiency relative to the control plant.
13. The method of claim 12, wherein the plant is selected for having the greater yield relative to the control plant.
14. The method of claim 8, wherein a plurality of the transgenic plants have greater cumulative canopy photosynthesis than the canopy photosynthesis of the same number of the control plants grown under the same conditions and at the same density.
15. The method of claim 8, the method steps further including: crossing the target plant with itself, a second plant from the same line as the target plant, a non-transgenic plant, a wild-type plant, or a transgenic plant from a different line of plants, to produce a transgenic seed.
16. A method for producing and selecting a crop plant with greater yield than a control plant, the method comprising: (a) providing one or more transgenic plants that comprise a recombinant polynucleotide that comprises photosynthetic tissue-enhanced promoter that regulates a polypeptide comprising SEQ ID NO: 2, wherein the photosynthetic tissue-enhanced promoter does not regulate protein expression in a constitutive manner; (b) growing a plurality of the transgenic plants; and (c) selecting a transgenic plant that: has greater photosynthetic resource use efficiency than the control plant, wherein the control plant does not comprise the recombinant polynucleotide; and/or comprises the recombinant polynucleotide; wherein expression of the polypeptide in the selected transgenic plant confers the greater yield of the selected transgenic plant relative to the control plant.
17. The method of claim 16, the method steps further including: (d) crossing the selected transgenic plant with itself, a second plant from the same line as the selected transgenic plant, a non-transgenic plant, a wild-type plant, or a transgenic plant from a different line of plants, to produce a transgenic seed.
18. The method of claim 16, wherein the transgenic plant is selected for having the increased photosynthetic resource use efficiency relative to the control plant.
19. The method of claim 16, wherein a plurality of the selected transgenic plants have greater cumulative canopy photosynthesis than the canopy photosynthesis of the same number of the control plants grown under the same conditions and at the same density.
20. The method of claim 16, wherein the selected transgenic plant has an altered trait that confers the greater photosynthetic resource use efficiency, wherein the altered trait is: (a) increased photosynthetic capacity; and/or (b) increased photosynthetic rate; and/or (c) a decrease in leaf chlorophyll content; and/or (d) a decrease in percentage of nitrogen in leaf dry weight; and/or (e) increased transpiration efficiency; and/or (f) an increase in resistance to water vapor diffusion exerted by leaf stomata; and/or (g) an increase in a rate of reactions responsible for dissipating light energy absorbed by light harvesting antennae as heat; and/or (h) a decrease in the ratio of the carbon isotope 12C to 13C in above-ground biomass; and/or (i) an increase in the total dry weight of above-ground plant material.
Description:
[0001] This application claims the benefit of copending U.S. Provisional
Application No. 61/679,320, filed Aug. 3, 2012, the entire contents of
which are hereby incorporated by reference.
FIELD OF THE INVENTION
[0002] The present invention relates to plant genomics and plant improvement.
BACKGROUND OF THE INVENTION
[0003] A plant's phenotypic characteristics that enhance photosynthetic resource use efficiency may be controlled through a number of cellular processes. One important way to manipulate that control is by manipulating the characteristics or expression of regulatory proteins, proteins that influence the expression of a particular gene or sets of genes. For example, transformed or transgenic plants that comprise cells with altered levels of at least one selected regulatory polypeptide may possess advantageous or desirable traits, and strategies for manipulating traits by altering a plant cell's regulatory polypeptide content or expression level can result in plants and crops with commercially valuable properties. Examples of such trait manipulation are increased canopy photosynthesis, nitrogen use efficiency, or water use efficiency, as considered below.
[0004] Increasing Canopy Photosynthesis to Increase Crop Yield.
[0005] Recent studies by crop physiologists have provided evidence that crop-canopy photosynthesis is correlated with crop yield, and that increasing canopy photosynthesis can increase crop yield (Long et al., 2006. Plant Cell Environ. 29:315-33; Murchie et al., 2009 New Phytol. 181:532-552; Zhu et al., 2010. Ann. Rev. Plant Biol. 61:235-261). Two overlapping strategies for increasing canopy photosynthesis have been proposed. The first recognizes that there exists great potential to increase canopy photosynthesis by improving multiple discrete reactions that currently limit photosynthetic capacity (reviewed in Zhu et al., 2010. supra). The second focuses upon improving plant physiological status during environmental conditions that limit the realization of photosynthetic capacity. It is important to distinguish this second goal from recent industry and academic screening for genes to improve stress tolerance. Arguably, these efforts may have identified genes that improve plant physiological status during severe stresses not typically experienced on productive acres (Jones, 2007. J. Exp. Bot. 58:119-130; Passioura, 2007. J. Exp. Bot. 58:113-117). In contrast, improving the resource use efficiency with which photosynthesis operates relative to the availability of key resources of water, nitrogen and light, is thought to be more appropriate for improving yield on productive acres (Long et al., 1994. Ann. Rev. Plant Physiol. Plant Molec. Biol. 45:633-662; Morison et al., 2008. Philosophical Transactions of the Royal Society B: Biological Sciences 363:639-658; Passioura, 2007, supra).
[0006] Increasing Nitrogen Use Efficiency (NUE) to Increase Crop Yield.
[0007] There has been a large increase in food productivity over the past 50 years causing a decrease in world hunger despite a significant increase in population (Godfray et al., 2010. Science 327:812-818). A significant contribution to this increased yield was a 20-fold increase in the application of nitrogen fertilizers (Glass, 2003. Crit. Rev. Plant Sci. 22:453-470). About 85 million to 90 million metric tons of nitrogen are applied annually to soil, and this application rate is expected to increase to 240 million metric tons by 2050 (Good et al., 2004. Trends Plant Sci. 9:597-605). However, plants use only 30 to 40% of the applied nitrogen and the rest is lost through a combination of leaching, surface run-off, denitrification, volatilization, and microbial consumption (Frink et al., 1999. Proc. Natl. Acad. Sci. USA 96:1175-1180; Glass, 2003, supra; Good et al., 2004, supra; Raun and Johnson, 1999. Agron. J. 91:357-363). The loss of more than 60% of applied nitrogen can have serious environmental effects, such as groundwater contamination, anoxic coastal zones, and conversion to greenhouse gases. In addition, while most fertilizer components are mined (such as phosphates), inorganic nitrogen is derived from the energy intensive conversion of gaseous nitrogen to ammonia. Thus, the addition of nitrogen fertilizer is typically the highest single input cost for many crops, and since its production is energy intensive, the cost is dependent on the price of energy (Rothstein, 2007. Plant Cell 19:2695-2699). With an increasing demand for food from an increasing human population, agriculture yields must be increased at the same time as dependence on applied fertilizers is decreased. Therefore, to minimize nitrogen loss, reduce environmental pollution, and decrease input cost, it is crucial to develop crop varieties with higher nitrogen use efficiency (Garnett et al., 2009. Plant Cell Environ. 32:1272-1283; Hirel et al., 2007. J. Exp. Bot. 58:2369-2387; Lea and Azevedo, 2007. Ann. Appl. Biol. 151:269-275; Masclaux-Daubresse et al., 2010. Ann. Bot. 105:1141-1157; Moll et al., 1982. Agron. J. 74:562-564; Sylvester-Bradley and Kindred, 2009. J. Exp. Bot. 60:1939-1951).
[0008] Improving Water Use Efficiency (WUE) to Improve Yield.
[0009] Freshwater is a limited and dwindling global resource; therefore, improving the efficiency with which food and biofuel crops use water is a prerequisite for maintaining and improving yield (Karaba et al., 2007. Proc. Natl. Acad. Sci. USA. 104:15270-15275). WUE can be used to describe the relationship between water use and crop productivity over a range of time integrals. The basic physiological definition of WUE equates the ratio of photosynthesis (A) to transpiration (T) at a given moment in time, also referred to as transpiration efficiency. However, the WUE concept can be scaled significantly, for example, over the complete lifecycle of a crop, where biomass or yield can be expressed per cumulative total of water transpired from the canopy. Thus far, the engineering of major field crops for improved WUE with single genes has not yet been achieved (Karaba et al., 2007. supra). Regardless, increased yields of wheat cultivars bred for increased transpiration efficiency (the ratio of photosynthesis to transpiration) have provided important support for the proposition that crop yield can be increased over broad acres through improvement in crop water-use efficiency (Condon et al., 2004. J. Exp. Bot. 55:2447-2460).
[0010] With these needs in mind, new technologies for yield enhancement are required. In this disclosure, a phenotypic screening platform that directly measures photosynthetic capacity, water-use efficiency, and nitrogen use efficiency of mature plants was used to discover advantageous properties conferred by ectopic expression of the described regulatory proteins in plants.
SUMMARY
[0011] The instant description is directed to a transgenic plant or plants that have increased photosynthetic resource use efficiency with respect to a control plant. In this regard, the transgenic plant or plants comprise a first recombinant polynucleotide comprising a promoter of interest. The choice of promoter may include a constitutive promoter or a promoter with enhanced activity in a tissue capable of photosynthesis (also referred to herein as a "photosynthetic promoter" or a "photosynthetic tissue-enhanced promoter") such as a leaf tissue or other green tissue. Examples of photosynthetic promoters include for example, an RBCS3 promoter (SEQ ID NO: 133), an RBCS4 promoter (SEQ ID NO: 134) or others such as the At4g01060 (also referred to as "G682") promoter (SEQ ID NO: 135), the latter regulating expression in a guard cell. The promoter regulates a polypeptide that is encoded by the first recombinant polynucleotide or by a second (or target) recombinant polynucleotide (in which case expression of the polypeptide may be regulated by a trans-regulatory element). The promoter may also regulate expression of a polypeptide to an effective level of expression in a photosynthetic tissue, that is, to a level that, as a result of expression of the polypeptide to that level, improves photosynthetic resource use efficiency in a transgenic plant relative to a control plant. The first polynucleotide may comprise the promoter and also encode the polypeptide or alternatively, the first polypeptide may comprise the promoter and drive expression of the polypeptide which is encoded by the second recombinant polynucleotide. In a preferred embodiment, the polypeptide comprises SEQ ID NO: 2 or a sequence that is paralogous or orthologous to SEQ ID NO: 2, being structurally-related to SEQ ID NO: 2 and having a function similar to SEQ ID NO: 2 as described herein. Expression of the polypeptide under the regulatory control of the constitutive or the leaf-enhanced or photosynthetic tissue-enhanced promoter in the transgenic plant confers greater photosynthetic resource use efficiency to the transgenic plants, and may ultimately increase yield that may be obtained from the plants.
[0012] The instant description also pertains to methods for increasing photosynthetic resource use efficiency in, or increasing yield from, a plant or plants including: the method conducted by growing a transgenic plant comprising and/or transformed with an expression cassette comprising the first recombinant polynucleotide that comprises a constitutive promoter or a promoter expressed in photosynthetic tissue, which may be a leaf-enhanced or green tissue-enhanced promoter, such as for example, the RBCS3, RBCS4 or At4g01060 promoters, or another photosynthetic tissue-enhanced promoter, for example, such a promoter found in the sequence listing or in Table 4. Said promoter regulates expression of a polypeptide that comprises SEQ ID NO: 2, or a polypeptide sequence within the MYB19 clade (recombinant polynucleotides encoding MYB19 clade polypeptides are described in the following paragraphs (a)-(c), and exemplary polypeptides within the clade are described in the following paragraphs (d)-(f) and are shown in FIG. 1 and FIGS. 2A-2I).
[0013] The first or second recombinant polynucleotide encoding a MYB19 clade polypeptide may include:
[0014] (a) nucleic acid sequences that are at least 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% or 96%, 97%, 98%, 99%, or about 100% identical to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33; and/or
[0015] (b) nucleic acid sequences that encode polypeptide sequences that are at least 40%, increasing by increments of 1% to about 100% identical in their amino acid sequences to the entire length of any of SEQ ID NOs: 2n, where n=1-17 (that is, SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34); and/or
[0016] (c) nucleic acid sequences that hybridize under stringent conditions (e.g., hybridization followed by one, two, or more wash steps of 6×SSC and 65° C. for ten to thirty minutes per step) to any of SEQ ID NOs: 2n-1, where n=1-17 (that is, SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33).
[0017] The MYB19 clade polypeptides may include:
[0018] (d) polypeptide sequences encoded by the nucleic acid sequences of (a), (b) and/or (c); and/or
[0019] (e) polypeptide sequences that have at least 40% identity increasing by increments of 1% to about 100% identity to SEQ ID NO: 2 or to SEQ ID NOs: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, or 34, and/or at least 69% identity increasing by increments of 1% to about 100% identity to the first Myb DNA binding domain of SEQ ID NO: 2 (`Myb DNA binding domain 1`) or SEQ ID NOs: 61-77, and/or at least 72% identity increasing by increments of 1% to about 100% identity to the second Myb DNA binding domain (`Myb DNA binding domain 2`) of SEQ ID NO: 2 or SEQ ID NOs: 95-111; and/or
[0020] (f) polypeptide sequences that comprise a subsequence that are at least 95%, 96%, 97%, 98%, 99%, or about 100% identical to a consensus sequence of SEQ ID NO: 129 or 130.
[0021] "Increasing by increments of 1% to about 100% identity" in paragraphs (b) and (e) refers to at least: 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 90%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% or 96%, 97%, 98%, 99%, or about 100% amino acid identity to SEQ ID NO: 2 or to SEQ ID NOs: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, or 34; or
[0022] at least: 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 90%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% or 96%, 97%, 98%, or at least 99%, or about 100% identity to any the first Myb DNA binding domains of SEQ ID NOs: 61 to 77; or
[0023] at least: 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% or 96%, 97%, 98%, 99%, or about 100% identity to any of the second Myb DNA binding domains of SEQ ID NOs: 95 to 111.
[0024] Expression of these MYB19 clade polypeptides in the transgenic plant may confer increased photosynthetic resource use efficiency relative to a control plant. The transgenic plant may be selected for increased photosynthetic resource use efficiency or greater yield relative to the control plant. The transgenic plant may also be crossed with itself, a second plant from the same line as the transgenic plant, a non-transgenic plant, a wild-type plant, or a transgenic plant from a different line of plants, to produce a transgenic seed.
[0025] The instant description also pertains to methods for producing and selecting a crop plant with a greater yield than a control plant, the method comprising producing a transgenic plant by introducing into a target plant a recombinant polynucleotide that comprises a promoter, such as a leaf- or photosynthetic tissue-enhanced promoter that regulates a polypeptide encoded by the recombinant polynucleotide or a second recombinant polynucleotide, wherein the polypeptide comprises SEQ ID NO: 2. A plurality of the transgenic plants are then grown, and a transgenic plant is selected that produces greater yield or has greater photosynthetic resource use efficiency than a control plant. The expression of the polypeptide in the selected transgenic plant confers the greater photosynthetic resource use efficiency and/or greater yield relative to the control plant. Optionally, the selected transgenic plant may be crossed with itself, a second plant from the same line as the transgenic plant, a non-transgenic plant, a wild-type plant, or a transgenic plant from a different line of plants, to produce a transgenic seed. A plurality of the selected transgenic plants will generally have greater cumulative canopy photosynthesis than the canopy photosynthesis of an identical number of the control plants.
[0026] The transgenic plant(s) described herein and produced by the instantly described methods may also possess one or more altered traits that result in greater photosynthetic resource use efficiency. The altered trait may include: increased photosynthetic capacity; a decrease in leaf chlorophyll content; a decrease in percentage of nitrogen in leaf dry weight; increased transpiration efficiency; an increase in resistance to water vapor diffusion exerted by leaf stomata; an increase in a rate of reactions responsible for dissipating light energy absorbed by light harvesting antennae as heat; a decrease in the ratio of the carbon isotope 12C to 13C in above-ground biomass; and/or an increase in the total dry weight of above-ground plant material.
[0027] At least one advantage of greater photosynthetic resource use efficiency is that the transgenic plant, or a plurality of the transgenic plants, will have greater cumulative canopy photosynthesis than the canopy photosynthesis of an identical number of the control plants, or produce greater yield than an identical number of the control plants. A wide variety of transgenic plants are envisioned, including corn, wheat, rice, Setaria, Miscanthus, Setaria switchgrass, ryegrass, sugarcane, miscane, barley, sorghum, soy, cotton, canola, rapeseed, Crambe, Camelina, sugar beet, alfalfa, tomato, Eucalyptus, poplar, willow, pine, birch and other woody plants.
[0028] The instant description also pertains to expression vectors that comprise a recombinant polynucleotide that comprises a promoter expressed in photosynthetic tissue, for example a leaf- or green tissue-enhanced promoter including the RBCS3, RBCS4, or At4g01060 promoters (SEQ ID NOs: 133-135), or another photosynthetic tissue-enhanced promoter, for example, such a promoter found in the sequence listing or in Table 4 (e.g., SEQ ID NOs: 136-159), and a subsequence that encodes a polypeptide comprising SEQ ID NO: 2, or, alternatively, two expression constructs, one of which encodes a promoter such as a leaf-enhanced promoter or other photosynthetic tissue-enhanced promoter, and the second encodes the polypeptide comprising SEQ ID NO: 2. In either instance, whether the polypeptide is encoded by the first or second expression constructs, the promoter regulates expression of the polypeptide comprising SEQ ID NO: 2 by being responsible for production of cis- or trans-regulatory elements, respectively.
[0029] In the above paragraphs, the control plant may be exemplified by a plant of the same species as the plant comprising the recombinant polynucleotide, but the control plant does not comprise the recombinant polynucleotide (containing the promoter and possibly encoding the polypeptide) or the second recombinant polynucleotide.
BRIEF DESCRIPTION OF THE SEQUENCE LISTING AND DRAWINGS
[0030] The Sequence Listing provides exemplary polynucleotide and polypeptide sequences of the instant description. The traits associated with the use of the sequences are included in the Examples.
[0031] Incorporation of the Sequence Listing.
[0032] The Sequence Listing provides exemplary polynucleotide and polypeptide sequences. The copy of the Sequence Listing, being submitted electronically with this patent application, provided under 37 CFR §1.821-1.825, is a read-only memory computer-readable file in ASCII text format. The Sequence Listing is named "MBI-0203P_ST25.txt", the electronic file of the Sequence Listing was created on Aug. 3, 2012, and is 222,861 bytes in size (217 kilobytes in size as measured in MS-WINDOWS). The Sequence Listing is herein incorporated by reference in its entirety.
[0033] In FIG. 1, a phylogenetic tree of the MYB19 (also referred to as G1309) clade members and related full length proteins were constructed using TreeBeST (Ruan et al., 2008. Nucleic Acids Res. 36 (suppl. 1): D735-D740) using the best command to identify the best tree from maximum likelihood and neighbor joining methods. The MYB19 clade members appear in the large box with the solid line boundary. MYB19 appears in the oval. An ancestral sequence of MYB19 and closely-related sequences is represented by the node of the tree indicated by the arrow "A" in FIG. 1. MYB19 clade members are considered those proteins that descended from ancestral sequence "A", including the exemplary sequences shown in this figure that are bounded by LOC_Os04g45020.1 and Solyc03g025870.2.1 (indicated by the box around these sequences). A related clade is represented by the node indicated by arrow "B".
[0034] FIGS. 2A-2I show an alignment of the MYB19 clade and related proteins which appear in the boxes with the solid line boundaries. The alignment was generated with MUSCLE v3.8.31 (Edgar (2004) Nucleic Acids Res. 32:1792-1797) with default parameters. SEQ ID NOs: appear in parentheses after each Gene Identifier (GID). The conserved first and second Myb DNA binding domains appear in boxes with the dashed line boundaries. The conserved residues within the clade are shown in the last rows of FIGS. 2B-2F and are presented as SEQ ID NOs: 129 (underlined), 130 (double underlined) and 160. SEQ ID NOs: 129 and 130 share the triple underlined Glu residue in FIG. 2C.
[0035] FIG. 3 presents a plot of photosynthetic capacity at growth temperature, showing increased light saturated photosynthesis (Asat) over a range of leaf, sub-stomatal CO2 concentration (Ci), in five MYB19 overexpression lines, compared to a control line. Data were collected over a range of Ci over which the activity of Rubisco is known to limit Asat. The solid line shown is a regression fitted to the data for the control line only. All data are the means±1 standard error for data collected on at least nine replicate plants for each line.
[0036] FIG. 4 presents a plot of photosynthetic capacity at growth temperature showing increased Asat over a range of leaf, sub-stomatal Ci in five MYB19 overexpression lines, compared to a control line. Data were collected over a range of Ci over which the capacity to regenerate RuBP is known to limit Asat. The solid line shown is a regression fitted to the data for the control line only. All data are the means±1 standard error for data collected on at least nine replicate plants for each line.
LEGEND FOR FIG. 3 AND FIG. 4
[0037] control
[0038] ∘ Line 2
[0039] ⋄ Line 3
[0040] Δ Line 6
[0041] quadrature Line 7
[0042] Line 8
DETAILED DESCRIPTION
[0043] The present description relates to polynucleotides and polypeptides for modifying phenotypes of plants, particularly those associated with increased photosynthetic resource use efficiency and increased yield with respect to a control plant (for example, a wild-type plant). Throughout this disclosure, various information sources are referred to and/or are specifically incorporated. The information sources include scientific journal articles, patent documents, textbooks, and internet entries. While the reference to these information sources clearly indicates that they can be used by one of skill in the art, each and every one of the information sources cited herein are specifically incorporated in their entirety, whether or not a specific mention of "incorporation by reference" is noted. The contents and teachings of each and every one of the information sources can be relied on and used to make and use embodiments of the instant description.
[0044] As used herein and in the appended claims, the singular forms "a", "an", and "the" include the plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to "a host cell" includes a plurality of such host cells, and a reference to "a plant" is a reference to one or more plants, and so forth.
[0045] A "recombinant polynucleotide" is a polynucleotide that is not in its native state, e.g., the polynucleotide comprises a nucleotide sequence not found in nature, or the polynucleotide is in a context other than that in which it is naturally found, e.g., separated from nucleotide sequences with which it typically is in proximity in nature, or adjacent (or contiguous with) nucleotide sequences with which it typically is not in proximity For example, the sequence at issue can be cloned into a vector, or otherwise recombined with one or more additional nucleic acid.
[0046] A "polypeptide" is an amino acid sequence comprising a plurality of consecutive polymerized amino acid residues e.g., at least about 15 consecutive polymerized amino acid residues. In many instances, a polypeptide comprises a polymerized amino acid residue sequence that is a regulatory polypeptide or a domain or portion or fragment thereof. Additionally, the polypeptide may comprise: (i) a localization domain; (ii) an activation domain; (iii) a repression domain; (iv) an oligomerization domain; (v) a protein-protein interaction domain; (vi) a DNA-binding domain; or the like. The polypeptide optionally comprises modified amino acid residues, naturally occurring amino acid residues not encoded by a codon, or non-naturally occurring amino acid residues.
[0047] "Protein" refers to an amino acid sequence, oligopeptide, peptide, polypeptide or portions thereof whether naturally occurring or synthetic.
[0048] A "recombinant polypeptide" is a polypeptide produced by translation of a recombinant polynucleotide. A "synthetic polypeptide" is a polypeptide created by consecutive polymerization of isolated amino acid residues using methods well known in the art. An "isolated polypeptide," whether a naturally occurring or a recombinant polypeptide, is more enriched in (or out of) a cell than the polypeptide in its natural state in a wild-type cell, e.g., more than about 5% enriched, more than about 10% enriched, or more than about 20%, or more than about 50%, or more, enriched, i.e., alternatively denoted: 105%, 110%, 120%, 150% or more, enriched relative to wild type standardized at 100%. Such an enrichment is not the result of a natural response of a wild-type plant. Alternatively, or additionally, the isolated polypeptide is separated from other cellular components with which it is typically associated, e.g., by any of the various protein purification methods herein.
[0049] "Identity" or "similarity" refers to sequence similarity between two polynucleotide sequences or between two polypeptide sequences, with identity being a more strict comparison. The phrases "percent identity" and "% identity" refer to the percentage of sequence similarity found in a comparison of two or more polynucleotide sequences or two or more polypeptide sequences. "Sequence similarity" refers to the percent similarity in base pair sequence (as determined by any suitable method) between two or more polynucleotide sequences. Two or more sequences can be anywhere from 0-100% similar or identical, or any integer value between 0-100%. Identity or similarity can be determined by comparing a position in each sequence that may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same nucleotide base or amino acid, then the molecules are identical at that position. A degree of similarity or identity between polynucleotide sequences is a function of the number of identical, matching or corresponding nucleotides at positions shared by the polynucleotide sequences. A degree of identity of polypeptide sequences is a function of the number of identical amino acids at corresponding positions shared by the polypeptide sequences. A degree of homology or similarity of polypeptide sequences is a function of the number of amino acids at corresponding positions shared by the polypeptide sequences. The fraction or percentage of components in common is related to the homology or identity between the sequences. Alignments such as those of FIGS. 2A-2I may be used to identify conserved domains and relatedness within these domains. An alignment may suitably be determined by means of computer programs known in the art, such as MACVECTOR software, (1999; Accelrys, Inc., San Diego, Calif.).
[0050] "Homologous sequences" refers to polynucleotide or polypeptide sequences that are similar due to common ancestry and sequence conservation. The terms "ortholog" and "paralog" are defined below in the section entitled "Orthologs and Paralogs". In brief, orthologs and paralogs are evolutionarily related genes that have similar sequences and functions. Orthologs are structurally related genes in different species that are derived by a speciation event. Paralogs are structurally related genes within a single species that are derived by a duplication event.
[0051] "Functional homologs" are polynucleotide or polypeptide sequences, including orthologs and paralogs, that are similar due to common ancestry and sequence conservation and have identical or similar function at the catalytic, cellular, or organismal levels. The presently disclosed MYB19 clade polypeptides are "functionally-related and/or closely-related" by having descended from a common ancestral sequence (see the node shown by arrow A in FIG. 1), and/or by being sufficiently similar to the sequences and domains listed in Tables 2 or 3 that they confer the same function to plants of increased photosynthetic resource use efficiency and associated improved plant vigor, quality, yield, size, and/or biomass.
[0052] Functionally-related and/or closely-related polypeptides may be created artificially, semi-synthetically, or may occur naturally by having descended from the same ancestral sequence as the disclosed MYB19-related sequences, where the polypeptides have the function of conferring increased photosynthetic resource use efficiency to plants.
[0053] "Conserved domains" are recurring units in molecular evolution, the extents of which can be determined by sequence and structure analysis. A "conserved domain" or "conserved region" as used herein refers to a region in heterologous polynucleotide or polypeptide sequences where there is a relatively high degree of sequence identity between the distinct sequences. Conserved domains contain conserved sequence patterns or motifs that allow for their detection in, and identification and characterization of, polypeptide sequences. A Myb or Myb-like domain is an example of a conserved domain.
[0054] A transgenic plant is expected to have improved or increased photosynthetic resource use efficiency relative to a control plant when the transgenic plant is transformed with a recombinant polynucleotide encoding any of the listed sequences or another MYB19 clade sequence, or when the transgenic plant contains or expresses a MYB19 clade sequence.
[0055] The terms "highly stringent" or "highly stringent condition" refer to conditions that permit hybridization of DNA strands whose sequences are highly complementary, wherein these same conditions exclude hybridization of significantly mismatched DNAs. Polynucleotide sequences capable of hybridizing under stringent conditions with the polynucleotides of the present description may be, for example, variants of the disclosed polynucleotide sequences, including allelic or splice variants, or sequences that encode orthologs or paralogs of presently disclosed polypeptides. Nucleic acid hybridization methods are disclosed in detail by Kashima et al., 1985. Nature 313: 402-404; Sambrook et al., 1989. Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., and by Haymes et al., 1985. Nucleic Acid Hybridization: A Practical Approach, IRL Press, Washington, D.C., which references are incorporated herein by reference.
[0056] In general, stringency is determined by the temperature, ionic strength, and concentration of denaturing agents (e.g., formamide) used in a hybridization and washing procedure (for a more detailed description of establishing and determining stringency, see the section "Identifying Polynucleotides or Nucleic Acids by Hybridization", below). The degree to which two nucleic acids hybridize under various conditions of stringency is correlated with the extent of their similarity. Thus, similar nucleic acid sequences from a variety of sources, such as within a plant's genome (as in the case of paralogs) or from another plant (as in the case of orthologs) that may perform similar functions can be isolated on the basis of their ability to hybridize with known related polynucleotide sequences. Numerous variations are possible in the conditions and means by which nucleic acid hybridization can be performed to isolate related polynucleotide sequences having similarity to sequences known in the art and are not limited to those explicitly disclosed herein. Such an approach may be used to isolate polynucleotide sequences having various degrees of similarity with disclosed polynucleotide sequences, such as, for example, encoded regulatory polypeptides also having at least 40% identity to SEQ ID NO: 2, and/or 69% identity to the first Myb DNA binding domain of SEQ ID NO: 2, and/or 72% identity to the second Myb DNA binding domain of SEQ ID NO: 2, increasing by steps of 1% to about 100%, identity with the conserved domains of disclosed sequences (see, for example, Table 2 showing MYB19 clade polypeptides having at least 69%, 70%, 72%, 73%, 75%, 76%, 77%, 78%, 80%, 85%, or about 100% amino acid identity with the first Myb DNA binding domain of SEQ ID NO: 2, and/or, in Table 3, at least 72%, 74%, 76%, 79%, 81%, 88% or about 100% amino acid identity with the second Myb DNA binding domain of SEQ ID NO: 2).
[0057] "Fragment", with respect to a polynucleotide, refers to a clone or any part of a polynucleotide molecule that retains a usable, functional characteristic. Useful fragments include oligonucleotides and polynucleotides that may be used in hybridization or amplification technologies or in the regulation of replication, transcription or translation. A "polynucleotide fragment" refers to any subsequence of a polynucleotide, typically, of at least about nine consecutive nucleotides, preferably at least about 30 nucleotides, more preferably at least about 50 nucleotides, of any of the sequences provided herein. Exemplary polynucleotide fragments are the first 60 consecutive nucleotides of the polynucleotides listed in the Sequence Listing. Exemplary fragments also include fragments that comprise a region that encodes an conserved domain of a polypeptide. Exemplary fragments also include fragments that comprise a conserved domain of a polypeptide. Exemplary fragments include fragments that comprise an conserved domain of a polypeptide, for example, amino acid residues 17-77 or 70-112 of MYB19 (SEQ ID NO: 2), or the amino acid residues of the domains listed in Tables 2 or 3, or SEQ ID NO: 61-77 or 95-111.
[0058] Fragments may also include subsequences of polypeptides and protein molecules, or a subsequence of the polypeptide. Fragments may have uses in that they may have antigenic potential. In some cases, the fragment or domain is a subsequence of the polypeptide which performs at least one biological function of the intact polypeptide in substantially the same manner, or to a similar extent, as does the intact polypeptide. For example, a polypeptide fragment can comprise a recognizable structural motif or functional domain such as a DNA-binding site or domain that binds to a DNA promoter region, an activation domain, or a domain for protein-protein interactions, and may initiate transcription. Fragments can vary in size from as few as three amino acid residues to the full length of the intact polypeptide, but are preferably at least about 30 amino acid residues in length and more preferably at least about 60 amino acid residues in length.
[0059] Fragments may also refer to a functional fragment of a promoter region. For example, a recombinant polynucleotide capable of modulating transcription in a plant may comprise a nucleic acid sequence with similarity to, or a percentage identity to, a promoter region exemplified by a promoter sequence provided in the Sequence Listing (also see promoters listed in Example I), a fragment thereof, or a complement thereof, wherein the nucleic acid sequence, or the fragment thereof, or the complement thereof, regulates expression of a polypeptide in a plant cell.
[0060] The term "plant" includes whole plants, shoot vegetative organs/structures (for example, leaves, stems and tubers), roots, flowers and floral organs/structures (for example, bracts, sepals, petals, stamens, carpels, anthers and ovules), seed (including embryo, endosperm, and seed coat) and fruit (the mature ovary), plant tissue (for example, vascular tissue, ground tissue, and the like) and cells (for example, guard cells, egg cells, and the like), and progeny of same. The class of the plants that can be transformed using the methods provided of the instant description is generally as broad as the class of higher and lower plants amenable to transformation techniques, including angiosperms (monocotyledonous and dicotyledonous plants), gymnosperms, ferns, horsetails, psilophytes, lycophytes, and bryophytes.
[0061] A "control plant" as used in the present description refers to a plant cell, seed, plant component, plant tissue, plant organ or whole plant used to compare against transgenic or genetically modified plant for the purpose of identifying an enhanced phenotype in the transgenic or genetically modified plant. A control plant may in some cases be a transgenic plant line that comprises an empty vector or marker gene, but does not contain the recombinant polynucleotide of the present description that is expressed in the transgenic or genetically modified plant being evaluated. In general, a control plant is a plant of the same line or variety as the transgenic or genetically modified plant being tested. A suitable control plant would include a genetically unaltered or non-transgenic plant of the parental line used to generate a transgenic plant herein.
[0062] A "transgenic plant" refers to a plant that contains genetic material not found in a wild-type plant of the same species, variety or cultivar. The genetic material may include a transgene, an insertional mutagenesis event (such as by transposon or T-DNA insertional mutagenesis), an activation tagging sequence, a mutated sequence, a homologous recombination event or a sequence modified by chimeraplasty. Typically, the foreign genetic material has been introduced into the plant by human manipulation, but any method can be used as one of skill in the art recognizes.
[0063] A transgenic line or transgenic plant line refers to the progeny plant or plants deriving from the stable integration of heterologous genetic material into a specific location or locations within the genome of the original transformed cell.
[0064] A transgenic plant may contain an expression vector or cassette. The expression cassette typically comprises a polypeptide-encoding sequence operably linked (i.e., under regulatory control of) to appropriate inducible, tissue-enhanced, tissue-specific, or constitutive regulatory sequences that allow for the controlled expression of the polypeptide. The expression cassette can be introduced into a plant by transformation or by breeding after transformation of a parent plant. A plant refers to a whole plant as well as to a plant part, such as seed, fruit, leaf, or root, plant tissue, plant cells or any other plant material, e.g., a plant explant, as well as to progeny thereof, and to in vitro systems that mimic biochemical or cellular components or processes in a cell.
[0065] "Germplasm" refers to a genetic material or a collection of genetic resources for an organism from an individual plan, a group of related individual plants (for example, a plant line, a plant variety or a plant family), or a clone derived from a plant line, plant variety, plant species, or plant culture.
[0066] A constitutive promoter is active under most environmental conditions, and in most plant parts. Regulation of protein expression in a constitutive manner refers to the control of expression of a gene and/or its encoded protein in all tissues regardless of the surrounding environment or development stage of the plant.
[0067] Alternatively, expression of the disclosed or listed polypeptides may be under the regulatory control of a promoter that is not a constitutive promoter. For example, tissue-enhanced (also referred to as tissue-preferred), tissue-specific, cell type-specific, and inducible promoters constitute non-constitutive promoters; that is, these promoters do not regulate protein expression in a constitutive manner. Tissue-enhanced or tissue-preferred promoters facilitate expression of a gene and/or its encoded protein in specific tissue(s) and generally, although perhaps not completely, do not express the gene and/or protein in all other tissues of the plant, or do so to a much lesser extent. Promoters under developmental control include promoters that preferentially initiate transcription in certain tissues, such as xylem, leaves, roots, or seeds. Such promoters are examples of tissue-enhanced or tissue-preferred promoters (see U.S. Pat. No. 7,365,186). Tissue-specific promoters generally confine transgene expression to a single plant part, tissue or cell-type, although many such promoters are not perfectly restricted in their expression and their regulatory control is more properly described as being "tissue-enhanced" or "tissue-preferred". Tissue-enhanced promoters primarily regulate transgene expression in a limited number of plant parts, tissues or cell-types, and the latter type of promoters causes the expression of proteins to be overwhelming restricted to a few particular tissues, plant parts, or cell types. An example of a tissue-enhanced promoter is a "photosynthetic tissue-enhanced promoter", for which the promoter preferentially regulates gene or protein expression in photosynthetic tissues (e.g., leaves, cotyledons, stems, etc.). Tissue-enhanced promoters can be found upstream and operatively linked to DNA sequences normally transcribed in higher levels in certain plant tissues or specifically in certain plant tissues, respectively. "Cell-enhanced", "tissue-enhanced", or "tissue-specific" regulation thus refer to the control of gene or protein expression, for example, by a promoter, which drives expression that is not necessarily totally restricted to a single type of cell or tissue, but where expression is elevated in particular cells or tissues to a greater extent than in other cells or tissues within the organism, and in the case of tissue-specific regulation, in a manner that is primarily elevated in a specific tissue. Tissue-enhanced or preferred promoters have been described in, for example, U.S. Pat. No. 7,365,186, or U.S. Pat. No. 7,619,133.
[0068] Another example of a promoter that is not a constitutive promoter is a "condition-enhanced" promoter, the latter term referring to a promoter that activates a gene in response to a particular environmental stimulus. This may include, for example, an abiotic stress, infection caused by a pathogen, light treatment, etc., and a condition-enhanced promoter drives expression in a unique pattern which may include expression in specific cell and/or tissue types within the organism (as opposed to a constitutive expression pattern in all cell types of an organism at all times).
[0069] "Wild type" or "wild-type", as used herein, refers to a plant cell, seed, plant component, plant tissue, plant organ or whole plant that has not been genetically modified or treated in an experimental sense. Wild-type cells, seed, components, tissue, organs or whole plants may be used as controls to compare levels of expression and the extent and nature of trait modification with cells, tissue or plants of the same species in which a polypeptide's expression is altered, e.g., in that it has been knocked out, overexpressed, or ectopically expressed.
[0070] With regard to gene knockouts as used herein, the term "knockout" (KO) refers to a plant or plant cell having a disruption in at least one gene in the plant or cell, where the disruption results in a reduced expression or activity of the polypeptide encoded by that gene compared to a control cell. The knockout can be the result of, for example, genomic disruptions, including transposons, tilling, and homologous recombination, antisense constructs, sense constructs, RNA silencing constructs, or RNA interference. A T-DNA insertion within a gene is an example of a genotypic alteration that may abolish expression of that gene.
[0071] "Ectopic expression" or "altered expression" in reference to a polynucleotide indicates that the pattern of expression in, e.g., a transgenic plant or plant tissue, is different from the expression pattern in a wild-type plant or a reference plant of the same species. The pattern of expression may also be compared with a reference expression pattern in a wild-type plant of the same species. For example, the polynucleotide or polypeptide is expressed in a cell or tissue type other than a cell or tissue type in which the sequence is expressed in the wild-type plant, or by expression at a time other than at the time the sequence is expressed in the wild-type plant, or by a response to different inducible agents, such as hormones or environmental signals, or at different expression levels (either higher or lower) compared with those found in a wild-type plant. The term also refers to altered expression patterns that are produced by lowering the levels of expression to below the detection level or completely abolishing expression. The resulting expression pattern can be transient or stable, constitutive or inducible. In reference to a polypeptide, the term "ectopic expression or altered expression" further may relate to altered activity levels resulting from the interactions of the polypeptides with exogenous or endogenous modulators or from interactions with factors or as a result of the chemical modification of the polypeptides.
[0072] The term "overexpression" as used herein refers to a greater expression level of a gene in a plant, plant cell or plant tissue, compared to expression of that gene in a wild-type plant, cell or tissue, at any developmental or temporal stage. Overexpression can occur when, for example, the genes encoding one or more polypeptides are under the control of a strong promoter (e.g., the cauliflower mosaic virus 35S transcription initiation region). Overexpression may also be achieved by placing a gene of interest under the control of an inducible or tissue specific promoter, or may be achieved through integration of transposons or engineered T-DNA molecules into regulatory regions of a target gene. Other means for inducing overexpression may include making targeted changes in a gene's native promoter, e.g. through elimination of negative regulatory sequences or engineering positive regulatory sequences, though the use of targeted nuclease activity (such as zinc finger nucleases or TAL effector nucleases) for genome editing. Elimination of micro-RNA binding sites in a gene's transcript may also result in overexpression of that gene. Additionally, a gene may be overexpressed by creating an artificial transcriptional activator targeted to bind specifically to its promoter sequences, comprising an engineered sequence-specific DNA binding domain such as a zinc finger protein or TAL effector protein fused to a transcriptional activation domain. Thus, overexpression may occur throughout a plant, in specific tissues of the plant, or in the presence or absence of particular environmental signals, depending on the promoter or overexpression approach used.
[0073] Overexpression may take place in plant cells normally lacking expression of polypeptides functionally equivalent or identical to the present polypeptides. Overexpression may also occur in plant cells where endogenous expression of the present polypeptides or functionally equivalent molecules normally occurs, but such normal expression is at a lower level. Overexpression thus results in a greater than normal production, or "overproduction" of the polypeptide in the plant, cell or tissue.
[0074] "Nitrogen limitation" or "nitrogen-limiting" refers to nitrogen levels that act as net limitations on primary production in terrestrial or aquatic biomes. Much of terrestrial growth, including much of crop growth, is limited by the availability of nitrogen, which can be alleviated by nitrogen input through deposition or fertilization.
[0075] "Water use efficiency", or WUE, measured as the biomass produced per unit transpiration, describes the relationship between water use and crop production. The basic physiological definition of WUE equates to the ratio of photosynthesis (A) to transpiration (T), also referred to as transpiration efficiency (Karaba et al. 2007, supra).
[0076] "Photosynthetic capacity" refers to the limits placed on photosynthesis by the physiology of the chloroplast. Specifically, regulation of light absorption and activities of enzymes in the C3 and C4 photosynthetic pathways. Increasing photosynthetic capacity is seen as an important means of increasing leaf and crop-canopy photosynthesis, and crop yield.
[0077] "Rubisco (ribulose-1,5-bisphosphate carboxylase oxygenase) activity" refers to the activation state of Rubisco, the most abundant protein in the chloroplast and a key limitation to C3 photosynthesis. Increasing Rubisco activity: by increasing the amount of Rubisco in the chloroplast; impacting any combination of specific reactions that regulate Rubisco activity; or increasing the concentration of CO2 in the chloroplast, is seen as an import means to improving C3 leaf and crop-canopy photosynthesis and crop yield.
[0078] The "capacity for RuBP (ribulose-1,5-bisphosphate) regeneration" refers to the rate at which RuBP, a key photosynthetic substrate is regenerated in the Calvin cycle. Increasing the capacity for RuBP regeneration by increasing the activity of enzymes in the regenerative phase of the Calvin cycle is seen as an important means to improving C3 leaf and crop-canopy photosynthesis and crop yield that will become progressively more important as atmospheric CO2 concentrations continue to rise.
[0079] "Leaf chlorophyll content" refers to the chlorophyll content of the leaf expressed either per unit leaf area or unit weight. Leaves absorb more light than they require for photosynthesis for large portions of the day. This absorbed energy must be dissipated if damage to the leaf is to be avoided. Consequently, decreasing leaf chlorophyll content is considered an effective means to improving photosynthetic resource-use efficiency across scales, from the leaf to the crop canopy. Decreasing leaf chlorophyll content can increase light transmission into the leaf and improve N investment between the different components of the photosynthetic apparatus. This concept can be extended to the canopy where decreased chlorophyll content in upper canopy leaves is expected to improve photosynthesis of shaded lower canopy leaves with little impact on the rate of photosynthesis of upper canopy sunlit leaves.
[0080] "Regulation of photosystem II" is a term that covers how the quantum efficiency with which electron transport is initiated as PSII responds to environment. Non-photochemical quenching from the antenna plays a key role in this regulation and is thought to constrain the efficiency of PSII operation and by extension rate of photosynthesis as leaves continually transition from high to low light. Increasing the rate of relaxation of non-photochemical quenching during the transition from high to low light is expected to increase leaf and canopy photosynthesis integrated over days to the growing season.
[0081] "Stomatal conductance" refers to a measurement of the limitation that the stomatal pore imposes on CO2 diffusion into, and H2O diffusion out of, the leaf. Decreasing stomatal conductance will decrease water loss from the leaf and crop canopy via transpiration. This will conserve soil water, delay the onset and reduce the severity of drought effects on canopy photosynthesis and other physiology.
[0082] "Yield" or "plant yield" refers to increased plant growth, increased crop growth, increased biomass, and/or increased plant product production (including grain), and is dependent to some extent on temperature, plant size, organ size, planting density, light, water and nutrient availability, and how the plant copes with various stresses, such as through temperature acclimation and water or nutrient use efficiency. Increased or improved yield may be measured as increased seed yield, increased plant product yield (plant products include, for example, plant tissue, including ground or otherwise broken-up plant tissue, and products derived from one or more types of plant tissue), or increased vegetative yield.
DESCRIPTION OF THE SPECIFIC EMBODIMENTS
Regulatory Polypeptides Modify Expression of Endogenous Genes
[0083] A regulatory polypeptide may include, but is not limited to, any polypeptide that can activate or repress transcription of a single gene or a number of genes. As one of ordinary skill in the art recognizes, regulatory polypeptides can be identified by the presence of a region or domain of structural similarity or identity to a specific consensus sequence or the presence of a specific consensus DNA-binding motif (see, for example, Riechmann et al., 2000a. supra). The plant regulatory polypeptides of the instant description belong to the MYB-(R1)R2R3 family (Shore and Sharrocks, 1995. Eur. J. Biochem. 229:1-13; Ng and Yanofsky, 2001. Nat. Rev. Genet. 2:186-195; Alvarez-Buylla et al., 2000. Proc. Natl. Acad. Sci. USA. 97:5328-5333) and are putative regulatory polypeptides.
[0084] Generally, regulatory polypeptides are involved in cell differentiation and proliferation and the regulation of growth. Accordingly, one skilled in the art would recognize that by expressing the present sequences in a plant, one may change the expression of autologous genes or induce the expression of introduced genes. By affecting the expression of similar autologous sequences in a plant that have the biological activity of the present sequences, or by introducing the present sequences into a plant, one may alter a plant's phenotype to one with improved traits related to photosynthetic resource use efficiency. The sequences of the instant description may also be used to transform a plant and introduce desirable traits not found in the wild-type cultivar or strain. Plants may then be selected for those that produce the most desirable degree of over- or under-expression of target genes of interest and coincident trait improvement.
[0085] The sequences of the present description may be from any species, particularly plant species, in a naturally occurring form or from any source whether natural, synthetic, semi-synthetic or recombinant. The sequences of the instant description may also include fragments of the present amino acid sequences. Where "amino acid sequence" is recited to refer to an amino acid sequence of a naturally occurring protein molecule, "amino acid sequence" and like terms are not meant to limit the amino acid sequence to the complete native amino acid sequence associated with the recited protein molecule.
[0086] In addition to methods for modifying a plant phenotype by employing one or more polynucleotides and polypeptides of the instant description described herein, the polynucleotides and polypeptides of the instant description have a variety of additional uses. These uses include their use in the recombinant production (i.e., expression) of proteins; as regulators of plant gene expression, as diagnostic probes for the presence of complementary or partially complementary nucleic acids (including for detection of natural coding nucleic acids); as substrates for further reactions, e.g., mutation reactions, PCR reactions, or the like; as substrates for cloning e.g., including digestion or ligation reactions; and for identifying exogenous or endogenous modulators of the regulatory polypeptides. The polynucleotide can be, e.g., genomic DNA or RNA, a transcript (such as an mRNA), a cDNA, a PCR product, a cloned DNA, a synthetic DNA or RNA, or the like. The polynucleotide can comprise a sequence in either sense or antisense orientations.
[0087] Expression of genes that encode polypeptides that modify expression of endogenous genes, polynucleotides, and proteins are well known in the art. In addition, transgenic plants comprising polynucleotides encoding regulatory polypeptides may also modify expression of endogenous genes, polynucleotides, and proteins. Examples include Peng et al., 1997. Genes Development 11: 3194-3205, and Peng et al., 1999. Nature 400: 256-261. In addition, many others have demonstrated that an Arabidopsis regulatory polypeptide expressed in an exogenous plant species elicits the same or very similar phenotypic response. See, for example, Fu et al., 2001. Plant Cell 13: 1791-1802; Nandi et al., 2000. Curr. Biol. 10: 215-218; Coupland, 1995. Nature 377: 482-483; and Weigel and Nilsson, 1995. Nature 377: 482-500).
[0088] In another example, Mandel et al., 1992b. Cell 71-133-143, and Suzuki et al., 2001. Plant J. 28: 409-418, teach that a transcription factor expressed in another plant species elicits the same or very similar phenotypic response of the endogenous sequence, as often predicted in earlier studies of Arabidopsis transcription factors in Arabidopsis (see Mandel et al., 1992a. Nature 360: 273-277; Suzuki et al., 2001. supra). Other examples include Muller et al., 2001. Plant J. 28: 169-179; Kim et al., 2001. Plant J. 25: 247-259; Kyozuka and Shimamoto, 2002. Plant Cell Physiol. 43: 130-135; Boss and Thomas, 2002. Nature, 416: 847-850; He et al., 2000. Transgenic Res. 9: 223-227; and Robson et al., 2001. Plant J. 28: 619-631.
[0089] In yet another example, Gilmour et al., 1998. Plant J. 16: 433-442 teach an Arabidopsis AP2 transcription factor, CBF1, which, when overexpressed in transgenic plants, increases plant freezing tolerance. Jaglo et al., 2001. Plant Physiol. 127: 910-917, further identified sequences in Brassica napus which encode CBF-like genes and that transcripts for these genes accumulated rapidly in response to low temperature. Transcripts encoding CBF proteins were also found to accumulate rapidly in response to low temperature in wheat, as well as in tomato. An alignment of the CBF proteins from Arabidopsis, B. napus, wheat, rye, and tomato revealed the presence of conserved consecutive amino acid residues which bracket the AP2/EREBP DNA binding domains of the proteins and distinguish them from other members of the AP2/EREBP protein family (Jaglo et al., 2001. supra).
[0090] Regulatory polypeptides mediate cellular responses and control traits through altered expression of genes containing cis-acting nucleotide sequences that are targets of the introduced regulatory polypeptide. It is well appreciated in the art that the effect of a regulatory polypeptide on cellular responses or a cellular trait is determined by the particular genes whose expression is either directly or indirectly (e.g., by a cascade of regulatory polypeptide binding events and transcriptional changes) altered by regulatory polypeptide binding. In a global analysis of transcription comparing a standard condition with one in which a regulatory polypeptide is overexpressed, the resulting transcript profile associated with regulatory polypeptide overexpression is related to the trait or cellular process controlled by that regulatory polypeptide. For example, the PAP2 gene and other genes in the Myb family have been shown to control anthocyanin biosynthesis through regulation of the expression of genes known to be involved in the anthocyanin biosynthetic pathway (Bruce et al., 2000. Plant Cell 12: 65-79; and Borevitz et al., 2000. Plant Cell 12: 2383-2393). Further, global transcript profiles have been used successfully as diagnostic tools for specific cellular states (e.g., cancerous vs. non-cancerous; Bhattacharjee et al., 2001. Proc. Natl. Acad. Sci. USA 98: 13790-13795; and Xu et al., 2001. Proc. Natl. Acad. Sci. USA 98: 15089-15094). Consequently, it is evident to one skilled in the art that similarity of transcript profile upon overexpression of different regulatory polypeptides would indicate similarity of regulatory polypeptide function.
[0091] Polypeptides and Polynucleotides of the Present Description.
[0092] The present description includes putative regulatory polypeptides, and isolated or recombinant polynucleotides encoding the polypeptides, or novel sequence variant polypeptides or polynucleotides encoding novel variants of polypeptides derived from the specific sequences provided in the Sequence Listing; the recombinant polynucleotides of the instant description may be incorporated in expression vectors for the purpose of producing transformed plants.
[0093] Because of their relatedness at the nucleotide level, the claimed sequences will typically share at least about 40% nucleotide sequence identity, or at least 45% identity, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95% or at least 96%, at least 97%, at least 98%, at least 99%, or about 100% sequence identity to one or more of the listed full-length sequences, or to a listed sequence but excluding or outside of the region(s) encoding a known consensus sequence or consensus DNA-binding site, or outside of the region(s) encoding one or all conserved domains. The degeneracy of the genetic code enables major variations in the nucleotide sequence of a polynucleotide while maintaining the amino acid sequence of the encoded protein.
[0094] Because of their relatedness at the protein level, the claimed nucleotide sequences will typically encode a polypeptide that is at least 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 90%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% or 96%, 97%, 98%, or at least 99%, or about 100% identical, in its amino acid sequence to the entire length of any of SEQ ID NOs: 2n where n=1-17.
[0095] Also provided are methods for modifying yield from a plant by modifying the mass, size or number of plant organs or seed of a plant by controlling a number of cellular processes, and for increasing a plant's photosynthetic resource use efficiency. These methods are based on the ability to alter the expression of critical regulatory molecules that may be conserved between diverse plant species. Related conserved regulatory molecules may be originally discovered in a model system such as Arabidopsis and homologous, functional molecules then discovered in other plant species. The latter may then be used to confer increased yield or photosynthetic resource use efficiency in diverse plant species.
[0096] Sequences in the Sequence Listing, derived from diverse plant species, may be ectopically expressed in overexpressor plants. The changes in the characteristic(s) or trait(s) of the plants may then be observed and found to confer increased yield and/or increased photosynthetic resource use efficiency. Therefore, the polynucleotides and polypeptides can be used to improve desirable characteristics of plants.
[0097] The polynucleotides of the instant description are also ectopically expressed in overexpressor plant cells and the changes in the expression levels of a number of genes, polynucleotides, and/or proteins of the plant cells observed. Therefore, the polynucleotides and polypeptides can be used to change expression levels of genes, polynucleotides, and/or proteins of plants or plant cells.
[0098] The data presented herein represent the results obtained in experiments with polynucleotides and polypeptides that may be expressed in plants for the purpose of increasing yield that arises from improved photosynthetic resource use efficiency.
[0099] Variants of the Disclosed Sequences.
[0100] Also within the scope of the instant description is a variant of a nucleic acid listed in the Sequence Listing, that is, one having a sequence that differs from the one of the polynucleotide sequences in the Sequence Listing, or a complementary sequence, that encodes a functionally equivalent polypeptide (i.e., a polypeptide having some degree of equivalent or similar biological activity) but differs in sequence from the sequence in the Sequence Listing, due to degeneracy in the genetic code. Included within this definition are polymorphisms that may or may not be readily detectable using a particular oligonucleotide probe of the polynucleotide encoding polypeptide, and improper or unexpected hybridization to allelic variants, with a locus other than the normal chromosomal locus for the polynucleotide sequence encoding polypeptide.
[0101] Differences between presently disclosed polypeptides and polypeptide variants are limited so that the sequences of the former and the latter are closely similar overall and, in many regions, identical. Presently disclosed polypeptide sequences and similar polypeptide variants may differ in amino acid sequence by one or more substitutions, additions, deletions, fusions and truncations, which may be present in any combination. These differences may produce silent changes and result in a functionally equivalent polypeptides. Thus, it will be readily appreciated by those of skill in the art, that any of a variety of polynucleotide sequences is capable of encoding the polypeptides and homolog polypeptides of the instant description. A polypeptide sequence variant may have "conservative" changes, wherein a substituted amino acid has similar structural or chemical properties.
[0102] Conservative substitutions include substitutions in which at least one residue in the amino acid sequence has been removed and a different residue inserted in its place. Such substitutions generally are made in accordance with the Table 1 when it is desired to maintain the activity of the protein. Table 1 shows amino acids which can be substituted for an amino acid in a protein and which are typically regarded as conservative substitutions.
TABLE-US-00001 TABLE 1 Possible conservative amino acid substitutions Amino Acid Conservative Amino Acid Conservative Residue substitutions Residue substitutions Ala Ser Leu Ile; Val Arg Lys Lys Arg; Gln Asn Gln; His Met Leu; Ile Asp Glu Phe Met; Leu; Tyr Gln Asn Pro Gly Cys Ser Ser Thr; Gly Glu Asp Thr Ser; Val Gly Pro Trp Tyr His Asn; Gln Tyr Trp; Phe Ile Leu, Val Val Ile; Leu
[0103] The polypeptides provided in the Sequence Listing have a novel activity, such as, for example, regulatory activity. Although all conservative amino acid substitutions (for example, one basic amino acid substituted for another basic amino acid) in a polypeptide will not necessarily result in the polypeptide retaining its activity, it is expected that many of these conservative mutations would result in the polypeptide retaining its activity. Most mutations, conservative or non-conservative, made to a protein but outside of a conserved domain required for function and protein activity will not affect the activity of the protein to any great extent.
[0104] Deliberate amino acid substitutions may thus be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues, as long as a significant amount of the functional or biological activity of the polypeptide is retained. For example, negatively charged amino acids may include aspartic acid and glutamic acid, positively charged amino acids may include lysine and arginine, and amino acids with uncharged polar head groups having similar hydrophilicity values may include leucine, isoleucine, and valine; glycine and alanine; asparagine and glutamine; serine and threonine; and phenylalanine and tyrosine. More rarely, a variant may have "non-conservative" changes, e.g., replacement of a glycine with a tryptophan. Similar minor variations may also include amino acid deletions or insertions, or both. Related polypeptides may comprise, for example, additions and/or deletions of one or more N-linked or O-linked glycosylation sites, or an addition and/or a deletion of one or more cysteine residues. Guidance in determining which and how many amino acid residues may be substituted, inserted or deleted without abolishing functional or biological activity may be found using computer programs well known in the art, for example, DNASTAR software (see U.S. Pat. No. 5,840,544).
[0105] Conserved Domains
[0106] Conserved domains are recurring functional and/or structural units of a protein sequence within a protein family (for example, a family of regulatory proteins), and distinct conserved domains have been used as building blocks in molecular evolution and recombined in various arrangements to make proteins of different protein families with different functions. Conserved domains often correspond to the 3-dimensional domains of proteins and contain conserved sequence patterns or motifs, which allow for their detection in polypeptide sequences with, for example, the use of a Conserved Domain Database (for example, at www.ncbi.nlm.nih.gov/cdd). The National Center for Biotechnology Information Conserved Domain Database defines conserved domains as recurring units in molecular evolution, the extents of which can be determined by sequence and structure analysis. Conserved domains contain conserved sequence patterns or motifs, which allow for their detection in polypeptide sequences (Conserved Domain Database; www.ncbi.nlm.nih.gov/Structure/cdd/cdd.shtml). A "conserved domain" or "conserved region" as used herein refers to a region in heterologous polynucleotide or polypeptide sequences where there is a relatively high degree of sequence identity between the distinct sequences. A `Myb DNA binding domain 1` is an example of a conserved domain.
[0107] Conserved domains may also be identified as regions or domains of identity to a specific consensus sequence (see, for example, Riechmann et al., 2000a. Science 290, 2105-2110; Riechmann et al., 2000b. Curr Opin Plant Biol 3: 423-434). Thus, by using alignment methods well known in the art, the conserved domains of the plant polypeptides, for example, for the first or second Myb DNA binding domain proteins may be determined. The polypeptides of Tables 2 or 3 have conserved domains specifically indicated by amino acid coordinate start and stop sites. A comparison of the regions of these polypeptides allows one of skill in the art (see, for example, Reeves and Nissen, 1990. J. Biol. Chem. 265, 8573-8582; Reeves and Nissen, 1995. Prog. Cell Cycle Res. 1: 339-349) to identify domains or conserved domains for any of the polypeptides listed or referred to in this disclosure.
[0108] Conserved domain models are generally identified with multiple sequence alignments of related proteins spanning a variety of organisms (for example, conserved domains of the disclosed sequences can be found in FIG. 2B-FIG. 2D. These alignments reveal sequence regions containing the same, or similar, patterns of amino acids. Multiple sequence alignments, three-dimensional structure and three-dimensional structure superposition of conserved domains can be used to infer sequence, structure, and functional relationships (Conserved Domain Database, supra). Since the presence of a particular conserved domain within a polypeptide is highly correlated with an evolutionarily conserved function, a conserved domain database may be used to identify the amino acids in a protein sequence that are putatively involved in functions such as binding or catalysis, as mapped from conserved domain annotations to the query sequence. For example, the presence in a protein of a first or second Myb DNA binding domain that is structurally and phylogenetically similar to one or more domains shown in Tables 2 or 3 would be a strong indicator of a related function in plants (e.g., the function of regulating and/or improving photosynthetic resource use efficiency, yield, size, biomass, and/or vigor; i.e., a polypeptide with such a domain is expected to confer altered photosynthetic resource use efficiency, yield, size, biomass, and/or vigor when its expression level is altered). Sequences herein referred to as functionally-related and/or closely-related to the sequences or domains listed in Tables 2 or 3, including polypeptides that are closely related to the polypeptides of the instant description, may have conserved domains that share at least at least nine base pairs (bp) in length and at least 72% increasing by increments of 1% to about 100% amino acid sequence identity to the sequences provided in the Sequence Listing or in Tables 2 or 3, and have similar functions in that the polypeptides of the instant description. Said polypeptides may, when their expression level is altered by suppressing their expression, knocking out their expression, or increasing their expression, confer at least one regulatory activity selected from the group consisting of increased photosynthetic resource use efficiency, greater yield, greater size, greater biomass, and/or greater vigor as compared to a control plant.
[0109] Methods using manual alignment of sequences similar or homologous to one or more polynucleotide sequences or one or more polypeptides encoded by the polynucleotide sequences may be used to identify regions of similarity and first or second Myb DNA binding domains. Such manual methods are well-known of those of skill in the art and can include, for example, comparisons of tertiary structure between a polypeptide sequence encoded by a polynucleotide that comprises a known function and a polypeptide sequence encoded by a polynucleotide sequence that has a function not yet determined. Such examples of tertiary structure may comprise predicted alpha helices, beta-sheets, amphipathic helices, leucine zipper motifs, zinc finger motifs, proline-rich regions, cysteine repeat motifs, and the like.
[0110] With respect to polynucleotides encoding presently disclosed polypeptides, a conserved domain refers to a subsequence within a polypeptide family the presence of which is correlated with at least one function exhibited by members of the polypeptide family, and which exhibits a high degree of sequence homology, such as at least 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% or 96%, 97%, 98%, 99%, or about 100% identity to a conserved domain of a polypeptide of the Sequence Listing (e.g., any of SEQ ID NOs: 61-132) or listed in Tables 2 or 3. Sequences that possess or encode for conserved domains that meet these criteria of percentage identity, and that have comparable biological and regulatory activity to the present polypeptide sequences, thus being members of the MYB19 clade polypeptides or sequences in the MYB19 clade, are described. Sequences having lesser degrees of identity but comparable biological activity are considered to be equivalents.
[0111] Orthologs and Paralogs.
[0112] Homologous sequences as described above can comprise orthologous or paralogous sequences. Several different methods are known by those of skill in the art for identifying and defining these functionally homologous sequences. General methods for identifying orthologs and paralogs, including phylogenetic methods, sequence similarity and hybridization methods, are described herein; an ortholog or paralog, including equivalogs, may be identified by one or more of the methods described below.
[0113] As described by Eisen, 1998. Genome Res. 8: 163-167, evolutionary information may be used to predict gene function. It is common for groups of genes that are homologous in sequence to have diverse, although usually related, functions. However, in many cases, the identification of homologs is not sufficient to make specific predictions because not all homologs have the same function. Thus, an initial analysis of functional relatedness based on sequence similarity alone may not provide one with a means to determine where similarity ends and functional relatedness begins. Fortunately, it is well known in the art that protein function can be classified using phylogenetic analysis of gene trees combined with the corresponding species. Functional predictions can be greatly improved by focusing on how the genes became similar in sequence (i.e., by evolutionary processes) rather than on the sequence similarity itself (Eisen, supra). In fact, many specific examples exist in which gene function has been shown to correlate well with gene phylogeny (Eisen, supra). Thus, "[t]he first step in making functional predictions is the generation of a phylogenetic tree representing the evolutionary history of the gene of interest and its homologs. Such trees are distinct from clusters and other means of characterizing sequence similarity because they are inferred by techniques that help convert patterns of similarity into evolutionary relationships . . . . After the gene tree is inferred, biologically determined functions of the various homologs are overlaid onto the tree. Finally, the structure of the tree and the relative phylogenetic positions of genes of different functions are used to trace the history of functional changes, which is then used to predict functions of [as yet] uncharacterized genes" (Eisen, supra).
[0114] Within a single plant species, gene duplication may cause two copies of a particular gene, giving rise to two or more genes with similar sequence and often similar function known as paralogs. A paralog is therefore a similar gene formed by duplication within the same species. Paralogs typically cluster together or in the same clade (a group of similar genes) when a gene family phylogeny is analyzed using programs such as CLUSTAL (Thompson et al., 1994. Nucleic Acids Res. 22: 4673-4680; Higgins et al., 1996. Methods Enzymol. 266: 383-402). Groups of similar genes can also be identified with pair-wise BLAST analysis (Feng and Doolittle, 1987. J. Mol. Evol. 25: 351-360). For example, a clade of very similar MADS domain transcription factors from Arabidopsis all share a common function in flowering time (Ratcliffe et al., 2001. Plant Physiol. 126: 122-132), and a group of very similar AP2 domain transcription factors from Arabidopsis are involved in tolerance of plants to freezing (Gilmour et al., 1998. supra). Analysis of groups of similar genes with similar function that fall within one clade can yield sub-sequences that are particular to the clade. These sub-sequences, known as consensus sequences, can not only be used to define the sequences within each clade, but define the functions of these genes; genes within a clade may contain paralogous sequences, or orthologous sequences that share the same function (see also, for example, Mount, 2001, in Bioinformatics: Sequence and Genome Analysis, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., p. 543)
[0115] Regulatory polypeptide gene sequences are conserved across diverse eukaryotic species lines (Goodrich et al., 1993. Cell 75:519-530; Lin et al., 1991. Nature 353:569-571; Sadowski et al., 1988. Nature 335: 563-564). Plants are no exception to this observation; diverse plant species possess regulatory polypeptides that have similar sequences and functions. Speciation, the production of new species from a parental species, gives rise to two or more genes with similar sequence and similar function. These genes, termed orthologs, often have an identical function within their host plants and are often interchangeable between species without losing function. Because plants have common ancestors, many genes in any plant species will have a corresponding orthologous gene in another plant species. Once a phylogenic tree for a gene family of one species has been constructed using a program such as CLUSTAL (Thompson et al., 1994. supra; Higgins et al., 1996. supra) potential orthologous sequences can be placed into the phylogenetic tree and their relationship to genes from the species of interest can be determined. Orthologous sequences can also be identified by a reciprocal BLAST strategy. Once an orthologous sequence has been identified, the function of the ortholog can be deduced from the identified function of the reference sequence.
[0116] By using a phylogenetic analysis, one skilled in the art would recognize that the ability to deduce similar functions conferred by closely-related polypeptides is predictable. This predictability has been confirmed by our own many studies in which we have found that a wide variety of polypeptides have orthologous or closely-related homologous sequences that function as does the first, closely-related reference sequence. For example, distinct regulatory polypeptides, including:
[0117] (i) AP2 family Arabidopsis G47 (found in U.S. Pat. No. 7,135,616), a phylogenetically-related sequence from soybean, and two phylogenetically-related homologs from rice all can confer greater tolerance to drought, hyperosmotic stress, or delayed flowering as compared to control plants;
[0118] (ii) CAAT family Arabidopsis G481 (found in PCT patent publication WO2004076638), and numerous phylogenetically-related sequences from eudicots and monocots can confer greater tolerance to drought-related stress as compared to control plants;
[0119] (iii) Myb-related Arabidopsis G682 (found in U.S. Pat. Nos. 7,223,904 and 7,193,129) and numerous phylogenetically-related sequences from eudicots and monocots can confer greater tolerance to heat, drought-related stress, cold, and salt as compared to control plants;
[0120] (iv) WRKY family Arabidopsis G1274 (found in U.S. Pat. No. 7,196,245) and numerous closely-related sequences from eudicots and monocots have been shown to confer increased water deprivation tolerance, and
[0121] (v) AT-hook family soy sequence G3456 (found in US patent publication 20040128712A1) and numerous phylogenetically-related sequences from eudicots and monocots, increased biomass compared to control plants when these sequences are overexpressed in plants.
[0122] The polypeptides sequences belong to distinct clades of polypeptides that include members from diverse species. In each case, most or all of the clade member sequences derived from both eudicots and monocots have been shown to confer increased yield or tolerance to one or more abiotic stresses when the sequences were overexpressed. These studies each demonstrate that evolutionarily conserved genes from diverse species are likely to function similarly (i.e., by regulating similar target sequences and controlling the same traits), and that polynucleotides from one species may be transformed into closely-related or distantly-related plant species to confer or improve traits.
[0123] Orthologs and paralogs of presently disclosed polypeptides may be cloned using compositions provided by the present description according to methods well known in the art. cDNAs can be cloned using mRNA from a plant cell or tissue that expresses one of the present sequences. Appropriate mRNA sources may be identified by interrogating Northern blots with probes designed from the present sequences, after which a library is prepared from the mRNA obtained from a positive cell or tissue. Polypeptide-encoding cDNA is then isolated using, for example, PCR, using primers designed from a presently disclosed gene sequence, or by probing with a partial or complete cDNA or with one or more sets of degenerate probes based on the disclosed sequences. The cDNA library may be used to transform plant cells. Expression of the cDNAs of interest is detected using, for example, microarrays, Northern blots, quantitative PCR, or any other technique for monitoring changes in expression. Genomic clones may be isolated using similar techniques to those.
[0124] Examples of orthologs of the Arabidopsis polypeptide sequences and their functionally similar orthologs are listed in Tables 2 or 3 and the Sequence Listing. In addition to the sequences in Tables 2 or 3 and the Sequence Listing, the claimed nucleotide sequences are phylogenetically and structurally similar to sequences listed in the Sequence Listing and can function in a plant by increasing photosynthetic resource use efficiency and/or and increasing yield, vigor, or biomass when ectopically expressed, or overexpressed, in a plant. Since a significant number of these sequences are phylogenetically and sequentially related to each other and may be shown to increase yield from a plant and/or photosynthetic resource use efficiency, one skilled in the art would predict that other similar, phylogenetically related sequences falling within the present clades of polypeptides, including MYB19 clade polypeptide sequences, would also perform similar functions when ectopically expressed.
[0125] Background Information for MYB19, and the MYB19 Clade.
[0126] A number of phylogenetic ally-related sequences have been found in other plant species. Tables 2 and 3 list a number of MYB19 clade sequences from diverse species. The tables include the SEQ ID NO: (Column 1), the species from which the sequence was derived and the Gene Identifier ("GID"; Column 2), the percent identity of the polypeptide in Column 1 to the full length MYB19 polypeptide, SEQ ID NO: 2, as determined by a BLASTp analysis, for example, with a wordlength (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (Henikoff and Henikoff, 1989. Proc. Natl. Acad. Sci. USA 89:10915; Henikoff and Henikoff, 1991. Nucleic Acids Res. 19: 6565-6572) (Column 3), the amino acid residue coordinates for the conserved first or second Myb DNA binding domains in amino acid coordinates beginning at the N-terminus, of each of the sequences (Column 4), the conserved first or second Myb DNA binding domain sequences of the respective polypeptides (Column 5); the SEQ ID NO: of each of the first or second Myb DNA binding domains (Column 6), and the percentage identity of the conserved domain in Column 5 to the conserved domain of the Arabidopsis MYB19 sequence, SEQ ID NO: 2 (as determined by a BLASTp analysis, wordlength (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix, and with the proportion of identical amino acids in parentheses; Column 7).
TABLE-US-00002 TABLE 2 Conserved `Myb DNA binding domain 1` of MYB19 and closely related sequences Col. 7 Percent Col. 4 identity Col. 3 Myb DNA Col. 6 of first Percent binding SEQ ID Myb domain Col. identity domain 1 Col. 5 NO: of in Col. 5 1 of poly- in amino Conserved Myb DNA to Myb SEQ Col. 2 peptide acid Myb DNA binding DNA binding ID Species/ in Col. 1 coordi- binding domain domain 1 of NO: Identifier to MYB19 nates domain 1 1 MYB19 2 At/MYB19 100% 17-77 WSPEEDQKLKSFILSR 61 100% (61/61) AT5G52260.1 (268/268) GHACWTTVPILAGLQ RNGKSCRLRWINYLR PGLKRGSFSEEEEET 4 At/ 60% 18-78 WSPEEDEKLRSFILSY 62 85% (52/61) AT4G25560.1 (169/280) GHSCWTTVPIKAGLQ RNGKSCRLRWINYLR PGLKRDMISAEEEET 6 Os/LOC_Os04g 48% 18-78 WSPEEDQKLRDFILRY 63 80% (49/61) 45020.1 (96/200) GHGCWSAVPVKAGL QRNGKSCRLRWINYL RPGLKHGMFSREEEET 8 Bd/Bradi5g1667 53% 18-78 WSPEEDQKLRDYIIRY 64 78% (48/61) 2.1 (102/192) GHSCWSTVPVKAGLQ RNGKSCRLRWINYLR PGLKHGMFSQEEEET 10 Zm/GRMZM2G 50% 18-78 WSPEEDQKLRDYILLH 65 77% (47/61) 170049_T01 (97/191) GHGCWSALPAKAGLQ RNGKSCRLRWINYLR PGLKHGMFSPEEEET 12 Si/Si012304m 48% 23-83 WSPEEDEKLRDFILRY 66 77% (47/61) (98/202) GHGCWSALPAKAGLQ RNGKSCRLRWINYLR PGLKHGMFSREEEET 14 Cc/clementine0. 48% 15-75 WSPEEDQRLKNYVLQ 67 77% (47/61) 9_033485m (115/237) HGHPCWSSVPINAGL QRNGKSCRLRWINYL RPGLKRGVFNMQEEE T 16 Pt/POPTR_001 50% 22-82 WSPEEDQRLRNYVLK 68 77% (47/61) 5s13190.1 (109/217) HGHGCWSSVPINAGL QRNGKSCRLRWINYL RPGLKRGTFSAQEEET 18 Eg/EUCGR.K0 49% 18-78 WSPEEDQKLRNYVLK 69 76% (46/60) 0250.1 (107/217) HGHGCWSSVPINTGL QRNGKSCRLRWINYL RPGLKRGMFTMEEEEI 20 Eg/EUCGR.K0 48% 18-78 WSPEEDQRLRNYILNH 70 75% (45/60) 0251.1 (110/226) GHGYWSSVPINTGLQ RNGKSCRLRWINYLR PGLKRGMFTLEEEEI 22 Pt/POPTR_001 48% 18-78 WSPEEDQRLGSYVFQ 71 75% (46/61) 2s13260.1 (109/223) HGHGCWSSVPINAGL QRTGKSCRLRWINYL RPGLKRGAFSTDEEET 24 Gm/Glyma16g3 48% 18-78 WSPEEDNKLRNHIIKH 72 75% (46/61) 1280.1 (116/238) GHGCWSSVPIKAGLQ RNGKSCRLRWINYLR PGLKRGVFSKHEEDT 26 Gm/Glyma09g2 49% 18-78 WSPEEDNKLRNHIIKH 73 73% (45/61) 5590.1 (103/209) GHGCWSSVPIKAGLQ RNGKSCRLRWINYLR PGLKRGVFSKHEKDT 28 Sl/Solyc03g025 40% 52-112 WSPDEDDRLKNYMIK 74 73% (44/60) 870.2.1 (115/283) HGHGCWSSVPINAGL QRNGKSCRLRWINYL RPGLKRGAFSLEEEDI 30 Vv/GSVIVT010 42% 22-82 WSPEEDARLRNYVLK 75 72% (44/61) 28984001 (115/272) YGLGCWSSVPVNAGL QRNGKSCRLRWINYL RPGLKRGMFTIEEEET 32 Eg/EUCGR.A0 51% 20-80 WSPDEDQRLRNYIHK 76 70% (44/61) 2796.1 (112/217) HGYSCWSSVPINAGL QRNGKSCRLRWINYL RPGLKRGAFTVQEEET 34 At/AT3G48920. 51%) 19-79 WSPEEDEKLRSHVLK 77 69% (41-59) 1 (99/191) YGHGCWSTIPLQAGL QRNGKSCRLRWVNYL RPGLKKSLFTKQEETI
TABLE-US-00003 TABLE 3 Conserved second Myb DNA binding domains of MYB19 and closely related sequences Col. 7 Percent Col. 4 identity Col. 3 Myb DNA of second Percent binding Col. 6 Myb domain Col. identity domain 2 Col. 5 SEQ ID in Col. 5 1 of poly- in amino Conserved NO: of to Myb SEQ Col. 2 peptide acid Myb DNA second DNA binding ID Species/ in Col. 1 coordi- binding Myb domain 2 of NO: Identifier to MYB19 nates domain 2 domain MYB19 2 At/MYB19 100% 70-112 FSEEEEETILTLHSSL 95 100% (43/43) AT5G52260.1 (268/268) GNKWSRIAKYLPGRT DNEIKNYWHSYL 4 At/ 60% 68-110 ISAEEEETILTFHSSLG 96 88% (37/42) AT4G25560.1 (169/280) NKWSQIAKFLPGRTD NEIKNYWHSHL 6 Os/LOC_Os04g 48% 71-113 FSREEEETVMNLHAT 97 72% (31/43) 45020.1 (96/200) MGNKWSQIARHLPG RTDNEVKNYWNSYL 8 Bd/ 53% 71-113 FSQEEEETVMSLHAT 98 76% (33/43) Bradi5g16672.1 (102/192) LGNKWSRIAQHLPGR TDNEVKNYWNSYL 10 Zm/GRMZM2 50% 71-113 FSPEEEETVMSLHAT 99 76% (33/43) G170049_T01 (97/191) LGNKWSRIARHLPGR TDNEVKNYWNSYL 12 Si/Si012304m 48% 71-113 FSREEEETVMSLHAK 100 74% (32/43) (98/202) LGNKWSQIARHLPGR TDNEVKNYWNSYL 14 Cc/clementine0. 48% 75-117 FNMQEEETILTVHRL 101 76% (33/43) 9_033485m (115/237) LGNKWSQIAQHLPGR TDNEIKNYWHSHL 16 Pt/POPTR_001 50% 75-117 FSAQEEETILALHHM 102 79% (34/43) 5s13190.1 (109/217) LGNKWSQIAQHLPGR TDNEIKNHWHSYL 18 Eg/EUCGR.K0 49% 71-113 FTMEEEEIIFSLHHLIG 103 74% (32/43) 0250.1 (107/217) NKWSQIAKHLPGRTD NEIKNHWHSYL 20 Eg/EUCGR.K0 48% 71-113 FTLEEEEIILSLHRLIG 104 76% (33/43) 0251.1 (110/226) NKWSQIAKHLPGRTD NEIKNHWHSYL 22 Pt/POPTR_001 48% 105-147 FSTDEEETILTLHRML 105 81% (35/43) 2s13260.1 (109/223) GNKWSQIAQHLPGRT DNEIKNHWHSYL 24 Gm/Glyma16g3 48% 71-113 FSKHEEDTIMVLHHM 106 76% (33/43) 1280.1 (116/238) LGNKWSQIAQHLPGR TDNEIKNYWHSYL 26 Gm/Glyma09g2 49% 71-113 FSKHEKDTIMALHH 107 72% (31/43) 5590.1 (103/209) MLGNKWSQIAQHLP GRTDNEVKNYWHSY L 28 Sl/Solyc03g025 40% 72-114 FSLEEEDIILTLHAMF 108 76% (33/43) 870.2.1 (115/283) GNKWSQIAQQLPGRT DNEIKNHWHSYL 30 Vv/GSVIVT01 42% 73-115 FTIEEEETIMALHRLL 109 74% (32/43) 028984001 (115/272) GNKWSQIAQNFPGRT DNEIKNYWHSCL 32 Eg/EUCGR.A0 51% 71-113 FTVQEEETILNLHHLL 110 76% (33/43) 2796.1 (112/217) GNKWSQIAQHLPGRT DNEIKNHWHSYL 34 At/AT3G48920. 51%) 76-118 FTKQEETILLSLHSML 111 72% (31/43) 1 (99/191) GNKWSQISKFLPGRT DNEIKNYWHSNL Species abbreviations for Tables 2 and 3: At- Arabidopsis thaliana; Bd- Brachypodium distachyon; Cc- Citrus x clementina; Eg- Eucalyptus grandis; Gm- Glycine max; Os- Oryza sativa; Pt- Populus trichocarpa; Si- Setaria italica; Sl- Solanum lycopersicum; Vv- Vitis vinifera; Zm- Zea mays
[0127] Sequences that are functionally-related and/or closely-related to the polypeptides in Tables 2 and 3 may be created artificially, semi-synthetically, or may occur naturally by having descended from the same ancestral sequence as the disclosed MYB19-related sequences, where the polypeptides have the function of conferring increased photosynthetic resource use efficiency to plants. These "functionally-related and/or closely-related" MYB19 clade polypeptides generally contain the consensus sequence of the Myb DNA binding domain 1 of SEQ ID NO: 129:
WSPX1EDxxLxxxX2xxxGxxxWX3x X2PxxxGLQRxGKSCRLRWX2NYLRPGLKxxxxxxxE; where x represents any amino acid;
X1 is D or E;
X2 is I, V, L or M;
[0128] and X3 represents S or T; as provided in FIG. 2B-2C.
[0129] Other highly conserved residues found in the Myb DNA binding domain 2 of MYB19 clade members, as shown in FIG. 2C-2D and SEQ ID NO: 130:
ExxxX1xxxHxxxGNKWSxIX2xxxPGRTDNEX1KNxWxSxL where x represents any amino acid;
X1 is I, V, L or M; and
[0130] X2 represents A or S.
[0131] There is also a small motif that is present in MYB19 clade member proteins, identifiable as SEQ ID NO: 160 and that can be located spanning FIGS. 2E-2F:
PxFxX1W
[0132] where x represents any amino acid; and
X1 is D or E.
[0133] The presence of one or more of these consensus sequences and/or these amino acid residues is correlated with conferring of improved or increased photosynthetic resource use efficiency to a plant when the expression level of the polypeptide is altered in a plant by being reduced, knocked-out, or overexpressed. A MYB19 clade polypeptide sequence that is "functionally-related and/or closely-related" to the listed full length protein sequences or domains provided in Tables 2 or 3 may also have at least 40%, 42%, 48%, 49%, 50%, 51%, 53%, 60%, or about 100% amino acid identity to SEQ ID NO: 2 or to SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, and/or at least 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or about 100% amino acid identity to the first Myb DNA binding domain of SEQ ID NO: 2, or to a listed first Myb DNA binding domain or to SEQ ID NOs: 61-77, and/or 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or about 100% amino acid identity to a listed second Myb DNA binding domain or to the second Myb DNA binding domain of SEQ ID NO: 2 or SEQ ID NOs: 95-111, or to an amino acid sequence having at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95% or at least 96%, at least 97%, at least 98%, at least 99%, or about 100% sequence identity to SEQ ID NOs: 129-132. The presence of the disclosed conserved first Myb DNA binding domains and/or second Myb DNA binding domains in the polypeptide sequence (for example, SEQ ID NO: 61-77 or 95-111), is correlated with the conferring of improved or increased photosynthetic resource use efficiency to a plant when the expression level of the polypeptide is altered in a plant by being reduced, knocked-out, or overexpressed. All of the sequences that adhere to these functional and sequential relationships are herein referred to as "MYB19 clade polypeptides" or "MYB19 clade polypeptides", or which fall within the "MYB19 clade" or "G1309 clade" exemplified in the tree in FIG. 1 as those polypeptides bounded by LOC_Os04g45020.1 and Solyc03g025870.2.1 (indicated by the box around these sequences).
Examples of Methods for Identifying Identity, Similarity, Homology and Relatedness
[0134] Percent identity can be determined electronically, e.g., by using the MEGALIGN program (DNASTAR, Inc. Madison, Wis.) or the Accelrys. The MEGALIGN program can create alignments between two or more sequences according to different methods, for example, the clustal method (see, for example, Higgins and Sharp, 1988. Gene 73: 237-244). The clustal algorithm groups sequences into clusters by examining the distances between all pairs. The clusters are aligned pairwise and then in groups. Other alignment algorithms or programs may be used for preparing alignments and/or determining percentage identities, including Accelrys Gene, FASTA, BLAST, or ENTREZ, FASTA and BLAST, some of which may also be used to calculate percent similarity. Accelrys Gene is available from Accelrys, Inc., San Diego, Calif. Other programs are available as a part of the GCG sequence analysis package (University of Wisconsin, Madison, Wis.), and can be used with or without default settings. ENTREZ is available through the National Center for Biotechnology Information. In one embodiment, the percent identity of two sequences can be determined by the GCG program with a gap weight of 1, e.g., each amino acid gap is weighted as if it were a single amino acid or nucleotide mismatch between the two sequences (see U.S. Pat. No. 6,262,333).
[0135] Software for performing BLAST analyses is publicly available, e.g., through the National Center for Biotechnology Information (see internet website at www.ncbi.nlm.nih.gov/). This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul, 1990. J. Mol. Biol. 215: 403-410; Altschul, 1993. J. Mol. Evol. 36: 290-300). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, a cutoff of 100, M=5, N=-4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff, 1989. supra; Henikoff and Henikoff, 1991. supra). Unless otherwise indicated for comparisons of predicted polynucleotides, "sequence identity" refers to the % sequence identity generated from a tBLASTx using the NCBI version of the algorithm at the default settings using gapped alignments with the filter "off" (see, for example, internet website at www.ncbi.nlm nih gov).
[0136] Other techniques for alignment are described by Doolittle, ed., 1996. Methods in Enzymology, vol. 266: "Computer Methods for Macromolecular Sequence Analysis" Academic Press, Inc., San Diego, Calif., USA. Preferably, an alignment program that permits gaps in the sequence is utilized to align the sequences. The Smith-Waterman is one type of algorithm that permits gaps in sequence alignments (see Shpaer, 1997. Methods Mol. Biol. 70: 173-187). Also, the GAP program using the Needleman and Wunsch alignment method can be utilized to align sequences. An alternative search strategy uses MPSRCH software, which runs on a MASPAR computer. MPSRCH uses a Smith-Waterman algorithm to score sequences on a massively parallel computer. This approach improves ability to pick up distantly related matches, and is especially tolerant of small gaps and nucleotide sequence errors. Nucleic acid-encoded amino acid sequences can be used to search both protein and DNA databases.
[0137] The percentage similarity between two polypeptide sequences, e.g., sequence A and sequence B, is calculated by dividing the length of sequence A, minus the number of gap residues in sequence A, minus the number of gap residues in sequence B, into the sum of the residue matches between sequence A and sequence B, times one hundred. Gaps of low or of no similarity between the two amino acid sequences are not included in determining percentage similarity. Percent identity between polynucleotide sequences can also be counted or calculated by other methods known in the art, e.g., the Jotun Hein method (see, for example, Hein, 1990. Methods Enzymol. 183: 626-645). Identity between sequences can also be determined by other methods known in the art, e.g., by varying hybridization conditions (see US Patent Application No. 20010010913).
[0138] The percent identity between two polypeptide sequences can also be determined using Accelrys Gene v2.5, 2006. with default parameters: Pairwise Matrix: GONNET; Align Speed: Slow; Open Gap Penalty: 10.000; Extended Gap Penalty: 0.100; Multiple Matrix: GONNET; Multiple Open Gap Penalty: 10.000; Multiple Extended Gap Penalty: 0.05; Delay Divergent: 30; Gap Separation Distance: 8; End Gap Separation: false; Residue Specific Penalties: false; Hydrophilic Penalties: false; Hydrophilic Residues: GPSNDQEKR. The default parameters for determining percent identity between two polynucleotide sequences using Accelrys Gene are: Align Speed: Slow; Open Gap Penalty: 10.000; Extended Gap Penalty: 5.000; Multiple Open Gap Penalty: 10.000; Multiple Extended Gap Penalty: 5.000; Delay Divergent: 40; Transition: Weighted.
[0139] In addition, one or more polynucleotide sequences or one or more polypeptides encoded by the polynucleotide sequences may be used to search against a BLOCKS (Bairoch et al., 1997. Nucleic Acids Res. 25: 217-221), PFAM, and other databases which contain previously identified and annotated motifs, sequences and gene functions. Methods that search for primary sequence patterns with secondary structure gap penalties (Smith et al., 1992. Protein Engineering 5: 35-51) as well as algorithms such as Basic Local Alignment Search Tool (BLAST; Altschul, 1990. supra; Altschul et al., 1993. supra), BLOCKS (Henikoff and Henikoff, 1991 supra), Hidden Markov Models (HMM; Eddy, 1996. Curr. Opin. Str. Biol. 6: 361-365; Sonnhammer et al., 1997. Proteins 28: 405-420), and the like, can be used to manipulate and analyze polynucleotide and polypeptide sequences encoded by polynucleotides. These databases, algorithms and other methods are well known in the art and are described in Ausubel et al., 1997. Short Protocols in Molecular Biology, John Wiley & Sons, New York, N.Y., unit 7.7, and in Meyers, 1995. Molecular Biology and Biotechnology, Wiley VCH, New York, N.Y., p 856-853.
[0140] Thus, the instant description provides methods for identifying a sequence similar or paralogous or orthologous or homologous to one or more polynucleotides as noted herein, or one or more target polypeptides encoded by the polynucleotides, or otherwise noted herein and may include linking or associating a given plant phenotype or gene function with a sequence. In the methods, a sequence database is provided (locally or across an internet or intranet) and a query is made against the sequence database using the relevant sequences herein and associated plant phenotypes or gene functions.
[0141] A further method for identifying or confirming that specific homologous sequences control the same function is by comparison of the transcript profile(s) obtained upon overexpression or knockout of two or more related polypeptides. Since transcript profiles are diagnostic for specific cellular states, one skilled in the art will appreciate that genes that have a highly similar transcript profile (e.g., with greater than 50% regulated transcripts in common, or with greater than 70% regulated transcripts in common, or with greater than 90% regulated transcripts in common) will have highly similar functions. Fowler and Thomashow, 2002. Plant Cell 14, 1675-1690, have shown that three paralogous AP2 family genes (CBF1, CBF2 and CBF3) are induced upon cold treatment, each of which can condition improved freezing tolerance, and all have highly similar transcript profiles. Once a polypeptide has been shown to provide a specific function, its transcript profile becomes a diagnostic tool to determine whether paralogs or orthologs have the same function.
[0142] Identifying Polynucleotides or Nucleic Acids by Hybridization.
[0143] Polynucleotides homologous to the sequences illustrated in the Sequence Listing and tables can be identified, e.g., by hybridization to each other under stringent or under highly stringent conditions. Stringency is influenced by a variety of factors, including temperature, salt concentration and composition, organic and non-organic additives, solvents, etc. present in both the hybridization and wash solutions and incubations, and the number of washes, as described in more detail in the references cited below (e.g., Sambrook et al., 1989. supra; Berger and Kimmel, eds., 1987. Methods Enzymol. 152: 507-511; Anderson and Young, 1985. "Quantitative Filter Hybridisation", In: Hames and Higgins, ed., Nucleic Acid Hybridisation, A Practical Approach. Oxford, IRL Press, 73-111), each of which are incorporated herein by reference. Conditions that are highly stringent, and means for achieving them, are also well known in the art and described in, for example, Sambrook et al., 1989. supra; Berger and Kimmel, eds., 1987. Meth. Enzymol. 152:467-469; and Anderson and Young, 1985. supra.
[0144] Also provided in the instant description are polynucleotide sequences that are capable of hybridizing to the claimed polynucleotide sequences, including any of the polynucleotides within the Sequence Listing, and fragments thereof under various conditions of stringency (see, for example, Wahl and Berger, 1987. Methods Enzymol. 152: 399-407; Berger and Kimmel, ed., 1987. Methods Enzymol. 152:507-511). In addition to the nucleotide sequences listed in the Sequence Listing, full length cDNA, orthologs, and paralogs of the present nucleotide sequences may be identified and isolated using well-known methods. The cDNA libraries, orthologs, and paralogs of the present nucleotide sequences may be screened using hybridization methods to determine their utility as hybridization target or amplification probes.
[0145] Stability of DNA duplexes is affected by such factors as base composition, length, and degree of base pair mismatch. Hybridization conditions may be adjusted to allow DNAs of different sequence relatedness to hybridize. The melting temperature (Tm) is defined as the temperature when 50% of the duplex molecules have dissociated into their constituent single strands. The melting temperature of a perfectly matched duplex, where the hybridization buffer contains formamide as a denaturing agent, may be estimated by the following equations:
[0146] (I) DNA-DNA:
Tm(° C.)=81.5+16.6(log [Na+])+0.41(% G+C)-0.62(% formamide)-500/L
[0147] (II) DNA-RNA:
Tm(° C.)=79.8+18.5(log [Na+])+0.58(% G+C)+0.12(% G+C)2-0.5(% formamide)-820/L
[0148] (III) RNA-RNA:
Tm(° C.)=79.8+18.5(log [Na+])+0.58(% G+C)+0.12(% G+C)2-0.35(% formamide)-820/L
[0149] where L is the length of the duplex formed, [Na+] is the molar concentration of the sodium ion in the hybridization or washing solution, and % G+C is the percentage of (guanine+cytosine) bases in the hybrid. For imperfectly matched hybrids, approximately 1° C. is required to reduce the melting temperature for each 1% mismatch.
[0150] Hybridization experiments are generally conducted in a buffer of pH between 6.8 to 7.4, although the rate of hybridization is nearly independent of pH at ionic strengths likely to be used in the hybridization buffer (Anderson and Young, 1985. supra). In addition, one or more of the following may be used to reduce non-specific hybridization: sonicated salmon sperm DNA or another non-complementary DNA, bovine serum albumin, sodium pyrophosphate, sodium dodecylsulfate (SDS), polyvinyl-pyrrolidone, ficoll and Denhardt's solution. Dextran sulfate and polyethylene glycol 6000 act to exclude DNA from solution, thus raising the effective probe DNA concentration and the hybridization signal within a given unit of time. In some instances, conditions of even greater stringency may be desirable or required to reduce non-specific and/or background hybridization. These conditions may be created with the use of higher temperature, lower ionic strength and higher concentration of a denaturing agent such as formamide.
[0151] Stringency conditions can be adjusted to screen for moderately similar fragments such as homologous sequences from distantly related organisms, or to highly similar fragments such as genes that duplicate functional enzymes from closely related organisms. The stringency can be adjusted either during the hybridization step or in the post-hybridization washes. Salt concentration, formamide concentration, hybridization temperature and probe lengths are variables that can be used to alter stringency (as described by the formula above). As a general guideline, high stringency is typically performed at Tm-5° C. to Tm-20° C., moderate stringency at Tm-20° C. to Tm-35° C. and low stringency at Tm-35° C. to Tm-50° C. for duplex >150 base pairs. Hybridization may be performed at low to moderate stringency (25-50° C. below Tm), followed by post-hybridization washes at increasing stringencies. Maximum rates of hybridization in solution are determined empirically to occur at Tm-25° C. for DNA-DNA duplex and Tm-15° C. for RNA-DNA duplex. Optionally, the degree of dissociation may be assessed after each wash step to determine the need for subsequent, higher stringency wash steps.
[0152] High stringency conditions may be used to select for nucleic acid sequences with high degrees of identity to the disclosed sequences. An example of stringent hybridization conditions obtained in a filter-based method such as a Southern or Northern blot for hybridization of complementary nucleic acids that have more than 100 complementary residues is about 5° C. to 20° C. lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. Conditions used for hybridization may include about 0.02 M to about 0.15 M sodium chloride, about 0.5% to about 5% casein, about 0.02% SDS or about 0.1% N-laurylsarcosine, about 0.001 M to about 0.03 M sodium citrate, at hybridization temperatures between about 50° C. and about 70° C. More preferably, high stringency conditions are about 0.02 M sodium chloride, about 0.5% casein, about 0.02% SDS, about 0.001 M sodium citrate, at a temperature of about 50° C. Nucleic acid molecules that hybridize under stringent conditions will typically hybridize to a probe based on either the entire DNA molecule or selected portions, e.g., to a unique subsequence, of the DNA.
[0153] Stringent salt concentration will ordinarily be less than about 750 mM NaCl and 75 mM trisodium citrate. Increasingly stringent conditions may be obtained with less than about 500 mM NaCl and 50 mM trisodium citrate, to even greater stringency with less than about 250 mM NaCl and 25 mM trisodium citrate. Low stringency hybridization can be obtained in the absence of organic solvent, e.g., formamide, whereas high stringency hybridization may be obtained in the presence of at least about 35% formamide, and more preferably at least about 50% formamide. Stringent temperature conditions will ordinarily include temperatures of at least about 30° C., more preferably of at least about 37° C., and most preferably of at least about 42° C. with formamide present. Varying additional parameters, such as hybridization time, the concentration of detergent, e.g., sodium dodecyl sulfate (SDS) and ionic strength, are well known to those skilled in the art. Various levels of stringency are accomplished by combining these various conditions as needed.
[0154] The washing steps that follow hybridization may also vary in stringency; the post-hybridization wash steps primarily determine hybridization specificity, with the most critical factors being temperature and the ionic strength of the final wash solution. Wash stringency can be increased by decreasing salt concentration or by increasing temperature. Stringent salt concentration for the wash steps will preferably be less than about 30 mM NaCl and 3 mM trisodium citrate, and most preferably less than about 15 mM NaCl and 1.5 mM trisodium citrate.
[0155] Thus, high stringency hybridization and wash conditions that may be used to bind and remove polynucleotides with less than the desired homology to the nucleic acid sequences or their complements that encode the present polypeptides include, for example:
[0156] 6×SSC at 65° C.;
[0157] 50% formamide, 4×SSC at 42° C.; or
[0158] 0.5×SSC, 0.1% SDS at 65° C.;
[0159] with, for example, two wash steps of 10-30 minutes each. Useful variations on these conditions will be readily apparent to those skilled in the art.
[0160] A person of skill in the art would not expect substantial variation among polynucleotide species provided with the present description because the highly stringent conditions set forth in the above formulae yield structurally similar polynucleotides.
[0161] If desired, one may employ wash steps of even greater stringency, including about 0.2×SSC, 0.1% SDS at 65° C. and washing twice, each wash step being about 30 minutes, or about 0.1×SSC, 0.1% SDS at 65° C. and washing twice for 30 minutes. The temperature for the wash solutions will ordinarily be at least about 25° C., and for greater stringency at least about 42° C. Hybridization stringency may be increased further by using the same conditions as in the hybridization steps, with the wash temperature raised about 3° C. to about 5° C., and stringency may be increased even further by using the same conditions except the wash temperature is raised about 6° C. to about 9° C. For identification of less closely related homologs, wash steps may be performed at a lower temperature, e.g., 50° C.
[0162] An example of a low stringency wash step employs a solution and conditions of at least 25° C. in 30 mM NaCl, 3 mM trisodium citrate, and 0.1% SDS over 30 minutes. Greater stringency may be obtained at 42° C. in 15 mM NaCl, with 1.5 mM trisodium citrate, and 0.1% SDS over 30 minutes. Even higher stringency wash conditions are obtained at 65° C.-68° C. in a solution of 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. Wash procedures will generally employ at least two final wash steps. Additional variations on these conditions will be readily apparent to those skilled in the art (see, for example, US Patent Application No. 20010010913).
[0163] Stringency conditions can be selected such that an oligonucleotide that is perfectly complementary to the coding oligonucleotide hybridizes to the coding oligonucleotide with at least about a 5-10× higher signal to noise ratio than the ratio for hybridization of the perfectly complementary oligonucleotide to a nucleic acid encoding a polypeptide known as of the filing date of the application. It may be desirable to select conditions for a particular assay such that a higher signal to noise ratio, that is, about 15× or more, is obtained. Accordingly, a subject nucleic acid will hybridize to a unique coding oligonucleotide with at least a 2× or greater signal to noise ratio as compared to hybridization of the coding oligonucleotide to a nucleic acid encoding known polypeptide. The particular signal will depend on the label used in the relevant assay, e.g., a fluorescent label, a colorimetric label, a radioactive label, or the like. Labeled hybridization or PCR probes for detecting related polynucleotide sequences may be produced by oligolabeling, nick translation, end-labeling, or PCR amplification using a labeled nucleotide.
[0164] The present description also provides polynucleotide sequences that are capable of hybridizing to the claimed polynucleotide sequences, including any of the polynucleotides within the Sequence Listing, and fragments thereof under various conditions of stringency (see, for example, Wahl and Berger, 1987, supra, pages 399-407; and Kimmel, 1987. Meth. Enzymol. 152, 507-511). In addition to the nucleotide sequences in the Sequence Listing, full length cDNA, orthologs, and paralogs of the present nucleotide sequences may be identified and isolated using well-known methods. The cDNA libraries, orthologs, and paralogs of the present nucleotide sequences may be screened using hybridization methods to determine their utility as hybridization target or amplification probes.
EXAMPLES
[0165] It is to be understood that this description is not limited to the particular devices, machines, materials and methods described. Although particular embodiments are described, equivalent embodiments may be used to practice the claims.
[0166] The specification, now being generally described, will be more readily understood by reference to the following examples, which are included merely for purposes of illustration of certain aspects and embodiments of the present description and are not intended to limit the claims or description. It will be recognized by one of skill in the art that a polypeptide that is associated with a particular first trait may also be associated with at least one other, unrelated and inherent second trait which was not predicted by the first trait.
Example I
Plant Genotypes and Vector and Cloning Information
[0167] A variety of constructs may be used to modulate the activity of regulatory polypeptides (RPs), and to test the activity of orthologs and paralogs in transgenic plant material. This platform provides the material for all subsequent analysis.
[0168] An individual plant "genotype" refers to a set of plant lines containing a particular construct or knockout (for example, this might be 35S lines for a given gene sequence (GID, Gene Identifier) being tested, 35S lines for a paralog or ortholog of that gene sequence, lines for an RNAi construct, lines for a GAL4 fusion construct, or lines in which expression of the gene sequence is driven from a particular promoter that enhances expression in particular cell, tissue or condition). For a given genotype arising from a particular transformed construct, multiple independent transgenic lines may be examined for morphological and physiological phenotypes. Each individual "line" (also sometimes known as an "event") refers to the progeny plant or plants deriving from the stable integration of the transgene(s), carried within the T-DNA borders contained within a transformation construct, into a specific location or locations within the genome of the original transformed cell. It is well known in the art that different lines deriving from transformation with a given transgene may exhibit different levels of expression of that transgene due to so called "position effects" of the surrounding chromatin at the locus of integration in the genome, and therefore it is necessary to examine multiple lines containing each construct of interest.
(1) Overexpression/Tissue-Enhanced/Conditional Expression
[0169] Expression of a given regulatory protein from a particular promoter, for example a photosynthetic tissue-enhanced promoter (e.g., a green tissue- or leaf-enhanced promoter), is achieved either by a direct-promoter fusion construct in which that regulatory protein is cloned directly behind the promoter of interest or by a two component system.
[0170] The Two-Component Expression System.
[0171] For the two-component system, two separate constructs are used: Promoter::LexA-GAL4TA and opLexA::RP. The first of these (Promoter::LexA-GAL4TA) comprises a desired promoter cloned in front of a LexA DNA binding domain fused to a GAL4 activation domain. The construct vector backbone (pMEN48, also known as P5375) also carries a kanamycin resistance marker, along with an opLexA::GFP (green fluorescent protein) reporter. Transgenic lines are obtained containing this first component, and a line is selected that shows reproducible expression of the reporter gene in the desired pattern through a number of generations. A homozygous population is established for that line, and the population is supertransformed with the second construct (opLexA::RP) carrying the regulatory protein of interest cloned behind a LexA operator site. This second construct vector backbone (pMEN53, also known as P5381) also contains a sulfonamide resistance marker.
[0172] Conditional Expression.
[0173] Various promoters can be used to overexpress disclosed polypeptides in plants to confer improved photosynthetic resource use efficiency. However, in some cases, there may be limitations in the use of various proteins that confer increased photosynthetic resource use efficiency when the proteins are overexpressed. Negative side effects associated with constitutive overexpression such as small size, delayed growth, increased disease sensitivity, and development and alteration in flowering time are not uncommon. A number of stress-inducible promoters can be used promote protein expression during the periods of stress, and therefore may be used to induce overexpression of polypeptides that can confer improved stress tolerance when they are needed without the adverse developmental or morphological effects that may be associated with their constitutive overexpression.
[0174] Promoters that drive protein expression in response to stress can be used to regulate the expression of the disclosed polypeptides to confer photosynthetic resource use efficiency to plants. The promoter may regulate expression of a disclosed polypeptide to an effective level in a photosynthetic tissue. Effective level in this regard refers to an expression level that confers greater photosynthetic resource use efficiency in the transgenic plant relative to the control plant that, for example, does not comprise a recombinant polynucleotide that encodes the disclosed polypeptide. Optionally, the promoter does not regulate protein expression in a constitutive manner.
[0175] Such promoters include, but are not limited to, the sequences located in the promoter regions of At5g52310 (RD29A), At5g52300, AT1G16850, At3g46230, AT1G52690, At2g37870, AT5G43840, At5g66780, At3g17520, and At4g09600.
[0176] In addition, promoters with expression specific to or enhanced in particular cells or tissue types may be used to express a given regulatory protein only in these cells or tissues. Examples of such promoter types include but are not limited to promoters expressed in green tissue, guard cell, epidermis, whole root, root hairs, vasculature, apical meristems, and developing leaves.
[0177] Table 4 lists a number of photosynthetic tissue-enhanced promoters, specifically, mesophyll tissue-enhanced promoters from rice, that may be used to regulate expression of polynucleotides and polypeptides found in the Sequence Listing and structurally and functionally-related sequences. Promoters that may be used to drive expression of polynucleotides and polypeptides found in the Sequence Listing and structurally and functionally-related sequences included, but are not limited to, promoter sequences listed in Table 4.
TABLE-US-00004 TABLE 4 Rice Genes with Photosynthetic Tissue-Enhanced Promoters Rice Gene Identifier of Photosynthetic SEQ ID NO: Tissue-Enhanced Promoter 136 Os02g09720 137 Os05g34510 138 Os11g08230 139 Os01g64390 140 Os06g15760 141 Os12g37560 142 Os03g17420 143 Os04g51000 144 Os01g01960 145 Os05g04990 146 Os02g44970 147 Os01g25530 148 Os03g30650 149 Os01g64910 150 Os07g26810 151 Os07g26820 152 Os09g11220 153 Os04g21800 154 Os10g23840 155 Os08g13850 156 Os12g42980 157 Os03g29280 158 Os03g20650 159 Os06g43920
[0178] Tissue-enhanced promoters that may be used to drive expression of polynucleotides and polypeptides found in the Sequence Listing and structurally and functionally-related sequences have also been described in US patent application U520110179520A1, incorporated herein by reference. Such promoters include, but are not limited to, Arabidopsis sequences located in the promoter regions of AT1G08465, AT1G10155, AT1G14190, AT1G24130, AT1G24735, AT1G29270, AT1G30950, AT1G31310, AT1G37140, AT1G49320, AT1G49475, AT1G52100, AT1G60540, AT1G60630, AT1G64625, AT1G65150, AT1G68480, AT1G68780, AT1G69180, AT1G77145, AT1G80580, AT2G03500, AT2G17950, AT2G19910, AT2G27250, AT2G33880, AT2G39850, AT3G02500, AT3G12750, AT3G15170, AT3G16340, AT3G27920, AT3G30340, AT3G42670, AT3G44970, AT3G49950, AT3G50870, AT3G54990, AT3G59270, AT4G00180, AT4G00480, AT4G12450, AT4G14819, AT4G31610, AT4G31615, AT4G31620, AT4G31805, AT4G31877, AT4G36060, AT4G36470, AT4G36850, AT4G37970, AT5G03840, AT5G12330, AT5G14070, AT5G16410, AT5G20740, AT5G27690, AT5G35770, AT5G39330, AT5G42655, AT5G53210, AT5G56530, AT5G58780, AT5G61070, and AT5G6491.
[0179] In addition to the sequences provided in the Sequence Listing or in this Example, a promoter region may include a fragment of the promoter sequences provided in the Sequence Listing or in this Example, or a complement thereof, wherein the promoter sequence, or the fragment thereof, or the complement thereof, regulates expression of a polypeptide in a plant cell, for example, in response to a biotic or abiotic stress, or in a manner that is enhanced or preferred in certain plant tissues.
(2) Knock-Out/Knock-Down
[0180] In some cases, lines mutated in a given regulatory protein may be analyzed. Where available, T-DNA insertion lines in a given gene are isolated and characterized. In cases where a T-DNA insertion line is unavailable, an RNA interference (RNAi) strategy is sometimes used.
Example II
Transformation Methods
[0181] Transformation of Monocots.
[0182] Cereal plants including corn, wheat, rice, sorghum, barley, or other monocots may be transformed with the present polynucleotide sequences, including monocot or eudicot-derived sequences such as those presented in the present Tables, cloned into a vector such as pGA643 and containing a kanamycin-resistance marker, and expressed constitutively under, for example, the CaMV35S or COR15 promoters, or with tissue-enhanced or inducible promoters. The expression vectors may be one found in the Sequence Listing, or any other suitable expression vector may be similarly used. For example, pMEN020 may be modified to replace the NptII coding region with the BAR gene of Streptomyces hygroscopicus that confers resistance to phosphinothricin. The KpnI and BgIII sites of the Bar gene are removed by site-directed mutagenesis with silent codon changes.
[0183] The cloning vector may be introduced into a variety of cereal plants by means well known in the art including direct DNA transfer or Agrobacterium tumefaciens-mediated transformation. The latter approach may be accomplished by a variety of means, including, for example, that of U.S. Pat. No. 5,591,616, in which monocotyledon callus is transformed by contacting dedifferentiating tissue with the Agrobacterium containing the cloning vector.
[0184] The sample tissues are immersed in a suspension of 3×10-9 cells of Agrobacterium containing the cloning vector for 3-10 minutes. The callus material is cultured on solid medium at 25° C. in the dark for several days. The calli grown on this medium are transferred to a Regeneration Medium. Transfers are continued every two to three weeks (two or three times) until shoots develop. Shoots are then transferred to Shoot-Elongation Medium every 2-3 weeks. Healthy looking shoots are transferred to Rooting Medium and after roots have developed, the plants are placed into moist potting soil.
[0185] The transformed plants are then analyzed for the presence of the NPTII gene/kanamycin resistance by ELISA, using the ELISA NPTII kit from SPrime-3Prime Inc. (Boulder, Colo.).
[0186] It is also routine to use other methods to produce transgenic plants of most cereal crops (Vasil, 1994. Plant Mol. Biol. 25: 925-937) such as corn, wheat, rice, sorghum (Cassas et al., 1993. Proc. Natl. Acad. Sci. USA 90: 11212-11216), and barley (Wan and Lemeaux, 1994. Plant Physiol. 104: 37-48). DNA transfer methods such as the microprojectile method can be used for corn (Fromm et al., 1990. Bio/Technol. 8: 833-839; Gordon-Kamm et al., 1990. Plant Cell 2: 603-618; Ishida, 1990. Nature Biotechnol. 14:745-750), wheat (Vasil et al., 1992. Bio/Technol. 10:667-674; Vasil et al., 1993. Bio/Technol. 11:1553-1558; Weeks et al., 1993. Plant Physiol. 102:1077-1084), and rice (Christou, 1991. Bio/Technol. 9:957-962; Hiei et al., 1994. Plant J. 6:271-282; Aldemita and Hodges, 1996. Planta 199: 612-617; and Hiei et al., 1997. Plant Mol. Biol. 35:205-218). For most cereal plants, embryogenic cells derived from immature scutellum tissues are the preferred cellular targets for transformation (Hiei et al., 1997. supra; Vasil, 1994. supra). For transforming corn embryogenic cells derived from immature scutellar tissue using microprojectile bombardment, the A188XB73 genotype is the preferred genotype (Fromm et al., 1990. Bio/Technol. 8: 833-839; Gordon-Kamm et al., 1990. supra). After microprojectile bombardment the tissues are selected on phosphinothricin to identify the transgenic embryogenic cells (Gordon-Kamm et al., 1990. supra). Transgenic plants from transformed host plant cells may be regenerated by standard corn regeneration techniques (Fromm et al., 1990. Bio/Technol. 8: 833-839; Gordon-Kamm et al., 1990. supra).
[0187] Transformation of Dicots.
[0188] Crop species that overexpress polypeptides of the instant description may produce plants with increased photosynthetic resource use efficiency and/or yield. Thus, polynucleotide sequences listed in the Sequence Listing recombined into, for example, one of the expression vectors of the instant description, or another suitable expression vector, may be transformed into a plant for the purpose of modifying plant traits for the purpose of improving yield, quality, and/or photosynthetic resource use efficiency. The expression vector may contain a constitutive, tissue-enhanced or inducible promoter operably linked to the polynucleotide. The cloning vector may be introduced into a variety of plants by means well known in the art such as, for example, direct DNA transfer or Agrobacterium tumefaciens-mediated transformation. It is now routine to produce transgenic plants using most eudicot plants (see Weissbach and Weissbach, 1989. Methods for Plant Molecular Biology, Academic Press; Gelvin et al., 1990. Plant Molecular Biology Manual, Kluwer Academic Publishers; Herrera-Estrella et al., 1983. Nature 303: 209; Bevan, 1984. Nucleic Acids Res. 12: 8711-8721; and Klee, 1985. Bio/Technology 3: 637-642). Methods for analysis of traits are routine in the art and examples are disclosed above.
[0189] Numerous protocols for the transformation of tomato and soy plants have been previously described, and are well known in the art. Gruber et al., in Glick and Thompson, 1993. Methods in Plant Molecular Biology and Biotechnology. eds., CRC Press, Inc., Boca Raton, describe several expression vectors and culture methods that may be used for cell or tissue transformation and subsequent regeneration. For soybean transformation, methods are described by Miki et al., 1993. in Methods in Plant Molecular Biology and Biotechnology, p. 67-88, Glick and Thompson, eds., CRC Press, Inc., Boca Raton; and U.S. Pat. No. 5,563,055, (Townsend and Thomas), issued Oct. 8, 1996.
[0190] There are a substantial number of alternatives to Agrobacterium-mediated transformation protocols, other methods for the purpose of transferring exogenous genes into soybeans or tomatoes. One such method is microprojectile-mediated transformation, in which DNA on the surface of microprojectile particles is driven into plant tissues with a biolistic device (see, for example, Sanford et al., 1987. Part. Sci. Technol. 5:27-37; Sanford, 1993. Methods Enzymol. 217: 483-509; Christou et al., 1992. Plant. J. 2: 275-281; Klein et al., 1987. Nature 327: 70-73; U.S. Pat. No. 5,015,580 (Christou et al), issued May 14, 1991; and U.S. Pat. No. 5,322,783 (Tomes et al.), issued Jun. 21, 1994).
[0191] Alternatively, sonication methods (see, for example, Zhang et al., 1991. Bio/Technology 9: 996-997); direct uptake of DNA into protoplasts using CaCl2 precipitation, polyvinyl alcohol or poly-L-ornithine (see, for example, Hain et al., 1985. Mol. Gen. Genet. 199: 161-168; Draper et al., 1982. Plant Cell Physiol. 23: 451-458); liposome or spheroplast fusion (see, for example, Deshayes et al., 1985. EMBO J., 4: 2731-2737; Christou et al., 1987. Proc. Natl. Acad. Sci. USA 84: 3962-3966); and electroporation of protoplasts and whole cells and tissues (see, for example, Donn et al. (1990. in Abstracts of VIIth International Congress on Plant Cell and Tissue Culture IAPTC, A2-38: 53; D'Halluin et al., 1992. Plant Cell 4: 1495-1505; and Spencer et al., 1994. Plant Mol. Biol. 24: 51-61) have been used to introduce foreign DNA and expression vectors into plants.
[0192] After a plant or plant cell is transformed (and the transformed host plant cell then regenerated into a plant), the transformed plant may propagated vegetatively or it may be crossed with itself or a plant from the same line, a non-transformed or wild-type plant, or another transformed plant from a different transgenic line of plants. Crossing provides the advantages of producing new and often stable transgenic varieties. Genes and the traits they confer that have been introduced into a tomato or soybean line may be moved into distinct line of plants using traditional backcrossing techniques well known in the art. Transformation of tomato plants may be conducted using the protocols of Koornneef et al, 1986. In Tomato Biotechnology: Alan R. Liss, Inc., 169-178, and in U.S. Pat. No. 6,613,962, the latter method described in brief here. Eight day old cotyledon explants are precultured for 24 hours in Petri dishes containing a feeder layer of Petunia hybrida suspension cells plated on MS medium with 2% (w/v) sucrose and 0.8% agar supplemented with 10 μM α-naphthalene acetic acid and 4.4 μM 6-benzylaminopurine. The explants are then infected with a diluted overnight culture of Agrobacterium tumefaciens containing an expression vector comprising a polynucleotide of the instant description for 5-10 minutes, blotted dry on sterile filter paper and cocultured for 48 hours on the original feeder layer plates. Culture conditions are as described above. Overnight cultures of Agrobacterium tumefaciens are diluted in liquid MS medium with 2% (w/v/) sucrose, pH 5.7) to an OD600 of 0.8.
[0193] Following cocultivation, the cotyledon explants are transferred to Petri dishes with selective medium comprising MS medium with 4.56 μM zeatin, 67.3 μM vancomycin, 418.9 μM cefotaxime and 171.6 μM kanamycin sulfate, and cultured under the culture conditions described above. The explants are subcultured every three weeks onto fresh medium. Emerging shoots are dissected from the underlying callus and transferred to glass jars with selective medium without zeatin to form roots. The formation of roots in a kanamycin sulfate-containing medium is a positive indication of a successful transformation.
[0194] Transformation of soybean plants may be conducted using the methods found in, for example, U.S. Pat. No. 5,563,055 (Townsend et al., issued Oct. 8, 1996), described in brief here. In this method soybean seed is surface sterilized by exposure to chlorine gas evolved in a glass bell jar. Seeds are germinated by plating on 1/10 strength agar solidified medium without plant growth regulators and culturing at 28° C. with a 16 hour day length. After three or four days, seed may be prepared for cocultivation. The seedcoat is removed and the elongating radicle removed 3-4 mm below the cotyledons.
[0195] Eucalyptus is now considered an important crop that is grown for example to provide feedstocks for the pulp and paper and biofuel markets. This species is also amenable to transformation as described in PCT patent publication WO/2005/032241.
[0196] Crambe has been recognized as a high potential oilseed crop that may be grown for the production of high value oils. An efficient method for transformation of this species has been described in PCT patent publication WO 2009/067398 A1.
[0197] Overnight cultures of Agrobacterium tumefaciens harboring the expression vector comprising a polynucleotide of the instant description are grown to log phase, pooled, and concentrated by centrifugation. Inoculations are conducted in batches such that each plate of seed was treated with a newly resuspended pellet of Agrobacterium. The pellets are resuspended in 20 ml inoculation medium. The inoculum is poured into a Petri dish containing prepared seed and the cotyledonary nodes are macerated with a surgical blade. After 30 minutes the explants are transferred to plates of the same medium that has been solidified. Explants are embedded with the adaxial side up and level with the surface of the medium and cultured at 22° C. for three days under white fluorescent light. These plants may then be regenerated according to methods well established in the art, such as by moving the explants after three days to a liquid counter-selection medium (see U.S. Pat. No. 5,563,055).
[0198] The explants may then be picked, embedded and cultured in solidified selection medium. After one month on selective media transformed tissue becomes visible as green sectors of regenerating tissue against a background of bleached, less healthy tissue. Explants with green sectors are transferred to an elongation medium. Culture is continued on this medium with transfers to fresh plates every two weeks. When shoots are 0.5 cm in length they may be excised at the base and placed in a rooting medium.
Experimental Methods; Transformation of Arabidopsis
[0199] Transformation of Arabidopsis is performed by an Agrobacterium-mediated protocol based on the method of Bechtold and Pelletier, 1998. Unless otherwise specified, all experimental work is performed using the Columbia ecotype.
[0200] Plant Preparation.
[0201] Arabidopsis seeds are gas sterilized and sown on plates with media containing 80% MS with vitamins, 0.3% sucrose and 1% Bacto agar. The plates are placed at 4° in the dark for the days then transferred to 24 hour light at 22° for 7 days. After 7 days the seedlings are transplanted to soil, placing individual seedlings in each pot. The primary bolts are cut off a week before transformation to break apical dominance and encourage auxiliary shoots to form. Transformation is typically performed at 4-5 weeks after sowing.
[0202] Bacterial Culture Preparation.
[0203] Agrobacterium stocks are inoculated from single colony plates or from glycerol stocks and grown with the appropriate antibiotics until saturation. On the morning of transformation, the saturated cultures are centrifuged and bacterial pellets are re-suspended in Infiltration Media (0.5×MS, 1× Gamborg's Vitamins, 5% sucrose, 200 μl/L Silwet L77) until an A600 reading of 0.8 is reached.
[0204] Transformation and Harvest of Transgenic Seeds.
[0205] The Agrobacterium solution is poured into dipping containers. All flower buds and rosette leaves of the plants are immersed in this solution for 30 seconds. The plants are laid on their side and wrapped to keep the humidity high. The plants are kept this way overnight at 22° C. and then the pots are turned upright, unwrapped, and moved to the growth racks. In most cases, the transformation process is repeated one week later to increase transformation efficiency.
[0206] The plants are maintained on the growth rack under 24-hour light until seeds are ready to be harvested. Seeds are harvested when 80% of the siliques of the transformed plants are ripe (approximately five weeks after the initial transformation). This seed is deemed T0 seed, since it is obtained from the T0 generation, and is later plated on selection plates (either kanamycin or sulfonamide). Resistant plants that are identified on such selection plates comprise the T1 generation, from which transgenic seed comprising an expression vector of interest may be derived.
Example III
Primary Screening Materials and Methods
[0207] Plant Growth Conditions
[0208] Seeds from Arabidopsis lines are chlorine gas sterilized using a standard protocol and spread onto plates containing a sucrose based media augmented with vitamins (80% MS+Vit, 1% sucrose, 0.65% PhytoBlend Agar (Caisson Laboratories, Inc., North Logan, Utah) and appropriate kanamycin or sulfonamide concentrations where selection is required. Seeds are stratified in the dark on plates, at 4° C. for 3 days then moved to a walk-in growth chamber (Conviron MTW120, Conviron Controlled Environments Ltd, Winnipeg, Manitoba, Canada) running at a 10 hour photoperiod at a photosynthetic photon flux of approximately 200 μmol m-2 s-1 at plant height and a photoperiod/night temperature regime of 22° C./19° C. After seven days of light exposure seedlings are transplanted into 164 ml volume pots containing autoclaved ProMix® soil. All pots are returned to the same growth-chamber where they are stood in water and covered with a lid for the first seven days. This protocol keeps the soil moist during this period. Seven days after transplanting lids are removed and a watering and nutrition regime begun. All plants receive water three times a week, and a weekly a fertilizer treatment (80% Peter's NPK fertilizer).
[0209] Primary Screening
[0210] Between 35 and 38 days after being transferred to light on plates, and after between 28 and 31 days growth in soil, a suite of leaf-physiological parameters are measured using an infrared gas analyzer (LI-6400XT, LI-COR® Biosciences, Lincoln, NB, USA) integrated with `a fluorimeter that measures fluorescence from Chlorophyll A (LI-6400-40, LI-COR Biosciences). The growth conditions used, and plant age and leaf selection criteria for measurement are designed to maximize the chance that the leaves sampled fill the 2 cm2 leaf chamber of the gas-exchange system and that plants show no visible signs of having transitioned to reproductive growth.
[0211] Screening High-Light Leaf Physiology at Two Air Temperatures
[0212] Leaf physiology is screened after plants have been acclimated to high light (700 μmol photons m-2 s-1) under LED light banks emitting visible light (400-700 nm, Photon Systems Instruments, Brno, Czech Republic), for 40 minutes. Other than the change in light level, the atmospheric environment is the same as that in which the plants have been grown, and the LI-6400 leaf chamber is set to reflect this, being set to deliver a photosynthetic photon flux of 700 μmol photons m-2 s-1 and operate at an air temperature of 22° C. Forty minutes acclimation to a photosynthetic photon flux of 700 μmol photons m-2 s-1 has repeatedly been shown to be sufficient to achieve a steady-state rate of light-saturated photosynthesis and stomatal conductance in control plants. Gas exchange and fluorescence data are logged simultaneously two minutes after the leaf has been closed in the chamber. Two minutes is found to be long enough for the leaf chamber CO2 and H2O concentrations to stabilize after closing a new leaf inside, and thereby minimizing leaf physiological adjustment to small differences between the growth environment and the LI-6400 chamber. Screening at the growth air temperature of 22° C. is begun one hour into the photoperiod and is typically completed in two hours. After being screened at 22° C., plants are returned to growth-light levels prior to being screened again at 35° C. later in the photoperiod. The higher-temperature screening begins six hours into the photoperiod and measurements are made after the rosettes have been acclimated to the same high light dose as described above, but this time in a controlled environment with an air temperature set to 35° C. Measurements are again made in a leaf chamber set to match the warmer air temperature and logged using the protocol described above for the 22° C. measurements. Data generated at both 22° C. and 35° C. are used to calculate: rates of CO2 assimilation by photosynthesis (A, μmol CO2 m-2 s-1); rates of H2O loss through transpiration (Tr, mmol H2O m-2 s-1); the conductance to CO2 and H2O movement between the leaf and air through the stomatal pore (gs, mol. m-2 s-1); the sub-stomatal CO2 concentration (Ci, μmol CO2 mol-1); transpiration efficiency, the instantaneous ratio of photosynthesis to transpiration, (TE=A/Tr (μmol CO2 mmol H2O m-2 s-1)); the rate of electron flow through photosystem two (ETR μmol e-m-2 s-1). Derivation of the parameters described above followed established published protocols (Long & Bernacchi, 2003. J. Exp. Botany; 54:2393-24)
[0213] Leaves from up to 10 replicate plants are screened for a given line of interest. Data generated from these lines are compared with that from an empty vector control line planted at the same time, and grown within the same flats, as the lines being screened.
[0214] For control lines, data are collected not only at an atmospheric CO2 concentration of 400 μmol CO2 mol-1, but also after stepwise changes in CO2 concentration to 350, 300, 450 and 500 μmol CO2 mol-1. These measurements underlay screening for more complex physiological traits of: 1) photosynthetic capacity; 2) regulation of photosystem two (PSII) operation; and 3) non-photosynthetic metabolism.
[0215] Screening Photosynthetic Capacity
[0216] Under most conditions, the rate of light-saturated photosynthesis in a C3 leaf is a product of the biochemical capacity of the Calvin cycle and the transfer conductance of CO2 concentration to the sites of carboxylation (Farquhar et al., 1980. Planta:149, 78-90). Plotting the rate of photosynthesis against an estimate of the sub-stomatal CO2 concentration (Ci) provides a means to identify changes in photosynthetic capacity of the Calvin cycle independent of changes in stomatal conductance, a key component of the total transfer conductance to CO2 of the leaf. Consequently, for lines being screened, rates of photosynthesis are plotted against a regression plot of A vs. Ci generated for the control lines over a range of atmospheric CO2 concentration, as described above. This technique enables visual confirmation of changes in photosynthetic capacity in lines of interest.
[0217] Screening Regulation of Photosystem Two (PSII) Operation
[0218] During acclimation to high light, the efficiency with which photosystem PSII operates will reach a steady state regulated largely by the feedback between non-photochemical quenching in the antenna and the metabolic demand for energy produced in the chloroplast (Genty et al., 1989. Biochim. Biophys. Acta 990:87-92; Baker et al., 2007. Plant Cell Environ. 30:1107-1125). This understanding is used in this screen to identify lines in which the limitation that non-photochemical quenching exerts on the efficiency with which photosystem II operates, is decreased. Lower levels of non-photochemical quenching will result in a higher efficiency of photosynthesis over a range of light levels, but importantly higher rates of photosynthesis at low light where light-use efficiency is important. Increasing rates of photosynthesis as leaves in crop canopies transition from high to low light is a process thought relevant to increasing crop-canopy photosynthesis (Zhu et al., 2010. Plant Biol. 61:235-261). In keeping with the A/Ci analysis described above, a regression of the operating efficiency of PSII against non-photochemical quenching is generated for the control line from data collected over a range of atmospheric CO2 concentration to provide a reference against which data for lines of interest can be visually compared.
[0219] Screening for Non-Photosynthetic Metabolism
[0220] Measurement of the ratio of the rate of electron flow through PSII (ETR) to the rate of photosynthesis (A) is used to screen for changes in non-photosynthetic metabolism. This screen is based upon the understanding that the transport of four μmol of electrons from PSII to photosystem one PSI will supply the NADPH and ATP required to fix one μmol of CO2 in the Calvin cycle. For a C3 leaf operating in an atmosphere with 21% oxygen, the ratio of electron flow to photosynthesis should be higher than four, reflecting photorespiratory and other metabolism. However, because the rate of photorespiration in a C3 leaf is dependent upon the concentration of CO2 at the active site of Rubisco, a regression of the ratio of electron flow to photosynthesis, generated over the range of CO2 concentrations described above, provides the reference regression against which lines being screened can be compared to controls. Changes in the ratio of ETR to A, when observed at the same C, as the control line, could indicate changes in the specificity of the Rubisco active site for O2 relative to CO2 and or other metabolic sinks which would be expected to have important implications for crop productivity and/or stress tolerance.
[0221] Surrogate Screening for Growth-Light Physiology
[0222] Rosette biomass: the dry weight of whole Arabidopsis rosettes is measured after being dried down at 80° C. for 24 hours, a time found to be sufficient to reach constant weight. Samples are taken after 35-38 days growth, and used as an assay of above-ground productivity at growth light. Typically, five replicate rosettes are sampled per Arabidopsis line being screened.
[0223] Rosette chemical and isotopic C and N analysis: after weighing, the five rosettes sampled for each line screened are pooled together and ground to a fine powder. The pooled sample generated is sub-sampled and approximately 4 μg samples are prepared for analysis.
[0224] Chlorophyll content index (CCI): measurements of light transmission through the leaf are made for plants being screened using a chlorophyll content meter (CCM-200, Apogee Instruments, Logan, Utah, USA). The first is made within the first hour of the photoperiod prior to any acclimation to high light on leaves of plants samples for rosette analysis. The second is made later in the photoperiod on leaves of plants that had undergone the high-temperature screening.
[0225] Light absorption: measurements of CCI are used as a surrogate for leaf light absorption, based upon a known relationship between the two. The estimates of light absorption by the leaf, required to construct this relationship, were made by placing the leaf on top of a quantum sensor (LI-190, LI-COR Biosciences) with both the leaf and quantum sensor then pressed firmly up to the foam gasket underneath the LI-6400 light source. This procedure provides an estimate of the transmission of a known light flux through the leaf and is used to estimate the fraction of light absorbed by the leaf.
Example IV
Experimental Results
[0226] This Example provides experimental observations for transgenic plants overexpressing AtMYB19 related polypeptides in plate-based assays and results observed for improved photosynthetic resource use efficiency.
[0227] Photosynthetic rate was increased in six of nine independent lines screened at growth temperature (22° C.) and seven of nine lines for measurements made after acclimation to high temperature. For measurements made at air temperatures of 22° C. and 35° C.; photosynthesis was increased by 16% at 22° C. and 17% at 35° C., when averaged across the lines that displayed increased photosynthesis. This provided evidence that the increase in photosynthesis is conferred over a wide range of air temperatures observed in Arabidopsis plants overexpressing AtMYB19. Leaf and crop-canopy photosynthesis is known to be related to final crop yield and improving photosynthesis is widely considered to be a relevant pathway to increasing crop yield. In a C3 plant, photosynthesis at high-light can be limited by the biochemical capacity for photosynthesis, indicated as photosynthetic capacity in Tables 5 and 6, or the supply of CO2 into the chloroplast, of which stomatal conductance, which regulates the transfer of CO2 into the leaf through stoma, is a principle component. Both the capacity for photosynthesis and stomatal conductance were increased in Arabidopsis plants overexpressing AtMYB19 assayed at both temperatures. Photosynthetic capacity was increased in five lines at 22° C. and in three at 35° C. Focused secondary assays on select lines, enabled the biochemical limitations to photosynthesis that underlay photosynthetic capacity, to be investigated. For measurements made at 22° C., the biochemical basis for the increase in photosynthetic capacity was an increase in the plant's capacity to regenerate RuBP, a key substrate for photosynthesis, for four lines (figure *). Two of these four lines also displayed evidence of an increase in the activity of Rubisco (figure *). For measurements made at 35° C., three lines displayed an increase in the capacity to regenerate RuBP. Stomatal conductance was increased by 32% at 22° C. and 37% at 35° C., when averaged across the AtMYB19 overexpression lines that displayed increased photosynthesis. The extent to which photosynthesis is increased as a consequence of improvements in photosynthetic capacity and stomatal conductance has important implications. For example, increasing stomatal conductance will increase the supply of CO2 into the leaf, however this will increase photosynthesis to a greater extent in a C3 plant than a C4 plant, where chloroplast CO2 concentrations are typically maintained at close to saturating levels for photosynthesis. Increasing stomatal conductance will increase transpiration from the leaf, typically to a greater extent than photosynthesis is stimulated. This combination of traits may be more appropriate for crops growing on acreages where soil-water availability is seldom limiting yield. Conversely, an increase in photosynthetic capacity could increase photosynthetic rate without increasing stomatal conductance and water loss, and would be expected to increase crop yield over broad acres. For transgenic plants overexpressing AtMYB19 related polypeptides, the increase in photosynthetic rate was the result of increases in both photosynthetic capacity and stomatal conductance. Consequently transpiration efficiency, often used synonymously with WUE and expressed as unit carbon uptake via photosynthesis per unit water lost via transpiration, was typically not decreased across lines and temperatures.
[0228] All experimental observations of greater photosynthetic resource use efficiency were made by comparison to control plants (e.g., plants that did not comprise a recombinant construct encoding a AtMYB19-related polypeptide or overexpress a AtMYB19 clade or phylogenetically-related regulatory protein).
[0229] Tables 5 and 6 present the indicators of photosynthetic resource use efficiency observed in Arabidopsis plants overexpressing AtMYB19 in experiments conducted to date. The data presented in Table 5 were collected on plants at their normal growth temperature of 22° C. For lines with increased photosynthetic capacity, RuBP indicates that the capacity to increase RuBP was increased and Rubisco indicates that Rubisco activity was increased.
TABLE-US-00005 TABLE 5 Photosynthetic resource use efficiency measurements in plants with altered expression of MYB19 clade polypeptides at a growth temperature of 22° C. Polypeptide SEQ Photosynthetic Stomatal sequence/ ID Rate Conductance Photosynthetic Line NO Driver Target 22° C. 22° C. Capacity MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased No effect Line 1 GAL4_opLexA::GFP (20%) (32%) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased Increased Line 2 GAL4_opLexA::GFP (15%) (28%) (Rubisco and RuBP) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased Increased Line 3 GAL4_opLexA::GFP (10%) (35%) (Rubisco and RuBP) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 No effect No effect No effect Line 4 GAL4_opLexA::GFP MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased Increased Line 5 GAL4_opLexA::GFP (26%) (27%) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased Increased Line 6 GAL4_opLexA::GFP (13%) (30%) RuBP MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased Increased Line 7 GAL4_opLexA::GFP (10%) (41%) RuBP MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 No effect No effect No effect Line 8 GAL4_opLexA::GFP
[0230] The data presented in Table 6 were collected on plants acclimated to an air temperature of 35° C. For lines with increased photosynthetic capacity, RuBP indicates that the capacity to increase RuBP was increased and Rubisco indicates that Rubisco activity was increased.
TABLE-US-00006 TABLE 6 Photosynthetic resource use efficiency measurements in plants with altered expression of MYB19 clade polypeptides at a growth temperature of 35° C. Polypeptide Seq Photosynthetic Stomatal sequence/ ID Rate Conductance Photosynthetic Line No Driver Target 22° C. 22° C. Capacity MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased No effect Line 1 GAL4_opLexA::GFP (22%) (49%) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased Increased Line 2 GAL4_opLexA::GFP (14%) (43%) (RuBP) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased Increased Line 3 GAL4_opLexA::GFP (15%) (23%) (RuBP) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased No effect Line 4 GAL4_opLexA::GFP (26%) (39%) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased No effect Line 5 GAL4_opLexA::GFP (22%) (37%) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased No effect Line 6 GAL4_opLexA::GFP (19%) (61%) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 Increased Increased Increased Line 7 GAL4_opLexA::GFP (13%) (28%) (RuBP) MYB19/ 2 35S::m35S::oEnh:LexA: opLexA::G1309 No effect Increased No effect Line 8 GAL4_opLexA::GFP (17%)
[0231] The results presented in Tables 5 and 6 were determined after screening nine independent transgenic events. Multiple lines were screened in replicate independent experiments.
[0232] The present disclosure thus describes how the transformation of plants, which may include monocots and/or dicots, with a MYB19 clade polypeptide can confer to the transformed plants greater photosynthetic resource use efficiency than the level of photosynthetic resource use efficiency exhibited by control plants. In one embodiment, expression of MYB19 is driven by a constitutive promoter. In another embodiment, expression of MYB19 is driven by a promoter with enhanced activity in a tissue capable of photosynthesis (also referred to herein as a "photosynthetic promoter" or a "photosynthetic tissue-enhanced promoter") such as a leaf tissue or other green tissue. Examples of photosynthetic tissue-enhanced promoters include for example, an RBCS3 promoter (SEQ ID NO: 133), an RBCS4 promoter (SEQ ID NO: 134) or others such as the At4g01060 promoter (SEQ ID NO: 135), the latter regulating expression in guard cells. Other photosynthetic tissue-enhanced promoters have been taught by Bassett et al., 2007. BMC Biotechnol. 7: 47, specifically incorporated herein by reference in its entirety. Other photosynthetic tissue-enhanced promoters of interest include those from the maize aldolase gene FDA (U.S. Patent Publication No. U520040216189, specifically incorporated herein by reference in its entirety, and the aldolase and pyruvate orthophosphate dikinase (PPDK) (Taniguchi et al., 2000. Plant Cell Physiol. 41:42-48, specifically incorporated herein by reference in its entirety. Other tissue enhanced promoters or inducible promoters are also envisioned that may be used to regulate expression of MYB19 clade member polypeptides and improve photosynthetic resource use efficiency in a variety of plants.
Example V
Utilities of MYB19 Clade Sequences for Improving Photosynthetic Resource Use Efficiency, Yield or Biomass
[0233] The improved photosynthetic resource use efficiency conferred by increasing the expression level of a MYB19 clade polypeptide sequence may contribute to increased yield of commercially available plants. For plants for which biomass is the product of interest, increasing the expression level of MYB19 clade of polypeptide sequences may increase yield, photosynthetic resource use efficiency, vigor, growth rate, and/or biomass of the plants. Thus, it is thus expected that these sequences will improve yield and/or photosynthetic resource use efficiency in non-Arabidopsis plants relative to control plants. This yield improvement may result in yield increases in crop or non-Arabidopsis plants including, but not limited to, wheat, Setaria, corn (maize), rice, barley; rye; millet; sorghum; sugarcane, miscane, turfgrass, Miscanthus, switchgrass; soybean, cotton, rape, oilseed rape including canola, Eucalyptus or poplar, such as at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30% or greater yield relative to the yield that may be obtained with control plants.
[0234] It is expected that the same methods may be applied to identify other useful and valuable sequences that are functionally-related and/or closely-related to the listed sequences or domains provided in Tables 2 or 3, and the sequences may be derived from a diverse range of species. Because of morphological, physiological and photosynthetic resource use efficiency similarities that may occur among MYB19-related sequences, the MYB19 clade sequences are expected to increase yield, plant growth, vigor, size, biomass, and/or increase photosynthetic resource use efficiency to a variety of crop plants, ornamental plants, and woody plants used in the food, ornamental, paper, pulp, lumber or other industries.
Example VI
Expression and Analysis of Increased Yield or Photosynthetic Resource Use Efficiency in Non-Arabidopsis or Crop Species
[0235] Northern blot analysis, RT-PCR or microarray analysis of the regenerated, transformed plants may be used to show expression of a polypeptide or the instant description and related genes that are capable of inducing improved photosynthetic resource use efficiency, and/or larger size.
[0236] After a eudicot plant, monocot plant or plant cell has been transformed (and the latter plant host cell regenerated into a plant) and shown to have greater photosynthetic resource use efficiency, and/or greater size, vigor, biomass, and/or produce greater yield relative to a control plant, the transformed monocot plant may be crossed with itself or a plant from the same line, a non-transformed or wild-type monocot plant, or another transformed monocot plant from a different transgenic line of plants.
[0237] The function of one or more specific polypeptides of the instant description has been analyzed and may be further characterized and incorporated into crop plants. The ectopic overexpression of one or more of MYB19 clade polypeptide sequences may be regulated using constitutive, inducible, or tissue-enhanced regulatory elements. Genes that have been examined have been shown to modify plant traits including increasing yield and/or photosynthetic resource use efficiency. It is expected that newly discovered polynucleotide and polypeptide sequences closely related, as determined by the disclosed hybridization or identity analyses, to polynucleotide and polypeptide sequences found in the Sequence Listing can also confer alteration of traits in a similar manner to the sequences found in the Sequence Listing, when transformed into any of a considerable variety of plants of different species, and including dicots and monocots. The polynucleotide and polypeptide sequences derived from monocots (e.g., the rice sequences) may be used to transform both monocot and dicot plants, and those derived from dicots (e.g., the Arabidopsis and soy genes) may be used to transform either group, although it is expected that some of these sequences will function best if the gene is transformed into a plant from the same group as that from which the sequence is derived.
[0238] As an example of a first step to determine photosynthetic resource use efficiency, seeds of these transgenic plants may be grown as described above or methods known in the art. Disclosed sequences may be identified that, when ectopically expressed, or overexpressed, in plants, the expression of said sequences result in one or more characteristics that lead to greater photosynthetic resource use efficiency.
These characteristics include:
[0239] (a) increased photosynthetic capacity, measured as an increase in the rate of light-saturated photosynthesis of at least 10% when compared to the rate of light-saturated photosynthesis of a control leaf at the same leaf-internal CO2 concentration. Optionally, measurements are made after 40 minutes of acclimation to a light intensity that is saturating for photosynthesis;
[0240] (b) Increased photosynthetic rate, measured as an increase in the rate of light-saturated photosynthesis of at least 10%. Optionally, measurements are made after 40 minutes of acclimation to a light intensity known to be saturating for photosynthesis
[0241] (c) a decrease in the chlorophyll content of the leaf of at least 10%, observed in the absence of a decrease in photosynthetic capacity;
[0242] (d) a decrease in the percentage of the leaf dry weight that is nitrogen of at least 0.5%, observed in the absence of a decrease in photosynthetic capacity or increase in dry weight;
[0243] (e) increased transpiration efficiency, measured as an increase in the rate of light-saturated photosynthesis relative to water loss via transpiration from the leaf, of at least 10%; optionally, measurements are made after 40 minutes of acclimation to a light intensity of 700 μmol PAR m-2 s-1;
[0244] (f) an increase in the resistance to water vapor diffusion out of the leaf that is exerted by the stomata, measured as a decrease in stomatal conductance to H2O loss from the leaf of at least 10%; optionally, measurements were are after 40 minutes of acclimation to a light intensity of 700 μmol PAR m-2 s-1;
[0245] (g) a decrease in the relative limitation that non-photochemical quenching exerts on the operation of PSII. The term non-photochemical quenching covers a suite of processes that dissipate light energy absorbed by light harvesting antennae as heat. and is measurable as a decrease in non-photochemical quenching of at least 2%, for leaf measurements made after 40 minutes of acclimation to a light intensity of 700 μmol PAR m-2 s-1;
[0246] (h) a decrease in the ratio of the carbon isotope 12C to 13C found in either, all the dried above-ground biomass, or specific components of the above-ground biomass, leaves, reproductive structures, of at least 0.5%0 (0.5 per mille), measured as a decrease in the ratio of 12C to 13C relative to the controls with both ratio being expressed relative to the same standard; and/or
[0247] (i) an increase in the total dry weight of above-ground plant material of at least 5%.
[0248] Closely-related homologs of MYB19 derived from various diverse plant species may be overexpressed in plants and have the same functions of conferring increased photosynthetic resource use efficiency. It is thus expected that structurally similar orthologs of the MYB19 polypeptide clade, including SEQ ID NOs: 2n, where n=1-17, can confer increased yield, and/or increased vigor, biomass, or size, relative to control plants. As at least one sequence of the instant description has increased photosynthetic resource use efficiency in Arabidopsis, it is expected that the sequences provided in the Sequence Listing, or polypeptide sequences comprising one of or any of the conserved first Myb DNA binding domains provided in Table 2, or the conserved second Myb DNA binding domains provided in Table 3, will increase the photosynthetic resource use efficiency and/or yield of transgenic plants including transgenic non-Arabidopsis (plant species other than Arabidopsis species) crop or other commercially important plant species, including, but not limited to, non-Arabidopsis plants and plant species such as monocots and dicots; wheat, Setaria, corn (maize), teosinte (Zea species which is related to maize), rice, barley; rye; millet; sorghum; sugarcane, miscane, turfgrass, Miscanthus, switchgrass; soybean, cotton, rape, oilseed rape including canola, tobacco, tomato, tomatillo, potato, sunflower, alfalfa, clover, banana, blackberry, blueberry, strawberry, raspberry, cantaloupe, carrot, cauliflower, coffee, cucumber, eggplant, grapes, honeydew, lettuce, mango, melon, onion, papaya, peas, peppers, pineapple, pumpkin, spinach, squash, sweet corn, watermelon, rosaceous fruits including apple, peach, pear, cherry and plum; and brassicas including broccoli, cabbage, cauliflower, Brussels sprouts, and kohlrabi; currant; avocado; citrus fruits including oranges, lemons, grapefruit and tangerines, artichoke, cherries; endive; leek; roots such as arrowroot, beet, cassaya, turnip, radish, yam, and sweet potato; beans; woody species including pine, poplar, Eucalyptus, mint or other labiates; nuts such as walnut and peanut. Within each of these species the Closely-related homologs of MYB19 may be overexpressed or ectopically expressed in different varieties, cultivars, or germplasm.
[0249] All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.
[0250] The present invention is not limited by the specific embodiments described herein. The invention now being fully described, it will be apparent to one of ordinary skill in the art that many changes and modifications can be made thereto without departing from the spirit or scope of the appended claims. Modifications that become apparent from the foregoing description and accompanying figures fall within the scope of the claims.
Sequence CWU
1
1
1601804DNAArabidopsis thalianaAT5G52260.1 1atgaccaaat ctggagagag
accaaaacag agacagagga aagggttatg gtcacctgaa 60gaagaccaga agctcaagag
tttcatcctc tctcgtggcc atgcttgctg gaccactgtt 120cccatcctag ctggattgca
aaggaatggg aaaagctgca gattaaggtg gattaattac 180ctaagaccag gactaaagag
ggggtcgttt agtgaagaag aagaagagac catcttgact 240ttacattctt ccttgggtaa
caagtggtct cggattgcaa aatatttacc gggaagaaca 300gacaacgaga ttaagaacta
ttggcattcc tatctgaaga agagatggct caaatctcaa 360ccacaactca aaagccaaat
atcagacctc acagaatctc cttcttcact actttcttgc 420gggaaaagaa atctggaaac
cgaaacccta gatcacgtga tctccttcca gaaattttca 480gagaatccaa cttcatcacc
atccaaagaa agcaacaaca acatgatcat gaacaacagt 540aataacttgc ctaaactgtt
cttctctgag tggatcagtt cttcaaatcc acacatcgat 600tactcctctg cttttacaga
ttccaagcac attaatgaaa ctcaagatca aatcaatgaa 660gaggaagtga tgatgatcaa
taacaacaac tactcttcac ttgaggatgt catgctccgt 720acagattttt tgcagcctga
tcatgaatat gcaaattatt attcttctgg agatttcttc 780atcaacagtg accaaaatta
tgtc 8042268PRTArabidopsis
thalianaAT5G52260.1, domain AAs 17-77, 70-112 2Met Thr Lys Ser Gly Glu
Arg Pro Lys Gln Arg Gln Arg Lys Gly Leu 1 5
10 15 Trp Ser Pro Glu Glu Asp Gln Lys Leu Lys Ser
Phe Ile Leu Ser Arg 20 25
30 Gly His Ala Cys Trp Thr Thr Val Pro Ile Leu Ala Gly Leu Gln
Arg 35 40 45 Asn
Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro Gly 50
55 60 Leu Lys Arg Gly Ser Phe
Ser Glu Glu Glu Glu Glu Thr Ile Leu Thr 65 70
75 80 Leu His Ser Ser Leu Gly Asn Lys Trp Ser Arg
Ile Ala Lys Tyr Leu 85 90
95 Pro Gly Arg Thr Asp Asn Glu Ile Lys Asn Tyr Trp His Ser Tyr Leu
100 105 110 Lys Lys
Arg Trp Leu Lys Ser Gln Pro Gln Leu Lys Ser Gln Ile Ser 115
120 125 Asp Leu Thr Glu Ser Pro Ser
Ser Leu Leu Ser Cys Gly Lys Arg Asn 130 135
140 Leu Glu Thr Glu Thr Leu Asp His Val Ile Ser Phe
Gln Lys Phe Ser 145 150 155
160 Glu Asn Pro Thr Ser Ser Pro Ser Lys Glu Ser Asn Asn Asn Met Ile
165 170 175 Met Asn Asn
Ser Asn Asn Leu Pro Lys Leu Phe Phe Ser Glu Trp Ile 180
185 190 Ser Ser Ser Asn Pro His Ile Asp
Tyr Ser Ser Ala Phe Thr Asp Ser 195 200
205 Lys His Ile Asn Glu Thr Gln Asp Gln Ile Asn Glu Glu
Glu Val Met 210 215 220
Met Ile Asn Asn Asn Asn Tyr Ser Ser Leu Glu Asp Val Met Leu Arg 225
230 235 240 Thr Asp Phe Leu
Gln Pro Asp His Glu Tyr Ala Asn Tyr Tyr Ser Ser 245
250 255 Gly Asp Phe Phe Ile Asn Ser Asp Gln
Asn Tyr Val 260 265
3849DNAArabidopsis thalianaAT4G25560.1 3atggcgaaga cgaaatatgg agagagacat
aggaaagggt tatggtcacc tgaagaagac 60gagaagctaa ggagcttcat cctctcttat
ggccattctt gctggaccac tgttcccatc 120aaagctgggt tacaaaggaa tgggaagagc
tgcagattaa gatggattaa ttacctaaga 180ccagggttaa agagggatat gattagtgca
gaagaagaag agactatctt gacgtttcat 240tcttccttgg gtaacaagtg gtcgcaaata
gctaaattct taccgggaag aacagacaat 300gagataaaga actattggca ctctcatttg
aaaaagaaat ggctcaagtc tcagagctta 360caagatgcaa aatctatttc ccctccttcg
tcttcatcat catcacttgt tgcttgtgga 420aaaagaaatc cggaaacctt gatctcgaat
cacgtgttct ccttccagag acttctagag 480aacaaatctt catctccctc acaagaaagc
aacggaaata acagccatca atgttcttct 540gctcctgaga ttccaaggct tttcttctct
gaatggcttt cttcttcata tccccacacc 600gattattcct ctgagtttac cgactctaag
cacagtcaag ctccaaatgt cgaagagact 660ctctcagctt atgaagaaat gggtgatgtt
gatcagttcc attacaacga aatgatgatc 720aacaacagca actggactct taacgacatt
gtgtttggtt ccaaatgtaa gaagcaggag 780catcatattt atagagaggc ttcagattgt
aattcttctg ctgaattctt ttctccatca 840acaacgacg
8494283PRTArabidopsis
thalianaAT4G25560.1, domain AAs 15-75, 68-110 4Met Ala Lys Thr Lys Tyr
Gly Glu Arg His Arg Lys Gly Leu Trp Ser 1 5
10 15 Pro Glu Glu Asp Glu Lys Leu Arg Ser Phe Ile
Leu Ser Tyr Gly His 20 25
30 Ser Cys Trp Thr Thr Val Pro Ile Lys Ala Gly Leu Gln Arg Asn
Gly 35 40 45 Lys
Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro Gly Leu Lys 50
55 60 Arg Asp Met Ile Ser Ala
Glu Glu Glu Glu Thr Ile Leu Thr Phe His 65 70
75 80 Ser Ser Leu Gly Asn Lys Trp Ser Gln Ile Ala
Lys Phe Leu Pro Gly 85 90
95 Arg Thr Asp Asn Glu Ile Lys Asn Tyr Trp His Ser His Leu Lys Lys
100 105 110 Lys Trp
Leu Lys Ser Gln Ser Leu Gln Asp Ala Lys Ser Ile Ser Pro 115
120 125 Pro Ser Ser Ser Ser Ser Ser
Leu Val Ala Cys Gly Lys Arg Asn Pro 130 135
140 Glu Thr Leu Ile Ser Asn His Val Phe Ser Phe Gln
Arg Leu Leu Glu 145 150 155
160 Asn Lys Ser Ser Ser Pro Ser Gln Glu Ser Asn Gly Asn Asn Ser His
165 170 175 Gln Cys Ser
Ser Ala Pro Glu Ile Pro Arg Leu Phe Phe Ser Glu Trp 180
185 190 Leu Ser Ser Ser Tyr Pro His Thr
Asp Tyr Ser Ser Glu Phe Thr Asp 195 200
205 Ser Lys His Ser Gln Ala Pro Asn Val Glu Glu Thr Leu
Ser Ala Tyr 210 215 220
Glu Glu Met Gly Asp Val Asp Gln Phe His Tyr Asn Glu Met Met Ile 225
230 235 240 Asn Asn Ser Asn
Trp Thr Leu Asn Asp Ile Val Phe Gly Ser Lys Cys 245
250 255 Lys Lys Gln Glu His His Ile Tyr Arg
Glu Ala Ser Asp Cys Asn Ser 260 265
270 Ser Ala Glu Phe Phe Ser Pro Ser Thr Thr Thr 275
280 5819DNAOryza sativaLOC_Os04g45020.1
5atggggtgca aggcgtgcca gaagcccaag gtgcactacc ggaagggcct gtggtcgccg
60gaggaggacc agaagctccg cgacttcatc ctccgctacg gccacggctg ctggagcgcc
120gtccccgtga aggccgggct gcagcgtaac ggcaagagtt gcaggctgag atggatcaat
180tacctgaggc cggggctgaa gcacggcatg ttttcccgag aggaagaaga aaccgtcatg
240aacctgcacg ctacaatggg caacaagtgg tcacagatag cgcggcatct gcctggccgg
300acggacaacg aggtgaagaa ctactggaac tcgtacctca agaagcgagt cgaaggcgcg
360gaggctgcgg ccagaaaatc cgccgagccg gccgacgtcg tcaccggcag cccgaaccgc
420agcgagaccg gccaagaacg cgtcgccgct gaccggccgg cgagctccga gtcttccggg
480ccggtcgagt cgtcgtcggc cgacgactcg agcagcctca ccgagcccgc ggcggggctc
540gccgccgtcc ggccgcacgc gcccgtgatc cccaaggtca tgttcgccga ctggttcgac
600atggactacg ggactagcct cgccgggacg gcgccgggcc tgagctacca gggctcgtcg
660tcggtgcagg tcgacgtccc gtgcggcggc gccgtggact ccctgcacgg gctgggcgac
720ggcggcttct gctgggactt cgacgacgcg gccgatcaca tgcagggagg aggagggctc
780tgcgacctgc tctccatgag cgagttcctc ggcatcaac
8196273PRTOryza sativaLOC_Os04g45020.1, domain AAs 18-78, 71-113 6Met Gly
Cys Lys Ala Cys Gln Lys Pro Lys Val His Tyr Arg Lys Gly 1 5
10 15 Leu Trp Ser Pro Glu Glu Asp
Gln Lys Leu Arg Asp Phe Ile Leu Arg 20 25
30 Tyr Gly His Gly Cys Trp Ser Ala Val Pro Val Lys
Ala Gly Leu Gln 35 40 45
Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro
50 55 60 Gly Leu Lys
His Gly Met Phe Ser Arg Glu Glu Glu Glu Thr Val Met 65
70 75 80 Asn Leu His Ala Thr Met Gly
Asn Lys Trp Ser Gln Ile Ala Arg His 85
90 95 Leu Pro Gly Arg Thr Asp Asn Glu Val Lys Asn
Tyr Trp Asn Ser Tyr 100 105
110 Leu Lys Lys Arg Val Glu Gly Ala Glu Ala Ala Ala Arg Lys Ser
Ala 115 120 125 Glu
Pro Ala Asp Val Val Thr Gly Ser Pro Asn Arg Ser Glu Thr Gly 130
135 140 Gln Glu Arg Val Ala Ala
Asp Arg Pro Ala Ser Ser Glu Ser Ser Gly 145 150
155 160 Pro Val Glu Ser Ser Ser Ala Asp Asp Ser Ser
Ser Leu Thr Glu Pro 165 170
175 Ala Ala Gly Leu Ala Ala Val Arg Pro His Ala Pro Val Ile Pro Lys
180 185 190 Val Met
Phe Ala Asp Trp Phe Asp Met Asp Tyr Gly Thr Ser Leu Ala 195
200 205 Gly Thr Ala Pro Gly Leu Ser
Tyr Gln Gly Ser Ser Ser Val Gln Val 210 215
220 Asp Val Pro Cys Gly Gly Ala Val Asp Ser Leu His
Gly Leu Gly Asp 225 230 235
240 Gly Gly Phe Cys Trp Asp Phe Asp Asp Ala Ala Asp His Met Gln Gly
245 250 255 Gly Gly Gly
Leu Cys Asp Leu Leu Ser Met Ser Glu Phe Leu Gly Ile 260
265 270 Asn 7855DNABrachypodium
distachyonBradi5g16672.1 7atggggtgca agtcgtgcca gaagcccaag gcgcaccatc
ggaagggcct gtggtcgccg 60gaggaggacc agaagctccg cgactacatc atccgttatg
gccatagctg ctggagcacc 120gtccccgtca aggctggact gcagcggaac ggcaagagct
gcaggctgag atggatcaac 180tacctgaggc cggggctgaa gcacggcatg ttctcccagg
aggaagaaga gaccgtcatg 240agcctccacg ccacactggg caacaaatgg tctcggatag
cgcagcatct gccaggccgg 300accgacaacg aggtgaagaa ctactggaac tcgtacctga
agaagcgcgt ggagggcgcg 360caggcggcac cagccaaatc cgccggctcg gactcgcccc
agagcccgac ggcggcgctc 420agcgagagcg gcgttaaacg gccggagaac tccggctcgt
ccgggccgcc ggaatcgtcg 480tcggccgacg actcgagctg cctcacgggg cccgccggcg
ccgccgcggc cctgatccgg 540ccgcacgcgc ccgtgctccc caaggtcatg ttcgcggact
ggctcgacat ggacatggac 600tacggcacgg gcctgatggc gccgggcctg gacgcgggct
tcggagcggg ccggtgcagc 660agcccggccc agggcgccgc gagccagcag gggtccgtgc
aggtcgacgg cccatcgtgc 720agcgccgtgg attccttgca cgggctcggc ggcggcatct
gctgggactt cgacgcggcg 780gatcagatgc acatgcagag cgggggagga gggttctgcg
acctgctctc catgagcgag 840ttccttggga tcaac
8558285PRTBrachypodium distachyonBradi5g16672.1,
domain AAs 18-78, 71-113 8Met Gly Cys Lys Ser Cys Gln Lys Pro Lys Ala His
His Arg Lys Gly 1 5 10
15 Leu Trp Ser Pro Glu Glu Asp Gln Lys Leu Arg Asp Tyr Ile Ile Arg
20 25 30 Tyr Gly His
Ser Cys Trp Ser Thr Val Pro Val Lys Ala Gly Leu Gln 35
40 45 Arg Asn Gly Lys Ser Cys Arg Leu
Arg Trp Ile Asn Tyr Leu Arg Pro 50 55
60 Gly Leu Lys His Gly Met Phe Ser Gln Glu Glu Glu Glu
Thr Val Met 65 70 75
80 Ser Leu His Ala Thr Leu Gly Asn Lys Trp Ser Arg Ile Ala Gln His
85 90 95 Leu Pro Gly Arg
Thr Asp Asn Glu Val Lys Asn Tyr Trp Asn Ser Tyr 100
105 110 Leu Lys Lys Arg Val Glu Gly Ala Gln
Ala Ala Pro Ala Lys Ser Ala 115 120
125 Gly Ser Asp Ser Pro Gln Ser Pro Thr Ala Ala Leu Ser Glu
Ser Gly 130 135 140
Val Lys Arg Pro Glu Asn Ser Gly Ser Ser Gly Pro Pro Glu Ser Ser 145
150 155 160 Ser Ala Asp Asp Ser
Ser Cys Leu Thr Gly Pro Ala Gly Ala Ala Ala 165
170 175 Ala Leu Ile Arg Pro His Ala Pro Val Leu
Pro Lys Val Met Phe Ala 180 185
190 Asp Trp Leu Asp Met Asp Met Asp Tyr Gly Thr Gly Leu Met Ala
Pro 195 200 205 Gly
Leu Asp Ala Gly Phe Gly Ala Gly Arg Cys Ser Ser Pro Ala Gln 210
215 220 Gly Ala Ala Ser Gln Gln
Gly Ser Val Gln Val Asp Gly Pro Ser Cys 225 230
235 240 Ser Ala Val Asp Ser Leu His Gly Leu Gly Gly
Gly Ile Cys Trp Asp 245 250
255 Phe Asp Ala Ala Asp Gln Met His Met Gln Ser Gly Gly Gly Gly Phe
260 265 270 Cys Asp
Leu Leu Ser Met Ser Glu Phe Leu Gly Ile Asn 275
280 285 9873DNAZea maysGRMZM2G170049_T01 9atggggtgca
aggcgtgcga caagcccaag cccaactacc gcaagggcct gtggtcgccg 60gaggaggacc
agaagctccg cgactacatt ctcctccacg gccacggctg ctggagcgcg 120ctccccgcga
aagccgggct ccagcggaac ggcaagagct gcaggctgcg gtggatcaac 180taccttcggc
cggggctgaa gcacggcatg ttctccccgg aggaggagga gacggtgatg 240agcctccacg
ccacgctcgg caacaagtgg tccaggatcg cacggcactt gcctggcagg 300accgacaacg
aggtcaagaa ctactggaac tcgtacctca agaagagggt cgagggcaag 360gaccaggggc
ccagcacgcc cgcgccggcg gcgtccaatt cggacgacga ctcgcactgc 420gtcaagcagc
gcagggacga cgacggcacg gcggactccg gcgcgtcgga gccgcgcgag 480tcgtcgtcgg
ccgacgactc gagctgcctg acggacccgc acgcctgcag gccccacgcg 540cccgtgccgc
ccaaggtcat gttcgcggac tggctggaca tggactacgt gggcggtgcc 600ctgccggcga
cagcaccagc agcacctggt ctgctcggcg ctgcgggcgt ggccacggcc 660agcacgggcg
accgcgatca gcatcaggtg atgagcatga gccaggggtc cgttcaggtg 720gatgggccat
ccggtgccga tgtgtccctg cacggcttcg atgacagcgg cgccggctgc 780tgggagttcc
aggagcactt cgatgccatc gatcacatgc aggcggccgg cttctgcgac 840ctgctctcca
tgagcgacta cttcggcctc gac 87310291PRTZea
maysGRMZM2G170049_T01, domain AAs 18-78, 71-113 10Met Gly Cys Lys Ala Cys
Asp Lys Pro Lys Pro Asn Tyr Arg Lys Gly 1 5
10 15 Leu Trp Ser Pro Glu Glu Asp Gln Lys Leu Arg
Asp Tyr Ile Leu Leu 20 25
30 His Gly His Gly Cys Trp Ser Ala Leu Pro Ala Lys Ala Gly Leu
Gln 35 40 45 Arg
Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro 50
55 60 Gly Leu Lys His Gly Met
Phe Ser Pro Glu Glu Glu Glu Thr Val Met 65 70
75 80 Ser Leu His Ala Thr Leu Gly Asn Lys Trp Ser
Arg Ile Ala Arg His 85 90
95 Leu Pro Gly Arg Thr Asp Asn Glu Val Lys Asn Tyr Trp Asn Ser Tyr
100 105 110 Leu Lys
Lys Arg Val Glu Gly Lys Asp Gln Gly Pro Ser Thr Pro Ala 115
120 125 Pro Ala Ala Ser Asn Ser Asp
Asp Asp Ser His Cys Val Lys Gln Arg 130 135
140 Arg Asp Asp Asp Gly Thr Ala Asp Ser Gly Ala Ser
Glu Pro Arg Glu 145 150 155
160 Ser Ser Ser Ala Asp Asp Ser Ser Cys Leu Thr Asp Pro His Ala Cys
165 170 175 Arg Pro His
Ala Pro Val Pro Pro Lys Val Met Phe Ala Asp Trp Leu 180
185 190 Asp Met Asp Tyr Val Gly Gly Ala
Leu Pro Ala Thr Ala Pro Ala Ala 195 200
205 Pro Gly Leu Leu Gly Ala Ala Gly Val Ala Thr Ala Ser
Thr Gly Asp 210 215 220
Arg Asp Gln His Gln Val Met Ser Met Ser Gln Gly Ser Val Gln Val 225
230 235 240 Asp Gly Pro Ser
Gly Ala Asp Val Ser Leu His Gly Phe Asp Asp Ser 245
250 255 Gly Ala Gly Cys Trp Glu Phe Gln Glu
His Phe Asp Ala Ile Asp His 260 265
270 Met Gln Ala Ala Gly Phe Cys Asp Leu Leu Ser Met Ser Asp
Tyr Phe 275 280 285
Gly Leu Asp 290 11867DNASetaria italicaSi012304m 11atggggtgca
aggcgtgcca gaagcccaag gtgcagtacc gcaagggcct gtggtcgccg 60gaggaggacg
agaagctccg cgacttcatc ctccgctacg gccacggctg ctggagcgcg 120ctccccgcca
aggccgggct gcagcgcaac ggcaagagct gcaggctgag gtggatcaac 180tacctgaggc
cggggctgaa gcacggcatg ttctcccggg aggaggagga gaccgtcatg 240agcctccacg
ccaagcttgg caacaagtgg tctcagatcg cgcggcacct gccgggccgg 300accgacaacg
aggtgaagaa ctactggaac tcgtacctca agaagcgcgt cgagggcggc 360gcgcaggcca
agtgcgcggc ggacccggcg acacccgccg gttccgacgt ccgcgccggg 420agccccaacc
ccagcgacaa cggtcgggaa cgcgccaacc accccgcgag ctctgactcg 480tcggagccgg
tcgagtcgtc ctcggccgac gactcgagct gcctcaccgt caccgagccc 540gccagggcgg
gcgcggtgcg gccgcacgct cccgtgctcc ccaaggtcat gttcgcggac 600tggctcgaca
tggactacgg caccagcctg gcggcgctgg gtccggacgc cggcgtcttc 660gacgtgagcg
ggcgcagccc ggggcagggc ctgagccacc aggggtccgt gcaggtggac 720ggcccgtgcg
gcgcggtgga ttccctgcac gggctcggcg acggcggcat ctgcggctgg 780gggttcgacg
cggcggtgga tcagatggac gtgcagggag gagggttctg cgatctgctc 840tccatgaccg
agttccttgg gatcaac
86712289PRTSetaria italicaSi012304m, domain AAs 18-78, 71-113 12Met Gly
Cys Lys Ala Cys Gln Lys Pro Lys Val Gln Tyr Arg Lys Gly 1 5
10 15 Leu Trp Ser Pro Glu Glu Asp
Glu Lys Leu Arg Asp Phe Ile Leu Arg 20 25
30 Tyr Gly His Gly Cys Trp Ser Ala Leu Pro Ala Lys
Ala Gly Leu Gln 35 40 45
Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro
50 55 60 Gly Leu Lys
His Gly Met Phe Ser Arg Glu Glu Glu Glu Thr Val Met 65
70 75 80 Ser Leu His Ala Lys Leu Gly
Asn Lys Trp Ser Gln Ile Ala Arg His 85
90 95 Leu Pro Gly Arg Thr Asp Asn Glu Val Lys Asn
Tyr Trp Asn Ser Tyr 100 105
110 Leu Lys Lys Arg Val Glu Gly Gly Ala Gln Ala Lys Cys Ala Ala
Asp 115 120 125 Pro
Ala Thr Pro Ala Gly Ser Asp Val Arg Ala Gly Ser Pro Asn Pro 130
135 140 Ser Asp Asn Gly Arg Glu
Arg Ala Asn His Pro Ala Ser Ser Asp Ser 145 150
155 160 Ser Glu Pro Val Glu Ser Ser Ser Ala Asp Asp
Ser Ser Cys Leu Thr 165 170
175 Val Thr Glu Pro Ala Arg Ala Gly Ala Val Arg Pro His Ala Pro Val
180 185 190 Leu Pro
Lys Val Met Phe Ala Asp Trp Leu Asp Met Asp Tyr Gly Thr 195
200 205 Ser Leu Ala Ala Leu Gly Pro
Asp Ala Gly Val Phe Asp Val Ser Gly 210 215
220 Arg Ser Pro Gly Gln Gly Leu Ser His Gln Gly Ser
Val Gln Val Asp 225 230 235
240 Gly Pro Cys Gly Ala Val Asp Ser Leu His Gly Leu Gly Asp Gly Gly
245 250 255 Ile Cys Gly
Trp Gly Phe Asp Ala Ala Val Asp Gln Met Asp Val Gln 260
265 270 Gly Gly Gly Phe Cys Asp Leu Leu
Ser Met Thr Glu Phe Leu Gly Ile 275 280
285 Asn 13939DNACitrus clementinaclementine0.9_033485m
13atgggatgca agtcatcgga aaagccaatt gcaaagccga agccaaagca cagaaagggc
60ttgtggtctc ccgaagaaga ccagaggctc aagaactatg tcctccagca tggccaccct
120tgctggagct ccgtccccat caatgccggc ttgcaaagga atggaaagag ctgcagactg
180agatggatta attatttgag gccaggactt aagagagggg tgttcaatat gcaagaagaa
240gagacaatcc tgaccgtcca tcgcctgtta ggaaacaagt ggtctcaaat tgctcagcat
300ttgcctggaa gaacagataa cgagataaag aactattggc actcccattt gaagaaaaaa
360ttagccaaac ttgaagaaat ggaagcagct aatgcgacaa ctccaagctc agaaaatatg
420gaatcttcaa cttcccctaa taacaatccc tcaactcgca gctcaagcta tgaatcgttg
480caccacatgg aaaaatcatc agccggtagt actgatcagt gtgcaactca gggtcagaaa
540agttgcttgc cgaagctttt atttgcagag tggctgtcgc ttgatcatgc taatgatggt
600agcttcgcaa attccttcga gcaagtggct tccaaggaag gctttaataa taataataat
660aataataata ataataataa taataataat aataataatc agaactccaa cttggtccaa
720gattcgagtg atacatttat gaatggttac ttgtccaatg agggagcatt tggcggcgat
780tttattcata acggattcaa caacagtttt gttgatgaaa tgttgagttc aagattcaaa
840ttcgaggatc atcagttttc cggaattggg tttgttgatt ctatctctgg ggatgatgta
900tgtagtgctt tgaatatgaa taatgatgta atgtacata
93914313PRTCitrus clementinaclementine0.9_033485m, domain AAs 22-82,
75-117 14Met Gly Cys Lys Ser Ser Glu Lys Pro Ile Ala Lys Pro Lys Pro Lys
1 5 10 15 His Arg
Lys Gly Leu Trp Ser Pro Glu Glu Asp Gln Arg Leu Lys Asn 20
25 30 Tyr Val Leu Gln His Gly His
Pro Cys Trp Ser Ser Val Pro Ile Asn 35 40
45 Ala Gly Leu Gln Arg Asn Gly Lys Ser Cys Arg Leu
Arg Trp Ile Asn 50 55 60
Tyr Leu Arg Pro Gly Leu Lys Arg Gly Val Phe Asn Met Gln Glu Glu 65
70 75 80 Glu Thr Ile
Leu Thr Val His Arg Leu Leu Gly Asn Lys Trp Ser Gln 85
90 95 Ile Ala Gln His Leu Pro Gly Arg
Thr Asp Asn Glu Ile Lys Asn Tyr 100 105
110 Trp His Ser His Leu Lys Lys Lys Leu Ala Lys Leu Glu
Glu Met Glu 115 120 125
Ala Ala Asn Ala Thr Thr Pro Ser Ser Glu Asn Met Glu Ser Ser Thr 130
135 140 Ser Pro Asn Asn
Asn Pro Ser Thr Arg Ser Ser Ser Tyr Glu Ser Leu 145 150
155 160 His His Met Glu Lys Ser Ser Ala Gly
Ser Thr Asp Gln Cys Ala Thr 165 170
175 Gln Gly Gln Lys Ser Cys Leu Pro Lys Leu Leu Phe Ala Glu
Trp Leu 180 185 190
Ser Leu Asp His Ala Asn Asp Gly Ser Phe Ala Asn Ser Phe Glu Gln
195 200 205 Val Ala Ser Lys
Glu Gly Phe Asn Asn Asn Asn Asn Asn Asn Asn Asn 210
215 220 Asn Asn Asn Asn Asn Asn Asn Asn
Asn Gln Asn Ser Asn Leu Val Gln 225 230
235 240 Asp Ser Ser Asp Thr Phe Met Asn Gly Tyr Leu Ser
Asn Glu Gly Ala 245 250
255 Phe Gly Gly Asp Phe Ile His Asn Gly Phe Asn Asn Ser Phe Val Asp
260 265 270 Glu Met Leu
Ser Ser Arg Phe Lys Phe Glu Asp His Gln Phe Ser Gly 275
280 285 Ile Gly Phe Val Asp Ser Ile Ser
Gly Asp Asp Val Cys Ser Ala Leu 290 295
300 Asn Met Asn Asn Asp Val Met Tyr Ile 305
310 15885DNAPopulus trichocarpaPOPTR_0015s13190.1
15atggggtgca agtcatctga catgccaaag ctaaagccaa agccaaagca caggaaaggc
60ttgtggtcac ctgaagaaga tcaaaggctc agaaactatg tccttaaaca tggccatgga
120tgttggagct ctgtccccat taatgctggc ttgcagagga atgggaagag ctgcagacta
180agatggatta attacttgag accaggatta aaaagaggga cgttttctgc acaagaagag
240gagacaatcc tggcccttca tcacatgtta ggcaacaagt ggtctcagat agcacagcat
300ttgcctggaa gaacagataa tgagataaag aatcattggc attcctattt gaagaaaaat
360ttgctcaaag acgaagggat ggagtctttg aaaagaacaa aatctgacag ctcaaactca
420gacattatgg aactttcacc atctcccaag agactcaaaa tgcaagcttc aagttttgag
480tcatcaatga gtgcagaaaa atcatcagct gatatcaacc ggtcagttcc gcagatgttt
540gagtctccta acgaacctaa aggaagctcc ttattaccaa aggttatgtt tgcagagtgg
600ctttcacttg aaagcttcgc gagtttaggt gagcctatgg attcaaagac tacacttgat
660cataatacaa tcttccaaga caatttcttg catgattact tactggatga aagagcattt
720ggcggcgagt atcataattc actaagcgat ggttcgagcg gcgacatttt tagttcagaa
780ttcaggtttg agagccagag tccagggaat gagtttgatt ttagctctgg agaggattta
840tgtagtgact tcaacttgag caacattagt gatgtgatgt acata
88516295PRTPopulus trichocarpaPOPTR_0015s13190.1, domain AAs 22-82,
75-117 16Met Gly Cys Lys Ser Ser Asp Met Pro Lys Leu Lys Pro Lys Pro Lys
1 5 10 15 His Arg
Lys Gly Leu Trp Ser Pro Glu Glu Asp Gln Arg Leu Arg Asn 20
25 30 Tyr Val Leu Lys His Gly His
Gly Cys Trp Ser Ser Val Pro Ile Asn 35 40
45 Ala Gly Leu Gln Arg Asn Gly Lys Ser Cys Arg Leu
Arg Trp Ile Asn 50 55 60
Tyr Leu Arg Pro Gly Leu Lys Arg Gly Thr Phe Ser Ala Gln Glu Glu 65
70 75 80 Glu Thr Ile
Leu Ala Leu His His Met Leu Gly Asn Lys Trp Ser Gln 85
90 95 Ile Ala Gln His Leu Pro Gly Arg
Thr Asp Asn Glu Ile Lys Asn His 100 105
110 Trp His Ser Tyr Leu Lys Lys Asn Leu Leu Lys Asp Glu
Gly Met Glu 115 120 125
Ser Leu Lys Arg Thr Lys Ser Asp Ser Ser Asn Ser Asp Ile Met Glu 130
135 140 Leu Ser Pro Ser
Pro Lys Arg Leu Lys Met Gln Ala Ser Ser Phe Glu 145 150
155 160 Ser Ser Met Ser Ala Glu Lys Ser Ser
Ala Asp Ile Asn Arg Ser Val 165 170
175 Pro Gln Met Phe Glu Ser Pro Asn Glu Pro Lys Gly Ser Ser
Leu Leu 180 185 190
Pro Lys Val Met Phe Ala Glu Trp Leu Ser Leu Glu Ser Phe Ala Ser
195 200 205 Leu Gly Glu Pro
Met Asp Ser Lys Thr Thr Leu Asp His Asn Thr Ile 210
215 220 Phe Gln Asp Asn Phe Leu His Asp
Tyr Leu Leu Asp Glu Arg Ala Phe 225 230
235 240 Gly Gly Glu Tyr His Asn Ser Leu Ser Asp Gly Ser
Ser Gly Asp Ile 245 250
255 Phe Ser Ser Glu Phe Arg Phe Glu Ser Gln Ser Pro Gly Asn Glu Phe
260 265 270 Asp Phe Ser
Ser Gly Glu Asp Leu Cys Ser Asp Phe Asn Leu Ser Asn 275
280 285 Ile Ser Asp Val Met Tyr Ile
290 295 17807DNAEucalyptus grandisEUCGR.K00250.1
17atggcactga agtcatcaga aagaccaaaa cccaagcaca gaaagggatt gtggtcacct
60gaagaagatc agaagctcag gaactatgtc ctcaagcatg gccatggttg ctggagctct
120gtccccatta acaccggctt gcagaggaat ggcaagagct gcagattaag gtggatcaat
180tacttgaggc ctggcctaaa gagaggcatg ttcaccatgg aagaggagga gattattttt
240tcccttcatc acttgatagg caacaagtgg tctcaaatag caaagcattt gccaggaagg
300acagataacg agataaagaa tcactggcat tcttatctta agaagaaggt ggcaaacaag
360actgaatctt tatcgtcatc attagaagct caccatcatg cccggagtca atgtaccaat
420tcggacaatg tggaatcttc gcctcctcca gatcaaatcc ctccaaacca gaacccatca
480gttcatgcac catcacagga gcaaaaggaa aagacatcat ttgactttca gagggacggg
540ctacgcagct acttgcccca gattttcttc gccgagtggc tgaatcaagc tgatcaaggg
600aacaacatcc ccaattacgg cgacgctttc gatgactgct tgaatcttca ggaccccctt
660gtgcctgatc tatgcacgag tgattttgga aattcttatg gtggtgaata tgttggtagt
720gagctaagta acgggtctgc tagtgctagc gtgagcgaca tgtacagttc gcagttgaag
780ttggagatgg gatcaggttt cggggag
80718269PRTEucalyptus grandisEUCGR.K00250.1, domain AAs 18-78, 71-113
18Met Ala Leu Lys Ser Ser Glu Arg Pro Lys Pro Lys His Arg Lys Gly 1
5 10 15 Leu Trp Ser Pro
Glu Glu Asp Gln Lys Leu Arg Asn Tyr Val Leu Lys 20
25 30 His Gly His Gly Cys Trp Ser Ser Val
Pro Ile Asn Thr Gly Leu Gln 35 40
45 Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu
Arg Pro 50 55 60
Gly Leu Lys Arg Gly Met Phe Thr Met Glu Glu Glu Glu Ile Ile Phe 65
70 75 80 Ser Leu His His Leu
Ile Gly Asn Lys Trp Ser Gln Ile Ala Lys His 85
90 95 Leu Pro Gly Arg Thr Asp Asn Glu Ile Lys
Asn His Trp His Ser Tyr 100 105
110 Leu Lys Lys Lys Val Ala Asn Lys Thr Glu Ser Leu Ser Ser Ser
Leu 115 120 125 Glu
Ala His His His Ala Arg Ser Gln Cys Thr Asn Ser Asp Asn Val 130
135 140 Glu Ser Ser Pro Pro Pro
Asp Gln Ile Pro Pro Asn Gln Asn Pro Ser 145 150
155 160 Val His Ala Pro Ser Gln Glu Gln Lys Glu Lys
Thr Ser Phe Asp Phe 165 170
175 Gln Arg Asp Gly Leu Arg Ser Tyr Leu Pro Gln Ile Phe Phe Ala Glu
180 185 190 Trp Leu
Asn Gln Ala Asp Gln Gly Asn Asn Ile Pro Asn Tyr Gly Asp 195
200 205 Ala Phe Asp Asp Cys Leu Asn
Leu Gln Asp Pro Leu Val Pro Asp Leu 210 215
220 Cys Thr Ser Asp Phe Gly Asn Ser Tyr Gly Gly Glu
Tyr Val Gly Ser 225 230 235
240 Glu Leu Ser Asn Gly Ser Ala Ser Ala Ser Val Ser Asp Met Tyr Ser
245 250 255 Ser Gln Leu
Lys Leu Glu Met Gly Ser Gly Phe Gly Glu 260
265 19885DNAEucalyptus grandisEUCGR.K00251.1 19atggcattga
agtcatcaga aaggccaaag cccaagcaca ggaagggctt gtggtcacct 60gaagaagacc
agaggctcag gaactatatc ctgaaccatg gccatggtta ctggagctct 120gtccccatta
acaccggctt gcagaggaat ggcaagagct gcagattaag gtggatcaat 180tacttgaggc
ctggcctaaa gagaggcatg ttcaccctgg aagaagagga gattattttg 240tcccttcatc
gcttgatagg caataagtgg tctcaaatag caaagcattt gccaggaagg 300accgataatg
agataaagaa tcactggcat tcttatctta agaagaaggt ggcaaataag 360actgaatcat
cgtcatcatc agaagcccgc cataatgccc agagtcaatg taccaattcg 420gacaatgtgg
aatcttcgcc ttcaccagat caaatcccca ctaaccaaaa cgcatcagtt 480catgcaccgt
cacaggaaca aaaggaaaag atgtcattgg actttccgaa tgggggtcca 540cgcagctgct
tgcccaatat tttcttcgcc gagtggctga atcaagctga tcaagggtac 600aacgttccga
cctatggcga tgctttcgat tatcgctcaa atattcagga ctctcttgtg 660catgattggt
gcacaagcga ttttgggaat tctaatggcg gtgagtatgt tgggaatgag 720ctaagtaagg
ggtccgctag tgccagcgtg agcgacatgt acagttcgcg gttgaagtcg 780gagatggatc
aggtttcggg aggtgggttt tacttggatt atttctctgg ggatgatatc 840tgtagtcagt
tcgacatggg cagtgatgta aatatgatgt acata
88520295PRTEucalyptus grandisEUCGR.K00251.1, domain AAs 18-78, 71-113
20Met Ala Leu Lys Ser Ser Glu Arg Pro Lys Pro Lys His Arg Lys Gly 1
5 10 15 Leu Trp Ser Pro
Glu Glu Asp Gln Arg Leu Arg Asn Tyr Ile Leu Asn 20
25 30 His Gly His Gly Tyr Trp Ser Ser Val
Pro Ile Asn Thr Gly Leu Gln 35 40
45 Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu
Arg Pro 50 55 60
Gly Leu Lys Arg Gly Met Phe Thr Leu Glu Glu Glu Glu Ile Ile Leu 65
70 75 80 Ser Leu His Arg Leu
Ile Gly Asn Lys Trp Ser Gln Ile Ala Lys His 85
90 95 Leu Pro Gly Arg Thr Asp Asn Glu Ile Lys
Asn His Trp His Ser Tyr 100 105
110 Leu Lys Lys Lys Val Ala Asn Lys Thr Glu Ser Ser Ser Ser Ser
Glu 115 120 125 Ala
Arg His Asn Ala Gln Ser Gln Cys Thr Asn Ser Asp Asn Val Glu 130
135 140 Ser Ser Pro Ser Pro Asp
Gln Ile Pro Thr Asn Gln Asn Ala Ser Val 145 150
155 160 His Ala Pro Ser Gln Glu Gln Lys Glu Lys Met
Ser Leu Asp Phe Pro 165 170
175 Asn Gly Gly Pro Arg Ser Cys Leu Pro Asn Ile Phe Phe Ala Glu Trp
180 185 190 Leu Asn
Gln Ala Asp Gln Gly Tyr Asn Val Pro Thr Tyr Gly Asp Ala 195
200 205 Phe Asp Tyr Arg Ser Asn Ile
Gln Asp Ser Leu Val His Asp Trp Cys 210 215
220 Thr Ser Asp Phe Gly Asn Ser Asn Gly Gly Glu Tyr
Val Gly Asn Glu 225 230 235
240 Leu Ser Lys Gly Ser Ala Ser Ala Ser Val Ser Asp Met Tyr Ser Ser
245 250 255 Arg Leu Lys
Ser Glu Met Asp Gln Val Ser Gly Gly Gly Phe Tyr Leu 260
265 270 Asp Tyr Phe Ser Gly Asp Asp Ile
Cys Ser Gln Phe Asp Met Gly Ser 275 280
285 Asp Val Asn Met Met Tyr Ile 290
295 21975DNAPopulus trichocarpaPOPTR_0012s13260.1 21atgccaaagg cattcattgc
atccatcaca aagtccaaga ctctctttct cttgtacaag 60tcaccaatcc ttctcatcat
cggtgttctt ggcgaaatgg ggtgcaaatc atcagacaag 120ccaaagccaa agctaaggca
caggaaaggc ttgtggtcac ctgaagaaga tcaaaggctt 180ggaagctatg tctttcaaca
tggccacgga tgttggagct ctgtccccat taatgctggc 240ttgcagagga ctgggaagag
ctgcagatta agatggatta attacttgag accaggactg 300aaaagagggg cgttttctac
agacgaagaa gagacaatcc tgacccttca tcgcatgtta 360ggcaacaagt ggtctcaaat
tgcacagcat ttgcctggaa gaacagacaa tgagataaag 420aaccattggc attcctattt
gaagaaaaag ttgttcaaag ctgaaggaat ggaatctcct 480aataagactc aatctgccag
ctcaaactca gacaatatgg atctttcacc ctctcccaaa 540aggcttaaaa tgcaaagtcc
tgaatcgtca atgaatatgg aaaaaccatc aactgatatc 600gaccggccgg tacttccacg
gatgtttgac tatcttaaag aacctaacag aagctcctta 660ttaccaaagg ttatgtttgc
tgagtggctc tcacttgaca gctttgcaag ttcaggtgag 720cctgtggttt caaagagtac
attcgatcat aatccaagct tccaagacac tagtttcatg 780catcattact tactggaaga
aggagcattt ggtggcgact atcaaaattc tctaagcgat 840ggttcgagcg gcgacatttt
tagttcagaa ttcaaatttg aaagccagag tccaggaaat 900gagtttgatt ttagctctgg
agaggattta tgtagggaat tcaacttccg taatattggt 960gatgtgatgt acata
97522325PRTPopulus
trichocarpaPOPTR_0012s13260.1, domain AAs 52-112, 105-147 22Met Pro Lys
Ala Phe Ile Ala Ser Ile Thr Lys Ser Lys Thr Leu Phe 1 5
10 15 Leu Leu Tyr Lys Ser Pro Ile Leu
Leu Ile Ile Gly Val Leu Gly Glu 20 25
30 Met Gly Cys Lys Ser Ser Asp Lys Pro Lys Pro Lys Leu
Arg His Arg 35 40 45
Lys Gly Leu Trp Ser Pro Glu Glu Asp Gln Arg Leu Gly Ser Tyr Val 50
55 60 Phe Gln His Gly
His Gly Cys Trp Ser Ser Val Pro Ile Asn Ala Gly 65 70
75 80 Leu Gln Arg Thr Gly Lys Ser Cys Arg
Leu Arg Trp Ile Asn Tyr Leu 85 90
95 Arg Pro Gly Leu Lys Arg Gly Ala Phe Ser Thr Asp Glu Glu
Glu Thr 100 105 110
Ile Leu Thr Leu His Arg Met Leu Gly Asn Lys Trp Ser Gln Ile Ala
115 120 125 Gln His Leu Pro
Gly Arg Thr Asp Asn Glu Ile Lys Asn His Trp His 130
135 140 Ser Tyr Leu Lys Lys Lys Leu Phe
Lys Ala Glu Gly Met Glu Ser Pro 145 150
155 160 Asn Lys Thr Gln Ser Ala Ser Ser Asn Ser Asp Asn
Met Asp Leu Ser 165 170
175 Pro Ser Pro Lys Arg Leu Lys Met Gln Ser Pro Glu Ser Ser Met Asn
180 185 190 Met Glu Lys
Pro Ser Thr Asp Ile Asp Arg Pro Val Leu Pro Arg Met 195
200 205 Phe Asp Tyr Leu Lys Glu Pro Asn
Arg Ser Ser Leu Leu Pro Lys Val 210 215
220 Met Phe Ala Glu Trp Leu Ser Leu Asp Ser Phe Ala Ser
Ser Gly Glu 225 230 235
240 Pro Val Val Ser Lys Ser Thr Phe Asp His Asn Pro Ser Phe Gln Asp
245 250 255 Thr Ser Phe Met
His His Tyr Leu Leu Glu Glu Gly Ala Phe Gly Gly 260
265 270 Asp Tyr Gln Asn Ser Leu Ser Asp Gly
Ser Ser Gly Asp Ile Phe Ser 275 280
285 Ser Glu Phe Lys Phe Glu Ser Gln Ser Pro Gly Asn Glu Phe
Asp Phe 290 295 300
Ser Ser Gly Glu Asp Leu Cys Arg Glu Phe Asn Phe Arg Asn Ile Gly 305
310 315 320 Asp Val Met Tyr Ile
325 23873DNAGlycine maxGlyma16g31280.1 23atggagagcc
agccactaga aaaagcaaaa ccaaaataca gaaaaggctt atggtcacct 60gaagaagata
ataaactcag aaaccatatc attaagcatg gtcatggctg ctggagctct 120gtccctatta
aggcaggctt gcaaagaaat gggaagagct gtagactaag gtggattaac 180tacttgaggc
caggattgaa gagaggggtg ttcagcaaac atgaggaaga tacaatcatg 240gtcctacacc
atatgttagg aaacaagtgg tctcaaatag cacagcattt gccaggaagg 300actgacaatg
agataaaaaa ttattggcat tcatatttga aaaagaaaga gatcaaagcc 360aaggaaatgg
aatctgataa agaaattcag catgctagct caagttcaga cacaatggaa 420aactcactct
ctcctcagaa acttgcaaca caagatccaa gttatagttt gttagaaaac 480ctggacaaat
caatagcaca caatgataac tttttctcac aaagctataa cttttccaag 540gaggcttgtc
agagttccct accattacca aaactcctat tttctgagtg gctttcagtg 600gatcaagtag
atggtggaag ctcagtgaat tctgatgatt ccttggtctt ggggaatgaa 660tttgatcaaa
attcaacttt ccaagaagct ataatgcata tgttagaaga aaactttggt 720gaagagtatc
ataatagtct aattcacagt tcaaccactg aggtctacaa ttcacaacta 780aagtcaacaa
atcaagtgga tggaagtgac ttcatcaatt gtattcccgg gaatgagttg 840tgtagcaatt
tcagcctaac caatcatgct atg
87324291PRTGlycine maxGlyma16g31280.1, domain AAs 18-78, 71-113 24Met Glu
Ser Gln Pro Leu Glu Lys Ala Lys Pro Lys Tyr Arg Lys Gly 1 5
10 15 Leu Trp Ser Pro Glu Glu Asp
Asn Lys Leu Arg Asn His Ile Ile Lys 20 25
30 His Gly His Gly Cys Trp Ser Ser Val Pro Ile Lys
Ala Gly Leu Gln 35 40 45
Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro
50 55 60 Gly Leu Lys
Arg Gly Val Phe Ser Lys His Glu Glu Asp Thr Ile Met 65
70 75 80 Val Leu His His Met Leu Gly
Asn Lys Trp Ser Gln Ile Ala Gln His 85
90 95 Leu Pro Gly Arg Thr Asp Asn Glu Ile Lys Asn
Tyr Trp His Ser Tyr 100 105
110 Leu Lys Lys Lys Glu Ile Lys Ala Lys Glu Met Glu Ser Asp Lys
Glu 115 120 125 Ile
Gln His Ala Ser Ser Ser Ser Asp Thr Met Glu Asn Ser Leu Ser 130
135 140 Pro Gln Lys Leu Ala Thr
Gln Asp Pro Ser Tyr Ser Leu Leu Glu Asn 145 150
155 160 Leu Asp Lys Ser Ile Ala His Asn Asp Asn Phe
Phe Ser Gln Ser Tyr 165 170
175 Asn Phe Ser Lys Glu Ala Cys Gln Ser Ser Leu Pro Leu Pro Lys Leu
180 185 190 Leu Phe
Ser Glu Trp Leu Ser Val Asp Gln Val Asp Gly Gly Ser Ser 195
200 205 Val Asn Ser Asp Asp Ser Leu
Val Leu Gly Asn Glu Phe Asp Gln Asn 210 215
220 Ser Thr Phe Gln Glu Ala Ile Met His Met Leu Glu
Glu Asn Phe Gly 225 230 235
240 Glu Glu Tyr His Asn Ser Leu Ile His Ser Ser Thr Thr Glu Val Tyr
245 250 255 Asn Ser Gln
Leu Lys Ser Thr Asn Gln Val Asp Gly Ser Asp Phe Ile 260
265 270 Asn Cys Ile Pro Gly Asn Glu Leu
Cys Ser Asn Phe Ser Leu Thr Asn 275 280
285 His Ala Met 290 25786DNAGlycine
maxGlyma09g25590.1 25atggagagca agccactaga aaaagcaaaa ccaaaataca
gaaagggctt atggtcacca 60gaagaagata ataagctcag aaatcatatc attaagcatg
gtcatggctg ctggagctct 120gtccctatta aggcaggctt gcaaagaaat gggaagagct
gcagactaag gtggattaac 180tacttgaggc caggattgaa gagaggggtg ttcagcaaac
atgagaaaga tacaatcatg 240gccctacacc atatgttagg aaacaagtgg tctcagatag
cacagcattt gccaggaagg 300actgacaatg aggtaaaaaa ttactggcat tcatatttga
aaaagaaagt catcaaagct 360aaggaaatgg aatctgataa acaaattcaa catgccggct
caagttcaga cacagtggaa 420aacgcactct ctcctcagaa acttgcaaca caagattcaa
gttatgggtt gttagaaaac 480cttgacaaat caatagcaca aaatgataac tttttctcga
aaagctataa cttttccaag 540gaggcttatc agagttctct accactacca aaactcttat
tttctgagtg gctatcagtg 600gatcaagagt atcataatcg tctaattcac agttcaacca
ctgaggtcta taattcacaa 660ataaagtcaa caaatcaaat ggatggaagt gatttcatga
attgtattcc cgggaatgag 720ttacgtagca atttcagcct aaccaatcat ggtgagttgg
aaggagaaga atacaatgca 780attcct
78626262PRTGlycine maxGlyma09g25590.1, domain AAs
18-78, 71-113 26Met Glu Ser Lys Pro Leu Glu Lys Ala Lys Pro Lys Tyr Arg
Lys Gly 1 5 10 15
Leu Trp Ser Pro Glu Glu Asp Asn Lys Leu Arg Asn His Ile Ile Lys
20 25 30 His Gly His Gly Cys
Trp Ser Ser Val Pro Ile Lys Ala Gly Leu Gln 35
40 45 Arg Asn Gly Lys Ser Cys Arg Leu Arg
Trp Ile Asn Tyr Leu Arg Pro 50 55
60 Gly Leu Lys Arg Gly Val Phe Ser Lys His Glu Lys Asp
Thr Ile Met 65 70 75
80 Ala Leu His His Met Leu Gly Asn Lys Trp Ser Gln Ile Ala Gln His
85 90 95 Leu Pro Gly Arg
Thr Asp Asn Glu Val Lys Asn Tyr Trp His Ser Tyr 100
105 110 Leu Lys Lys Lys Val Ile Lys Ala Lys
Glu Met Glu Ser Asp Lys Gln 115 120
125 Ile Gln His Ala Gly Ser Ser Ser Asp Thr Val Glu Asn Ala
Leu Ser 130 135 140
Pro Gln Lys Leu Ala Thr Gln Asp Ser Ser Tyr Gly Leu Leu Glu Asn 145
150 155 160 Leu Asp Lys Ser Ile
Ala Gln Asn Asp Asn Phe Phe Ser Lys Ser Tyr 165
170 175 Asn Phe Ser Lys Glu Ala Tyr Gln Ser Ser
Leu Pro Leu Pro Lys Leu 180 185
190 Leu Phe Ser Glu Trp Leu Ser Val Asp Gln Glu Tyr His Asn Arg
Leu 195 200 205 Ile
His Ser Ser Thr Thr Glu Val Tyr Asn Ser Gln Ile Lys Ser Thr 210
215 220 Asn Gln Met Asp Gly Ser
Asp Phe Met Asn Cys Ile Pro Gly Asn Glu 225 230
235 240 Leu Arg Ser Asn Phe Ser Leu Thr Asn His Gly
Glu Leu Glu Gly Glu 245 250
255 Glu Tyr Asn Ala Ile Pro 260 27837DNASolanum
lycopersicumSolyc03g025870.2.1 27atggggtgca aattggcagc tgagaagcca
aaacaaaaac acaagaaggg attatggtct 60cctgatgaag atgataggct caaaaattat
atgattaagc atggtcatgg atgttggagc 120tctgttccca ttaatgctgg cttgcaaaga
aatggaaaga gttgtagact gagatggatt 180aattatttaa ggcctggctt aaagagaggg
gcatttagct tagaagagga agacataata 240ttgacccttc atgccatgtt tggcaacaaa
tggtctcaga ttgcacaaca gttacctgga 300agaacggata acgagataaa gaatcactgg
cactcgtatt taaagaaaag agtgtccaaa 360atgggagaaa atgaagggca cactaagcct
gggaaaacag attcttcttc accttcttta 420aagaaattga ctccacagaa ttcaagtttg
gattcatttg aacatattga aggatcatta 480gcagattcag atcaatctgt ttatccaaga
gagactcaaa agagtaattt acctaaagta 540ttattcgcgg aatggctttc gttggatcag
tttaatggac aagattttca aaactcaggg 600agtttcagtt ttgaaccttg caagagtaac
tttgtgtata ataataatgc agagttacat 660gacatactca tgcatagttt accgatgaac
aacgatgatg ggaatggcgt aaatcaagag 720gttcttcaca atgatatttt cccaccacaa
ctcaagtttg aggatacatt gtctggtaac 780ggatttgagg agtttatgtc aagggagttc
aatattaacg acgatgtgat gtacata 83728279PRTSolanum
lycopersicumSolyc03g025870.2.1, domain AAs 19-79, 72-114 28Met Gly Cys
Lys Leu Ala Ala Glu Lys Pro Lys Gln Lys His Lys Lys 1 5
10 15 Gly Leu Trp Ser Pro Asp Glu Asp
Asp Arg Leu Lys Asn Tyr Met Ile 20 25
30 Lys His Gly His Gly Cys Trp Ser Ser Val Pro Ile Asn
Ala Gly Leu 35 40 45
Gln Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg 50
55 60 Pro Gly Leu Lys
Arg Gly Ala Phe Ser Leu Glu Glu Glu Asp Ile Ile 65 70
75 80 Leu Thr Leu His Ala Met Phe Gly Asn
Lys Trp Ser Gln Ile Ala Gln 85 90
95 Gln Leu Pro Gly Arg Thr Asp Asn Glu Ile Lys Asn His Trp
His Ser 100 105 110
Tyr Leu Lys Lys Arg Val Ser Lys Met Gly Glu Asn Glu Gly His Thr
115 120 125 Lys Pro Gly Lys
Thr Asp Ser Ser Ser Pro Ser Leu Lys Lys Leu Thr 130
135 140 Pro Gln Asn Ser Ser Leu Asp Ser
Phe Glu His Ile Glu Gly Ser Leu 145 150
155 160 Ala Asp Ser Asp Gln Ser Val Tyr Pro Arg Glu Thr
Gln Lys Ser Asn 165 170
175 Leu Pro Lys Val Leu Phe Ala Glu Trp Leu Ser Leu Asp Gln Phe Asn
180 185 190 Gly Gln Asp
Phe Gln Asn Ser Gly Ser Phe Ser Phe Glu Pro Cys Lys 195
200 205 Ser Asn Phe Val Tyr Asn Asn Asn
Ala Glu Leu His Asp Ile Leu Met 210 215
220 His Ser Leu Pro Met Asn Asn Asp Asp Gly Asn Gly Val
Asn Gln Glu 225 230 235
240 Val Leu His Asn Asp Ile Phe Pro Pro Gln Leu Lys Phe Glu Asp Thr
245 250 255 Leu Ser Gly Asn
Gly Phe Glu Glu Phe Met Ser Arg Glu Phe Asn Ile 260
265 270 Asn Asp Asp Val Met Tyr Ile
275 29894DNAVitis viniferaGSVIVT01028984001 29atggggtgta
attcattgga gaagtcgaag accaagccca aacaccgaaa ggggttatgg 60tcaccggaag
aagatgctag gctcagaaac tatgtcctca aatatggcct tggctgctgg 120agctccgtcc
ctgttaacgc cggtttgcaa aggaatggaa agagctgcag attaaggtgg 180attaactact
taagaccagg attaaaacgc gggatgttta cgatcgagga ggaagagacg 240atcatggccc
ttcatcgctt gttaggcaac aagtggtctc agatagcgca gaattttcct 300ggaagaactg
ataatgagat taagaactac tggcattcat gtctcaagaa gaaagtggtg 360aaagctcagg
aaatggaagt tcatatgaac tcccaatgca tcaactctaa ctcacagagc 420attgattctt
caacttcaca agaaaagcca tcaatccaac ttccgggttt cgaatcgttt 480gaaaacatga
aaggatcatc ttcaacagat actgatcagt ccattccaca gatgttggac 540tgtcctagag
tggacacgca ggaaagcccc ttgccgaaga ttttattcgc agagtggctt 600tctcttgacc
atatttacgg ccagctcttt gttaattcag gcgagtcagt catttccaag 660gatactcttg
atcagcatga tccaaccttt caagacaatt tcacgcatgg tttcctactg 720aacgaggagt
catatgtagg tgaattgcac catggcctaa gcaatgattc gtccagtgac 780atgttttcgc
cgcaatttaa gttcgagagc cagactccgg gaagtgggat atgtgatttt 840gtgtatgggg
atgaaatatg cagtgatttc aacatgaacg gccatgtaat gtac
89430298PRTVitis viniferaGSVIVT01028984001, domain AAs 20-80, 73-115
30Met Gly Cys Asn Ser Leu Glu Lys Ser Lys Thr Lys Pro Lys His Arg 1
5 10 15 Lys Gly Leu Trp
Ser Pro Glu Glu Asp Ala Arg Leu Arg Asn Tyr Val 20
25 30 Leu Lys Tyr Gly Leu Gly Cys Trp Ser
Ser Val Pro Val Asn Ala Gly 35 40
45 Leu Gln Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn
Tyr Leu 50 55 60
Arg Pro Gly Leu Lys Arg Gly Met Phe Thr Ile Glu Glu Glu Glu Thr 65
70 75 80 Ile Met Ala Leu His
Arg Leu Leu Gly Asn Lys Trp Ser Gln Ile Ala 85
90 95 Gln Asn Phe Pro Gly Arg Thr Asp Asn Glu
Ile Lys Asn Tyr Trp His 100 105
110 Ser Cys Leu Lys Lys Lys Val Val Lys Ala Gln Glu Met Glu Val
His 115 120 125 Met
Asn Ser Gln Cys Ile Asn Ser Asn Ser Gln Ser Ile Asp Ser Ser 130
135 140 Thr Ser Gln Glu Lys Pro
Ser Ile Gln Leu Pro Gly Phe Glu Ser Phe 145 150
155 160 Glu Asn Met Lys Gly Ser Ser Ser Thr Asp Thr
Asp Gln Ser Ile Pro 165 170
175 Gln Met Leu Asp Cys Pro Arg Val Asp Thr Gln Glu Ser Pro Leu Pro
180 185 190 Lys Ile
Leu Phe Ala Glu Trp Leu Ser Leu Asp His Ile Tyr Gly Gln 195
200 205 Leu Phe Val Asn Ser Gly Glu
Ser Val Ile Ser Lys Asp Thr Leu Asp 210 215
220 Gln His Asp Pro Thr Phe Gln Asp Asn Phe Thr His
Gly Phe Leu Leu 225 230 235
240 Asn Glu Glu Ser Tyr Val Gly Glu Leu His His Gly Leu Ser Asn Asp
245 250 255 Ser Ser Ser
Asp Met Phe Ser Pro Gln Phe Lys Phe Glu Ser Gln Thr 260
265 270 Pro Gly Ser Gly Ile Cys Asp Phe
Val Tyr Gly Asp Glu Ile Cys Ser 275 280
285 Asp Phe Asn Met Asn Gly His Val Met Tyr 290
295 31723DNAEucalyptus grandisEUCGR.A02796.1
31atgggatgca agtcagtgga aaaaccaaag gcaaggcaca gaaaggggtt gtggtcacca
60gatgaagacc agaggctcag aaactacatc cataaacacg gctacagttg ctggagctca
120gttcccatca atgcaggttt gcagaggaat ggtaagagct gcagattaag gtggattaat
180tacctgaggc caggattaaa gagaggcgcg ttcacagtac aggaggaaga gacaattttg
240aacctccacc acttgttagg caacaagtgg tctcaaatag cacagcatct ccctggaagg
300accgataacg aaataaagaa tcattggcat tcttatctta agaagaagat taatatcaaa
360gctgacgaat ctcaccttca gatgccgagc aattcagact ctgtgggatc tccgaactct
420acggactttc catctgatca cacccaaagt tctgatacaa cggaatatac aaaaagctca
480gcttctcaaa ttccgaaaat tttcaacccc acaaaagagg gtgaaagctc attgccgacg
540cttctttttg aggagtggct ttctctggat aattctcctg gaggaagttt cacaaaccat
600gctgaatcac aagatcaaat ttcgggaaac gggcttgtgc agtgtctttc tggggatgat
660ctttgtatcc tttccaagtc aggctgttca cctgaagaag tgattagccg agaaatattg
720aaa
72332241PRTEucalyptus grandisEUCGR.A02796.1, domain AAs 18-78, 71-113
32Met Gly Cys Lys Ser Val Glu Lys Pro Lys Ala Arg His Arg Lys Gly 1
5 10 15 Leu Trp Ser Pro
Asp Glu Asp Gln Arg Leu Arg Asn Tyr Ile His Lys 20
25 30 His Gly Tyr Ser Cys Trp Ser Ser Val
Pro Ile Asn Ala Gly Leu Gln 35 40
45 Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu
Arg Pro 50 55 60
Gly Leu Lys Arg Gly Ala Phe Thr Val Gln Glu Glu Glu Thr Ile Leu 65
70 75 80 Asn Leu His His Leu
Leu Gly Asn Lys Trp Ser Gln Ile Ala Gln His 85
90 95 Leu Pro Gly Arg Thr Asp Asn Glu Ile Lys
Asn His Trp His Ser Tyr 100 105
110 Leu Lys Lys Lys Ile Asn Ile Lys Ala Asp Glu Ser His Leu Gln
Met 115 120 125 Pro
Ser Asn Ser Asp Ser Val Gly Ser Pro Asn Ser Thr Asp Phe Pro 130
135 140 Ser Asp His Thr Gln Ser
Ser Asp Thr Thr Glu Tyr Thr Lys Ser Ser 145 150
155 160 Ala Ser Gln Ile Pro Lys Ile Phe Asn Pro Thr
Lys Glu Gly Glu Ser 165 170
175 Ser Leu Pro Thr Leu Leu Phe Glu Glu Trp Leu Ser Leu Asp Asn Ser
180 185 190 Pro Gly
Gly Ser Phe Thr Asn His Ala Glu Ser Gln Asp Gln Ile Ser 195
200 205 Gly Asn Gly Leu Val Gln Cys
Leu Ser Gly Asp Asp Leu Cys Ile Leu 210 215
220 Ser Lys Ser Gly Cys Ser Pro Glu Glu Val Ile Ser
Arg Glu Ile Leu 225 230 235
240 Lys 33783DNAArabidopsis thalianaAT3G48920.1 33atggtgttta aatcagaaaa
atcaaaccgg gaaatgaaat caaaggagaa gcaaaggaag 60ggattatggt cacccgagga
agatgagaag cttaggagtc atgtcctcaa atatggccat 120ggatgctgga gtactattcc
tcttcaagct ggattgcaga ggaatgggaa gagttgtaga 180ttaaggtggg ttaattattt
aagacctgga cttaagaagt ctttattcac taaacaagag 240gaaactatac ttctttcact
tcattccatg ttgggtaaca aatggtctca gatatcgaaa 300ttcttaccag gaagaaccga
caacgagatc aaaaactatt ggcattctaa tctaaagaag 360ggtgtaactt tgaaacaaca
tgaaaccaca aaaaaacatc aaacaccttt aatcacaaac 420tcacttgagg ccttgcagag
ttcaactgaa agatcttctt catctatcaa tgtcggagaa 480acgtctaatg ctcaaacctc
aagcttttcg ccaaatctcg tgttctcgga atggttagat 540catagtttgc ttatggatca
gtcacctcaa aagtctagct atgttcaaaa tcttgtttta 600ccggaagaga gaggattcat
tggaccatgt ggccctcgtt atttgggaaa cgactctttg 660cctgatttcg tgccaaattc
agaatttttg ttggatgatg agatatcatc tgagatcgag 720ttctgtactt cattttcaga
caactttttg ttcgatggtc tcatcaacga gctacgacca 780atg
78334261PRTArabidopsis
thalianaAT3G48920.1, domain AAs 23-83, 76-118 34Met Val Phe Lys Ser Glu
Lys Ser Asn Arg Glu Met Lys Ser Lys Glu 1 5
10 15 Lys Gln Arg Lys Gly Leu Trp Ser Pro Glu Glu
Asp Glu Lys Leu Arg 20 25
30 Ser His Val Leu Lys Tyr Gly His Gly Cys Trp Ser Thr Ile Pro
Leu 35 40 45 Gln
Ala Gly Leu Gln Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Val 50
55 60 Asn Tyr Leu Arg Pro Gly
Leu Lys Lys Ser Leu Phe Thr Lys Gln Glu 65 70
75 80 Glu Thr Ile Leu Leu Ser Leu His Ser Met Leu
Gly Asn Lys Trp Ser 85 90
95 Gln Ile Ser Lys Phe Leu Pro Gly Arg Thr Asp Asn Glu Ile Lys Asn
100 105 110 Tyr Trp
His Ser Asn Leu Lys Lys Gly Val Thr Leu Lys Gln His Glu 115
120 125 Thr Thr Lys Lys His Gln Thr
Pro Leu Ile Thr Asn Ser Leu Glu Ala 130 135
140 Leu Gln Ser Ser Thr Glu Arg Ser Ser Ser Ser Ile
Asn Val Gly Glu 145 150 155
160 Thr Ser Asn Ala Gln Thr Ser Ser Phe Ser Pro Asn Leu Val Phe Ser
165 170 175 Glu Trp Leu
Asp His Ser Leu Leu Met Asp Gln Ser Pro Gln Lys Ser 180
185 190 Ser Tyr Val Gln Asn Leu Val Leu
Pro Glu Glu Arg Gly Phe Ile Gly 195 200
205 Pro Cys Gly Pro Arg Tyr Leu Gly Asn Asp Ser Leu Pro
Asp Phe Val 210 215 220
Pro Asn Ser Glu Phe Leu Leu Asp Asp Glu Ile Ser Ser Glu Ile Glu 225
230 235 240 Phe Cys Thr Ser
Phe Ser Asp Asn Phe Leu Phe Asp Gly Leu Ile Asn 245
250 255 Glu Leu Arg Pro Met 260
351059DNASolanum lycopersicumSolyc11g065840.1.1 35atgaggaagc
ctgagttctc ctcctcctcc tcttcctcct ccgcaaagaa caataacaat 60aacaataata
ataacacgaa cgtgaagcta agaaaagggt tgtggtctcc agaggaagat 120gaaaagctta
tgcattatat gctaacaaat ggacaagggt gttggagtga tgtagcaaga 180aatgctggat
tacaaagatg tggaaagagt tgtagactca gatggatcaa ttatttgagg 240ccagatctta
agagaggtgc attttcacct caagaagaag aacatattat ccatttacat 300tccattcttg
gtaacaggtg gtctcaaata gctgcacgtt tgcctggacg tactgataat 360gaaatcaaga
atttttggaa ttcgacattg aaaaagaggc taaagaactc atcatcatct 420tctacaccat
caccaaatgc aagtgattca tcctcagatc atccctccaa agaactcaat 480atgggagtca
ctcaacaagg attcatgcca atgctcaaac ataacctaat gtccatgtac 540atggattcaa
ccacctctcc ttcttcctcg tctatagccc taaataccat aaatattgat 600cctttgccca
ccctcgaaca caccttaata aacatgccta atggattcaa cgcgccctca 660tacttgagta
ctacacaacc atgcttggta caaggtggga atattgtgag tgctaatggt 720ggaaatcttt
tttatgggaa taaccatggg atatttggag ggaatcttag tatggaaggt 780catgaactct
atgttccacc attggagaat gtaagtattg agtatcaaaa tgttgaaaat 840gggaatttta
gtcatcatca aaacaacaat aaccctaaca acatgaccaa cttgatcaat 900actagccata
atttcaatac ttgtagtaat atcaaagtag aaaattttgg agggataggg 960aattattggg
aaggagatga cctaaaagtg ggagagtggg acttggagga attgatgaag 1020gatgtttcac
ctttcccttt tcttgatttc caagttgaa
105936353PRTSolanum lycopersicumSolyc11g065840.1.1 36Met Arg Lys Pro Glu
Phe Ser Ser Ser Ser Ser Ser Ser Ser Ala Lys 1 5
10 15 Asn Asn Asn Asn Asn Asn Asn Asn Asn Thr
Asn Val Lys Leu Arg Lys 20 25
30 Gly Leu Trp Ser Pro Glu Glu Asp Glu Lys Leu Met His Tyr Met
Leu 35 40 45 Thr
Asn Gly Gln Gly Cys Trp Ser Asp Val Ala Arg Asn Ala Gly Leu 50
55 60 Gln Arg Cys Gly Lys Ser
Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg 65 70
75 80 Pro Asp Leu Lys Arg Gly Ala Phe Ser Pro Gln
Glu Glu Glu His Ile 85 90
95 Ile His Leu His Ser Ile Leu Gly Asn Arg Trp Ser Gln Ile Ala Ala
100 105 110 Arg Leu
Pro Gly Arg Thr Asp Asn Glu Ile Lys Asn Phe Trp Asn Ser 115
120 125 Thr Leu Lys Lys Arg Leu Lys
Asn Ser Ser Ser Ser Ser Thr Pro Ser 130 135
140 Pro Asn Ala Ser Asp Ser Ser Ser Asp His Pro Ser
Lys Glu Leu Asn 145 150 155
160 Met Gly Val Thr Gln Gln Gly Phe Met Pro Met Leu Lys His Asn Leu
165 170 175 Met Ser Met
Tyr Met Asp Ser Thr Thr Ser Pro Ser Ser Ser Ser Ile 180
185 190 Ala Leu Asn Thr Ile Asn Ile Asp
Pro Leu Pro Thr Leu Glu His Thr 195 200
205 Leu Ile Asn Met Pro Asn Gly Phe Asn Ala Pro Ser Tyr
Leu Ser Thr 210 215 220
Thr Gln Pro Cys Leu Val Gln Gly Gly Asn Ile Val Ser Ala Asn Gly 225
230 235 240 Gly Asn Leu Phe
Tyr Gly Asn Asn His Gly Ile Phe Gly Gly Asn Leu 245
250 255 Ser Met Glu Gly His Glu Leu Tyr Val
Pro Pro Leu Glu Asn Val Ser 260 265
270 Ile Glu Tyr Gln Asn Val Glu Asn Gly Asn Phe Ser His His
Gln Asn 275 280 285
Asn Asn Asn Pro Asn Asn Met Thr Asn Leu Ile Asn Thr Ser His Asn 290
295 300 Phe Asn Thr Cys Ser
Asn Ile Lys Val Glu Asn Phe Gly Gly Ile Gly 305 310
315 320 Asn Tyr Trp Glu Gly Asp Asp Leu Lys Val
Gly Glu Trp Asp Leu Glu 325 330
335 Glu Leu Met Lys Asp Val Ser Pro Phe Pro Phe Leu Asp Phe Gln
Val 340 345 350 Glu
371002DNAGlycine maxGlyma15g03920.1 37atgaggaagc cagaggcgag taataataat
actaaaaaca acaacaacaa tagcaagaag 60cttagaaagg ggttgtggtc acctgaagaa
gatgacaagc tcatgaacta catgctaaac 120catggacaag ggtgttggag cgatgtggca
agaaatgctg gcctccaaag gtgtggcaaa 180agttgtcgcc ttcgatggat caattacttg
aggcctgatc ttaagagagg tgcattctca 240ccccaagaag aggaactcat catccacttc
cattcccttc ttggaaacag atggtctcaa 300atagcggcgc gtttgcctgg gcgaaccgac
aatgaaataa aaaacttttg gaattcgacg 360ataaagaaaa gactcaggaa tatgtcttcc
acgaccacca caacctcacc ctcaccctca 420tcaaatgcaa gcgagacctc aatatccgag
cctagtaata aagacctcaa catgggaggg 480tttatttcca cacaacataa tcaacacgca
ggctttgttc ctatgttcgg ttcatctcca 540tcaccatcaa taatgcaaac cggtacagtt
ttcaatacct tgattgacag attgcctatg 600ctggagcatg gactaaacat gccagcttct
ggagggtact tcgaaggcac aggtattcct 660tgcttttcgc aaagtgaagt taacaaatta
ggttcttgtt atttagaaaa cggagtattt 720gggagaagtg taaatatcgg ggtagaaggg
gatatgtttg ttcctcccct agagaatgct 780acatgcagca ggagagaaac aactaacagc
agttactttg acgatgacat aaatagtatt 840cttaataact gcaacattgg cataggtgaa
aataaggctc atgatggggt ggagaatttg 900tttcaacaag agttagccac tgccactgcc
acaggagaat gggactttga ggagttgatg 960aaattagatg tttcctcctt tccgtttctt
gatttttcat ac 100238334PRTGlycine maxGlyma15g03920.1
38Met Arg Lys Pro Glu Ala Ser Asn Asn Asn Thr Lys Asn Asn Asn Asn 1
5 10 15 Asn Ser Lys Lys
Leu Arg Lys Gly Leu Trp Ser Pro Glu Glu Asp Asp 20
25 30 Lys Leu Met Asn Tyr Met Leu Asn His
Gly Gln Gly Cys Trp Ser Asp 35 40
45 Val Ala Arg Asn Ala Gly Leu Gln Arg Cys Gly Lys Ser Cys
Arg Leu 50 55 60
Arg Trp Ile Asn Tyr Leu Arg Pro Asp Leu Lys Arg Gly Ala Phe Ser 65
70 75 80 Pro Gln Glu Glu Glu
Leu Ile Ile His Phe His Ser Leu Leu Gly Asn 85
90 95 Arg Trp Ser Gln Ile Ala Ala Arg Leu Pro
Gly Arg Thr Asp Asn Glu 100 105
110 Ile Lys Asn Phe Trp Asn Ser Thr Ile Lys Lys Arg Leu Arg Asn
Met 115 120 125 Ser
Ser Thr Thr Thr Thr Thr Ser Pro Ser Pro Ser Ser Asn Ala Ser 130
135 140 Glu Thr Ser Ile Ser Glu
Pro Ser Asn Lys Asp Leu Asn Met Gly Gly 145 150
155 160 Phe Ile Ser Thr Gln His Asn Gln His Ala Gly
Phe Val Pro Met Phe 165 170
175 Gly Ser Ser Pro Ser Pro Ser Ile Met Gln Thr Gly Thr Val Phe Asn
180 185 190 Thr Leu
Ile Asp Arg Leu Pro Met Leu Glu His Gly Leu Asn Met Pro 195
200 205 Ala Ser Gly Gly Tyr Phe Glu
Gly Thr Gly Ile Pro Cys Phe Ser Gln 210 215
220 Ser Glu Val Asn Lys Leu Gly Ser Cys Tyr Leu Glu
Asn Gly Val Phe 225 230 235
240 Gly Arg Ser Val Asn Ile Gly Val Glu Gly Asp Met Phe Val Pro Pro
245 250 255 Leu Glu Asn
Ala Thr Cys Ser Arg Arg Glu Thr Thr Asn Ser Ser Tyr 260
265 270 Phe Asp Asp Asp Ile Asn Ser Ile
Leu Asn Asn Cys Asn Ile Gly Ile 275 280
285 Gly Glu Asn Lys Ala His Asp Gly Val Glu Asn Leu Phe
Gln Gln Glu 290 295 300
Leu Ala Thr Ala Thr Ala Thr Gly Glu Trp Asp Phe Glu Glu Leu Met 305
310 315 320 Lys Leu Asp Val
Ser Ser Phe Pro Phe Leu Asp Phe Ser Tyr 325
330 39828DNAGlycine maxGlyma12g06180.1 39atgaggaagc
ccgaggtttc tggaaacaac aacaacaaca acaacattaa taacaagctt 60agaaaagggt
tgtggtcacc tgaagaagat gacaagctca tgaactacat gctaaacagt 120ggacaaggtt
gttggagcga tgtagccaga aatgctggcc ttcaaaggtg tggcaaaagt 180tgtcgccttc
gatggatcaa ctacttgagg cctgatctta aacgaggtgc attctcacaa 240caagaagagg
aactcatcat ccacttgcat tcccttctcg gaaacagatg gtctcaaata 300gcggcgcgct
taccagggag aacagacaat gaaattaaga atttttggaa ttcaacaata 360aagaaaagac
tcaagaacat gtcatccaac acctcaccaa atggaagcga gtcctcatat 420gagcctaata
acagagacct taacatggca gggtttacta cttctaatac ccaagatcaa 480caacatgctg
attttatgcc tatgttcaat tcatcatctc aatcaccctc catgcatgcc 540atggttctca
attccataat tgacaggttg cctatgctag agcatggact aaacatgcca 600tgttctgttg
acaacaaagg gatttatttg gaaaatggag gagtatttgg gagtgtaaat 660attggtgcag
aaggggatgt gtatgttccc cctctagaga gtgttagcac tacttctgac 720cataacctga
aaggctggtg gggtggagaa tttgtttcag gaagagttaa ccattggaga 780gtgggacttg
gaggagttaa tgaaagatgt ttcatccttt ccctttct
82840276PRTGlycine maxGlyma12g06180.1 40Met Arg Lys Pro Glu Val Ser Gly
Asn Asn Asn Asn Asn Asn Asn Ile 1 5 10
15 Asn Asn Lys Leu Arg Lys Gly Leu Trp Ser Pro Glu Glu
Asp Asp Lys 20 25 30
Leu Met Asn Tyr Met Leu Asn Ser Gly Gln Gly Cys Trp Ser Asp Val
35 40 45 Ala Arg Asn Ala
Gly Leu Gln Arg Cys Gly Lys Ser Cys Arg Leu Arg 50
55 60 Trp Ile Asn Tyr Leu Arg Pro Asp
Leu Lys Arg Gly Ala Phe Ser Gln 65 70
75 80 Gln Glu Glu Glu Leu Ile Ile His Leu His Ser Leu
Leu Gly Asn Arg 85 90
95 Trp Ser Gln Ile Ala Ala Arg Leu Pro Gly Arg Thr Asp Asn Glu Ile
100 105 110 Lys Asn Phe
Trp Asn Ser Thr Ile Lys Lys Arg Leu Lys Asn Met Ser 115
120 125 Ser Asn Thr Ser Pro Asn Gly Ser
Glu Ser Ser Tyr Glu Pro Asn Asn 130 135
140 Arg Asp Leu Asn Met Ala Gly Phe Thr Thr Ser Asn Thr
Gln Asp Gln 145 150 155
160 Gln His Ala Asp Phe Met Pro Met Phe Asn Ser Ser Ser Gln Ser Pro
165 170 175 Ser Met His Ala
Met Val Leu Asn Ser Ile Ile Asp Arg Leu Pro Met 180
185 190 Leu Glu His Gly Leu Asn Met Pro Cys
Ser Val Asp Asn Lys Gly Ile 195 200
205 Tyr Leu Glu Asn Gly Gly Val Phe Gly Ser Val Asn Ile Gly
Ala Glu 210 215 220
Gly Asp Val Tyr Val Pro Pro Leu Glu Ser Val Ser Thr Thr Ser Asp 225
230 235 240 His Asn Leu Lys Gly
Trp Trp Gly Gly Glu Phe Val Ser Gly Arg Val 245
250 255 Asn His Trp Arg Val Gly Leu Gly Gly Val
Asn Glu Arg Cys Phe Ile 260 265
270 Leu Ser Leu Ser 275 41887DNAGlycine
maxGlyma11g14200.1 41atgaggaagc ccgaggtttc tggaaaaaac aacaacatta
ataacaagct tagaaagggg 60ttgtggtcac ctgaagaaga tgacaagctc atgaactaca
tgctaaacag tggacaaggt 120tgttggagcg atgtagccag aaatgcgggc cttcaaaggt
gtggcaaaag ttgtcgcctt 180cgatggatca attacttgag gcctgatctt aaacgaggtg
cattctcacc acaagaagag 240gaaatcatca tccatttgca ttcccttctc ggaaacagat
ggtctcaaat agcagcgcgc 300ttaccaggga gaactgacaa tgaaattaag aacttttgga
attcaacaat aaagaaaaga 360ctcaagaact tgtcctccaa cacctcacca aatggaagcg
agtcatcata tgagcccaac 420aacaaagacc ttaacatggc agggtttact acttctaata
cccaacaaaa tcaacaacat 480gctgatttta tgcctatgtt gcctatgcta gagcatggac
taaacatgac aagttctggt 540ggattcttca acagcaaagg gccatgcttc tcatcatcac
aaagagtatt tgggagtgta 600aatattggtg cagaagggga tatgtatgtt cctcctctag
agagtgttag cactacttct 660gaccattata acctgaaatt ggagagtacg tgcaacacag
atactaacaa tagtaattac 720tttgatgaca taaacagtat catccttaat aactgcaaca
ttaataatag caacaatatt 780aagagagctg aaaatagggc tggtggggtg gagaatttgt
ttcaagaaga gttaaccatt 840ggagagtggg acttagagga gttgatgaaa gatgtttcat
cctttcc 88742296PRTGlycine
maxmisc_feature(296)..(296)Xaa can be any naturally occurring amino acid
42Met Arg Lys Pro Glu Val Ser Gly Lys Asn Asn Asn Ile Asn Asn Lys 1
5 10 15 Leu Arg Lys Gly
Leu Trp Ser Pro Glu Glu Asp Asp Lys Leu Met Asn 20
25 30 Tyr Met Leu Asn Ser Gly Gln Gly Cys
Trp Ser Asp Val Ala Arg Asn 35 40
45 Ala Gly Leu Gln Arg Cys Gly Lys Ser Cys Arg Leu Arg Trp
Ile Asn 50 55 60
Tyr Leu Arg Pro Asp Leu Lys Arg Gly Ala Phe Ser Pro Gln Glu Glu 65
70 75 80 Glu Ile Ile Ile His
Leu His Ser Leu Leu Gly Asn Arg Trp Ser Gln 85
90 95 Ile Ala Ala Arg Leu Pro Gly Arg Thr Asp
Asn Glu Ile Lys Asn Phe 100 105
110 Trp Asn Ser Thr Ile Lys Lys Arg Leu Lys Asn Leu Ser Ser Asn
Thr 115 120 125 Ser
Pro Asn Gly Ser Glu Ser Ser Tyr Glu Pro Asn Asn Lys Asp Leu 130
135 140 Asn Met Ala Gly Phe Thr
Thr Ser Asn Thr Gln Gln Asn Gln Gln His 145 150
155 160 Ala Asp Phe Met Pro Met Leu Pro Met Leu Glu
His Gly Leu Asn Met 165 170
175 Thr Ser Ser Gly Gly Phe Phe Asn Ser Lys Gly Pro Cys Phe Ser Ser
180 185 190 Ser Gln
Arg Val Phe Gly Ser Val Asn Ile Gly Ala Glu Gly Asp Met 195
200 205 Tyr Val Pro Pro Leu Glu Ser
Val Ser Thr Thr Ser Asp His Tyr Asn 210 215
220 Leu Lys Leu Glu Ser Thr Cys Asn Thr Asp Thr Asn
Asn Ser Asn Tyr 225 230 235
240 Phe Asp Asp Ile Asn Ser Ile Ile Leu Asn Asn Cys Asn Ile Asn Asn
245 250 255 Ser Asn Asn
Ile Lys Arg Ala Glu Asn Arg Ala Gly Gly Val Glu Asn 260
265 270 Leu Phe Gln Glu Glu Leu Thr Ile
Gly Glu Trp Asp Leu Glu Glu Leu 275 280
285 Met Lys Asp Val Ser Ser Phe Xaa 290
295 43840DNAArabidopsis thalianaAT5G12870.1 43atgaggaagc
cagaggtagc cattgcagct agtactcacc aagtaaagaa gatgaagaag 60ggactttggt
ctcctgagga agactcaaag ctgatgcaat acatgttaag caatggacaa 120ggatgttgga
gtgatgttgc gaaaaacgca ggacttcaaa gatgtggcaa aagctgccgt 180cttcgttgga
tcaactatct tcgtcctgac ctcaagcgtg gcgctttctc tcctcaagaa 240gaggatctca
tcattcgctt tcattccatc ctcggcaaca ggtggtctca gattgcagca 300cgattgcctg
gtcggaccga taacgagatc aagaatttct ggaactcaac aataaagaaa 360aggctaaaga
agatgtccga tacctccaac ttaatcaaca actcatcctc atcacccaac 420acagcaagcg
attcctcttc taattccgca tcttctttgg atattaaaga cattatagga 480agcttcatgt
ccttacaaga acaaggcttc gtcaaccctt ccttgaccca catacaaacc 540aacaatccat
ttccaacggg aaacatgatc agccacccgt gcaatgacga ttttacccct 600tatgtagatg
gtatctatgg agtaaacgca ggggtacaag gggaactcta cttcccacct 660ttggaatgtg
aagaaggtga ttggtacaat gcaaatataa acaaccactt agacgagttg 720aacactaatg
gatccggaaa cgcacctgag ggtatgagac cagtggaaga attttgggac 780cttgaccagt
tgatgaacac tgaggttcct tcgttttact tcaacttcaa acaaagcata
84044280PRTArabidopsis thalianaAT5G12870.1 44Met Arg Lys Pro Glu Val Ala
Ile Ala Ala Ser Thr His Gln Val Lys 1 5
10 15 Lys Met Lys Lys Gly Leu Trp Ser Pro Glu Glu
Asp Ser Lys Leu Met 20 25
30 Gln Tyr Met Leu Ser Asn Gly Gln Gly Cys Trp Ser Asp Val Ala
Lys 35 40 45 Asn
Ala Gly Leu Gln Arg Cys Gly Lys Ser Cys Arg Leu Arg Trp Ile 50
55 60 Asn Tyr Leu Arg Pro Asp
Leu Lys Arg Gly Ala Phe Ser Pro Gln Glu 65 70
75 80 Glu Asp Leu Ile Ile Arg Phe His Ser Ile Leu
Gly Asn Arg Trp Ser 85 90
95 Gln Ile Ala Ala Arg Leu Pro Gly Arg Thr Asp Asn Glu Ile Lys Asn
100 105 110 Phe Trp
Asn Ser Thr Ile Lys Lys Arg Leu Lys Lys Met Ser Asp Thr 115
120 125 Ser Asn Leu Ile Asn Asn Ser
Ser Ser Ser Pro Asn Thr Ala Ser Asp 130 135
140 Ser Ser Ser Asn Ser Ala Ser Ser Leu Asp Ile Lys
Asp Ile Ile Gly 145 150 155
160 Ser Phe Met Ser Leu Gln Glu Gln Gly Phe Val Asn Pro Ser Leu Thr
165 170 175 His Ile Gln
Thr Asn Asn Pro Phe Pro Thr Gly Asn Met Ile Ser His 180
185 190 Pro Cys Asn Asp Asp Phe Thr Pro
Tyr Val Asp Gly Ile Tyr Gly Val 195 200
205 Asn Ala Gly Val Gln Gly Glu Leu Tyr Phe Pro Pro Leu
Glu Cys Glu 210 215 220
Glu Gly Asp Trp Tyr Asn Ala Asn Ile Asn Asn His Leu Asp Glu Leu 225
230 235 240 Asn Thr Asn Gly
Ser Gly Asn Ala Pro Glu Gly Met Arg Pro Val Glu 245
250 255 Glu Phe Trp Asp Leu Asp Gln Leu Met
Asn Thr Glu Val Pro Ser Phe 260 265
270 Tyr Phe Asn Phe Lys Gln Ser Ile 275
280 451056DNASolanum lycopersicumSolyc01g087130.2.1 45atgaggaagc
cggaacacaa taatacaacg atgaaggaga aggagaagga gaaagtgaac 60aagttaggga
aattaagaaa aggtttatgg tcaccagaag aagatgaaaa gttgatgagt 120tacatgttaa
gaaatggtca aggatgttgg agtgacattg ctagaaatgc tggattacaa 180agatgtggta
agagttgtcg tcttagatgg attaattact tgaggcctga ccttaaacgc 240ggcgcctttt
ctcttcaaga agaagaactc attgttcatt tgcattctat tctgggaaat 300aggtggtcac
aaattgcggc tcgtctacct gggaggacag ataatgaaat caagaatttc 360tggaattcca
cgataaaaaa gagatcaaaa aacaacaaca acagcacgcc atcgcccaac 420acgagtgatt
cttccattgg aggaatattt ccaatgcaag ggcacgatgt aaataatgtt 480atggcaacgt
tatgcatgga taattcatcg tctactactt caggatcatc catgcaagcc 540atggtacctt
tcaacccttt ccctcagctt gatagtacaa gctacgatat atctggatta 600gtcgggccag
tgaatttagg tcaatttggt tgtagtggag gtgatggtgg atttttggac 660tatggagttg
tggaaactta tagtatgatg ggtttaggaa gtgatgaatt ttcaatacct 720tctttagagg
gtgttcataa taagagtact actactatgg gagagagtaa taataatagt 780aatgttgatt
ttagtagtaa taatattgtt agtggtgcta atgactatga ttccatgatt 840gagaagaaaa
ataatacaaa cgttaacaac aacaacaacc aacacttgat gaatatgagt 900ggaattagtg
atcaaagcct aaaggttgaa gactatatgg ttggttttgg gaatcatcat 960cattggcatg
gagaaagctt aagaattgga gaatttgatt gggaaggttt gttggcaaat 1020gtttcctctt
taccttacct tgattttcaa gttgaa
105646352PRTSolanum lycopersicumSolyc01g087130.2.1 46Met Arg Lys Pro Glu
His Asn Asn Thr Thr Met Lys Glu Lys Glu Lys 1 5
10 15 Glu Lys Val Asn Lys Leu Gly Lys Leu Arg
Lys Gly Leu Trp Ser Pro 20 25
30 Glu Glu Asp Glu Lys Leu Met Ser Tyr Met Leu Arg Asn Gly Gln
Gly 35 40 45 Cys
Trp Ser Asp Ile Ala Arg Asn Ala Gly Leu Gln Arg Cys Gly Lys 50
55 60 Ser Cys Arg Leu Arg Trp
Ile Asn Tyr Leu Arg Pro Asp Leu Lys Arg 65 70
75 80 Gly Ala Phe Ser Leu Gln Glu Glu Glu Leu Ile
Val His Leu His Ser 85 90
95 Ile Leu Gly Asn Arg Trp Ser Gln Ile Ala Ala Arg Leu Pro Gly Arg
100 105 110 Thr Asp
Asn Glu Ile Lys Asn Phe Trp Asn Ser Thr Ile Lys Lys Arg 115
120 125 Ser Lys Asn Asn Asn Asn Ser
Thr Pro Ser Pro Asn Thr Ser Asp Ser 130 135
140 Ser Ile Gly Gly Ile Phe Pro Met Gln Gly His Asp
Val Asn Asn Val 145 150 155
160 Met Ala Thr Leu Cys Met Asp Asn Ser Ser Ser Thr Thr Ser Gly Ser
165 170 175 Ser Met Gln
Ala Met Val Pro Phe Asn Pro Phe Pro Gln Leu Asp Ser 180
185 190 Thr Ser Tyr Asp Ile Ser Gly Leu
Val Gly Pro Val Asn Leu Gly Gln 195 200
205 Phe Gly Cys Ser Gly Gly Asp Gly Gly Phe Leu Asp Tyr
Gly Val Val 210 215 220
Glu Thr Tyr Ser Met Met Gly Leu Gly Ser Asp Glu Phe Ser Ile Pro 225
230 235 240 Ser Leu Glu Gly
Val His Asn Lys Ser Thr Thr Thr Met Gly Glu Ser 245
250 255 Asn Asn Asn Ser Asn Val Asp Phe Ser
Ser Asn Asn Ile Val Ser Gly 260 265
270 Ala Asn Asp Tyr Asp Ser Met Ile Glu Lys Lys Asn Asn Thr
Asn Val 275 280 285
Asn Asn Asn Asn Asn Gln His Leu Met Asn Met Ser Gly Ile Ser Asp 290
295 300 Gln Ser Leu Lys Val
Glu Asp Tyr Met Val Gly Phe Gly Asn His His 305 310
315 320 His Trp His Gly Glu Ser Leu Arg Ile Gly
Glu Phe Asp Trp Glu Gly 325 330
335 Leu Leu Ala Asn Val Ser Ser Leu Pro Tyr Leu Asp Phe Gln Val
Glu 340 345 350
471008DNAGlycine maxGlyma19g05080.1 47atgaggaaac ctgatatgat gggaaaagac
aaaatcaaca acaacattaa gagcaagcta 60aggaagggtt tgtggtcacc tgaggaagat
gagaagctcc taaggtatat gatcactaag 120ggacaagggt gttggagtga cattgctagg
aatgctggtc ttcaaaggtg cggcaaaagt 180tgccgtcttc gttggattaa ctacttgaga
cctgatctca aacgtggtgc attttcacct 240caagaggaag aagtcatcat tcacttgcac
tccattcttg gcaacagatg gtctcaaatt 300gccgcacgtc tccctggtcg cacagacaat
gagatcaaga atttctggaa ctccacactg 360aagaaaaggt tgaaaatgaa caacaataac
tccactttat caccaaacaa tagtgactca 420tcagggccta aagatgtcaa tgtcatgggt
ggaatcatgt ccatgaacga gcatgacctc 480atgaccatgt gcatggactc ctcctcatca
acatcatcat catcatgcat gcaatccatg 540catgccacca acatggtact aactaacccc
tttcccttgt tgcccaacaa ccgttatgac 600atgatgaccg gtgcaaccgg tttccttgac
aacatggctg ctgcatgctt aacccaagtt 660ggcatggtag atcatgatca tggggttgtt
catgggacat tggagcctaa taaaacgcgt 720ttaggaagcg acttttccct tcctccacta
gaaagtagaa gcattgagga caatagtagt 780accccaattg atcatgtgaa aagccataac
aacaacaacc acttcaagaa tagttgcttc 840aataacactg atcattacca tcatattcaa
agctccaaca acgtagttgt agaggatttg 900tttgggtttg gaaatcatgg gcaaggggaa
aactttagaa tgggagaatg ggaccttgag 960ggcttgatgc aagacatttc ctattttcct
tcccttgatt tccaagtt 100848336PRTGlycine maxGlyma19g05080.1
48Met Arg Lys Pro Asp Met Met Gly Lys Asp Lys Ile Asn Asn Asn Ile 1
5 10 15 Lys Ser Lys Leu
Arg Lys Gly Leu Trp Ser Pro Glu Glu Asp Glu Lys 20
25 30 Leu Leu Arg Tyr Met Ile Thr Lys Gly
Gln Gly Cys Trp Ser Asp Ile 35 40
45 Ala Arg Asn Ala Gly Leu Gln Arg Cys Gly Lys Ser Cys Arg
Leu Arg 50 55 60
Trp Ile Asn Tyr Leu Arg Pro Asp Leu Lys Arg Gly Ala Phe Ser Pro 65
70 75 80 Gln Glu Glu Glu Val
Ile Ile His Leu His Ser Ile Leu Gly Asn Arg 85
90 95 Trp Ser Gln Ile Ala Ala Arg Leu Pro Gly
Arg Thr Asp Asn Glu Ile 100 105
110 Lys Asn Phe Trp Asn Ser Thr Leu Lys Lys Arg Leu Lys Met Asn
Asn 115 120 125 Asn
Asn Ser Thr Leu Ser Pro Asn Asn Ser Asp Ser Ser Gly Pro Lys 130
135 140 Asp Val Asn Val Met Gly
Gly Ile Met Ser Met Asn Glu His Asp Leu 145 150
155 160 Met Thr Met Cys Met Asp Ser Ser Ser Ser Thr
Ser Ser Ser Ser Cys 165 170
175 Met Gln Ser Met His Ala Thr Asn Met Val Leu Thr Asn Pro Phe Pro
180 185 190 Leu Leu
Pro Asn Asn Arg Tyr Asp Met Met Thr Gly Ala Thr Gly Phe 195
200 205 Leu Asp Asn Met Ala Ala Ala
Cys Leu Thr Gln Val Gly Met Val Asp 210 215
220 His Asp His Gly Val Val His Gly Thr Leu Glu Pro
Asn Lys Thr Arg 225 230 235
240 Leu Gly Ser Asp Phe Ser Leu Pro Pro Leu Glu Ser Arg Ser Ile Glu
245 250 255 Asp Asn Ser
Ser Thr Pro Ile Asp His Val Lys Ser His Asn Asn Asn 260
265 270 Asn His Phe Lys Asn Ser Cys Phe
Asn Asn Thr Asp His Tyr His His 275 280
285 Ile Gln Ser Ser Asn Asn Val Val Val Glu Asp Leu Phe
Gly Phe Gly 290 295 300
Asn His Gly Gln Gly Glu Asn Phe Arg Met Gly Glu Trp Asp Leu Glu 305
310 315 320 Gly Leu Met Gln
Asp Ile Ser Tyr Phe Pro Ser Leu Asp Phe Gln Val 325
330 335 49933DNAGlycine maxGlyma13g27310.1
49atgaggaaac ctgatctgat ggccaacaag gacaaagtga acaacaacat aaagagcaag
60ttgagaaaag ggttgtggtc accagatgaa gatgagaggc tcataaggta catgctcaca
120aatggacaag ggtgttggag tgacattgct aggaatgctg gtcttcaaag gtgtggcaaa
180agttgccgtc ttcgttggat caattacttg agacctgacc tcaagcgtgg tgcattttcg
240ccccaagagg aagatctcat cgttcatttg cactccattc ttggcaatag gtggtctcag
300attgcagcac atctccctgg ccgcacagac aatgagatta agaatttctg gaactccaca
360ttgaagaaaa ggttgaaagc aaacacttct actccctcac taaacaacag cacaggctca
420tcagagtcta ataaggatgt tttgagtggg atcatgccct ttagtgaaca tgacatcatg
480accatgtgca tggattcctc ttcttccata tcatccatgc aagcaacggt tttgcctgac
540caatttgacc ctttttccat gttggcaaat aatcagtgtg acatgactaa tgtttcagca
600gattttccca acttgactca aattggcatg gtagaggggc atgaagggaa ttatgggata
660ttggagccaa ataaaatggg gttaggaaga gatttctccc ttccttcact agaaagtaga
720agcattgaaa gcaatagtgt cccaattgat gtgaaaagcc ataacaacca cttcaattat
780ggttccttca atcacactga taaaattcag ggctccaaag tagaggactt aattgagttt
840ggaaatcatg gccaagggga ggatttaaaa atgggagagt gggatttgga gaatttgatg
900caagacataa cctcttttcc tttccttgag ttt
93350311PRTGlycine maxGlyma13g27310.1 50Met Arg Lys Pro Asp Leu Met Ala
Asn Lys Asp Lys Val Asn Asn Asn 1 5 10
15 Ile Lys Ser Lys Leu Arg Lys Gly Leu Trp Ser Pro Asp
Glu Asp Glu 20 25 30
Arg Leu Ile Arg Tyr Met Leu Thr Asn Gly Gln Gly Cys Trp Ser Asp
35 40 45 Ile Ala Arg Asn
Ala Gly Leu Gln Arg Cys Gly Lys Ser Cys Arg Leu 50
55 60 Arg Trp Ile Asn Tyr Leu Arg Pro
Asp Leu Lys Arg Gly Ala Phe Ser 65 70
75 80 Pro Gln Glu Glu Asp Leu Ile Val His Leu His Ser
Ile Leu Gly Asn 85 90
95 Arg Trp Ser Gln Ile Ala Ala His Leu Pro Gly Arg Thr Asp Asn Glu
100 105 110 Ile Lys Asn
Phe Trp Asn Ser Thr Leu Lys Lys Arg Leu Lys Ala Asn 115
120 125 Thr Ser Thr Pro Ser Leu Asn Asn
Ser Thr Gly Ser Ser Glu Ser Asn 130 135
140 Lys Asp Val Leu Ser Gly Ile Met Pro Phe Ser Glu His
Asp Ile Met 145 150 155
160 Thr Met Cys Met Asp Ser Ser Ser Ser Ile Ser Ser Met Gln Ala Thr
165 170 175 Val Leu Pro Asp
Gln Phe Asp Pro Phe Ser Met Leu Ala Asn Asn Gln 180
185 190 Cys Asp Met Thr Asn Val Ser Ala Asp
Phe Pro Asn Leu Thr Gln Ile 195 200
205 Gly Met Val Glu Gly His Glu Gly Asn Tyr Gly Ile Leu Glu
Pro Asn 210 215 220
Lys Met Gly Leu Gly Arg Asp Phe Ser Leu Pro Ser Leu Glu Ser Arg 225
230 235 240 Ser Ile Glu Ser Asn
Ser Val Pro Ile Asp Val Lys Ser His Asn Asn 245
250 255 His Phe Asn Tyr Gly Ser Phe Asn His Thr
Asp Lys Ile Gln Gly Ser 260 265
270 Lys Val Glu Asp Leu Ile Glu Phe Gly Asn His Gly Gln Gly Glu
Asp 275 280 285 Leu
Lys Met Gly Glu Trp Asp Leu Glu Asn Leu Met Gln Asp Ile Thr 290
295 300 Ser Phe Pro Phe Leu Glu
Phe 305 310 51945DNAGlycine maxGlyma12g36630.1
51atgaggaaac ctgatctgat ggccaacaag gacaaaatga acaacattaa gagcaagttg
60agaaaagggt tgtggtcacc agatgaagat gagaggctcg taaggtacat gctgacaaat
120ggacaagggt gttggagtga cattgctagg aatgctggtc ttcaaaggtg tggcaaaagt
180tgccgtcttc gttggatcaa ttacttgaga cctgacctca agcgtggtgc attctcacct
240caagaggaag atctcatcgt tcatttgcac tccattcttg gcaataggtg gtctcagatt
300gcagcgcgtc tccctggccg cacagacaat gagattaaga atttctggaa ctccacattg
360aagaaaaggt tgaaaactaa cacttccact ccctcactaa acaacagcac tggctcatca
420gagtctaata aggatgtttt gagtgggatc atgcccttta atgaacatga catcatgacc
480atgtgcatgg attcctcttc gtccatatca tccatgcaag caatggtttt gcctgaccaa
540tttgaccctt ttttcatgtt ggcaaataat cagtgtgaca tgactaatgt ttcatctgac
600ttttccaaca tgcctgctgc atgcttgact caaattggca tggtagatgg gcatcaaggg
660aattatggga tattggagcc aaataaaatg gggtcaggaa tagacttctc ccttccttca
720ctagaaagta gaagcattga aagcaatagt gtcccaattg atgtgaaaag ccataacaac
780cacttcaatt atggttcctt caagaacact gataagattc agggctccaa agtggaggac
840ttgatcgggt ttggaaatca tggccaaggg gaaaatttaa aaatgggaga gtgggatttg
900gagaatttaa tgcaagacat aacctctttt cctttccttg atttt
94552315PRTGlycine maxGlyma12g36630.1 52Met Arg Lys Pro Asp Leu Met Ala
Asn Lys Asp Lys Met Asn Asn Ile 1 5 10
15 Lys Ser Lys Leu Arg Lys Gly Leu Trp Ser Pro Asp Glu
Asp Glu Arg 20 25 30
Leu Val Arg Tyr Met Leu Thr Asn Gly Gln Gly Cys Trp Ser Asp Ile
35 40 45 Ala Arg Asn Ala
Gly Leu Gln Arg Cys Gly Lys Ser Cys Arg Leu Arg 50
55 60 Trp Ile Asn Tyr Leu Arg Pro Asp
Leu Lys Arg Gly Ala Phe Ser Pro 65 70
75 80 Gln Glu Glu Asp Leu Ile Val His Leu His Ser Ile
Leu Gly Asn Arg 85 90
95 Trp Ser Gln Ile Ala Ala Arg Leu Pro Gly Arg Thr Asp Asn Glu Ile
100 105 110 Lys Asn Phe
Trp Asn Ser Thr Leu Lys Lys Arg Leu Lys Thr Asn Thr 115
120 125 Ser Thr Pro Ser Leu Asn Asn Ser
Thr Gly Ser Ser Glu Ser Asn Lys 130 135
140 Asp Val Leu Ser Gly Ile Met Pro Phe Asn Glu His Asp
Ile Met Thr 145 150 155
160 Met Cys Met Asp Ser Ser Ser Ser Ile Ser Ser Met Gln Ala Met Val
165 170 175 Leu Pro Asp Gln
Phe Asp Pro Phe Phe Met Leu Ala Asn Asn Gln Cys 180
185 190 Asp Met Thr Asn Val Ser Ser Asp Phe
Ser Asn Met Pro Ala Ala Cys 195 200
205 Leu Thr Gln Ile Gly Met Val Asp Gly His Gln Gly Asn Tyr
Gly Ile 210 215 220
Leu Glu Pro Asn Lys Met Gly Ser Gly Ile Asp Phe Ser Leu Pro Ser 225
230 235 240 Leu Glu Ser Arg Ser
Ile Glu Ser Asn Ser Val Pro Ile Asp Val Lys 245
250 255 Ser His Asn Asn His Phe Asn Tyr Gly Ser
Phe Lys Asn Thr Asp Lys 260 265
270 Ile Gln Gly Ser Lys Val Glu Asp Leu Ile Gly Phe Gly Asn His
Gly 275 280 285 Gln
Gly Glu Asn Leu Lys Met Gly Glu Trp Asp Leu Glu Asn Leu Met 290
295 300 Gln Asp Ile Thr Ser Phe
Pro Phe Leu Asp Phe 305 310 315
531029DNAArabidopsis thalianaAT3G08500.1 53atgatgatga ggaaaccgga
cattactacg atcagagaca aaggcaagcc aaatcatgca 60tgtggtggta ataacaacaa
accgaagcta agaaaaggac tttggtcgcc tgatgaagat 120gagaagctga taagatacat
gttgactaat ggacaaggat gttggagtga catcgctaga 180aatgctggtc ttttacgttg
tggtaaaagt tgtcgccttc gctggatcaa ttacttgagg 240cctgatctta aacgtggatc
cttctctcct caggaggagg atctcatctt ccatttgcat 300tccattcttg gtaacaggtg
gtctcaaata gctactcggc ttccaggtag aacagacaac 360gagatcaaaa acttttggaa
ctcgacattg aagaagcggc ttaagaacaa cagcaacaac 420aatacttcat caggatcatc
acctaacaat agtaatagta attccttgga cccaagagat 480caacatgtgg atatgggagg
caactcaact tcattgatgg atgactatca tcatgatgaa 540aacatgatga cagtggggaa
caccatgcgc atggactctt cctccccatt caatgttgga 600ccaatggtta atagtgtggg
cttaaaccaa ctttatgatc ccttgatgat atcagtgccg 660gataacggat atcaccaaat
gggaaacaca gtgaatgtgt tcagcgttaa tggtttagga 720gattatggaa acacaattct
tgatccaatt agcaagagag tatcagtaga aggtgatgat 780tggttcattc ccccctcgga
gaataccaac gtcattgctt gtagtacaag caacaaccta 840aacttacagg cccttgatcc
ttgcttcaat agcaaaaatc tttgtcattc agaaagcttc 900aaggtaggga atgtgttggg
gatagagaat ggttcttggg aaatagaaaa ccctaaaatc 960ggagattggg atttggatgg
tctcatcgat aacaactctt cttttccctt ccttgatttc 1020caagtcgat
102954343PRTArabidopsis
thalianaAT3G08500.1 54Met Met Met Arg Lys Pro Asp Ile Thr Thr Ile Arg Asp
Lys Gly Lys 1 5 10 15
Pro Asn His Ala Cys Gly Gly Asn Asn Asn Lys Pro Lys Leu Arg Lys
20 25 30 Gly Leu Trp Ser
Pro Asp Glu Asp Glu Lys Leu Ile Arg Tyr Met Leu 35
40 45 Thr Asn Gly Gln Gly Cys Trp Ser Asp
Ile Ala Arg Asn Ala Gly Leu 50 55
60 Leu Arg Cys Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn
Tyr Leu Arg 65 70 75
80 Pro Asp Leu Lys Arg Gly Ser Phe Ser Pro Gln Glu Glu Asp Leu Ile
85 90 95 Phe His Leu His
Ser Ile Leu Gly Asn Arg Trp Ser Gln Ile Ala Thr 100
105 110 Arg Leu Pro Gly Arg Thr Asp Asn Glu
Ile Lys Asn Phe Trp Asn Ser 115 120
125 Thr Leu Lys Lys Arg Leu Lys Asn Asn Ser Asn Asn Asn Thr
Ser Ser 130 135 140
Gly Ser Ser Pro Asn Asn Ser Asn Ser Asn Ser Leu Asp Pro Arg Asp 145
150 155 160 Gln His Val Asp Met
Gly Gly Asn Ser Thr Ser Leu Met Asp Asp Tyr 165
170 175 His His Asp Glu Asn Met Met Thr Val Gly
Asn Thr Met Arg Met Asp 180 185
190 Ser Ser Ser Pro Phe Asn Val Gly Pro Met Val Asn Ser Val Gly
Leu 195 200 205 Asn
Gln Leu Tyr Asp Pro Leu Met Ile Ser Val Pro Asp Asn Gly Tyr 210
215 220 His Gln Met Gly Asn Thr
Val Asn Val Phe Ser Val Asn Gly Leu Gly 225 230
235 240 Asp Tyr Gly Asn Thr Ile Leu Asp Pro Ile Ser
Lys Arg Val Ser Val 245 250
255 Glu Gly Asp Asp Trp Phe Ile Pro Pro Ser Glu Asn Thr Asn Val Ile
260 265 270 Ala Cys
Ser Thr Ser Asn Asn Leu Asn Leu Gln Ala Leu Asp Pro Cys 275
280 285 Phe Asn Ser Lys Asn Leu Cys
His Ser Glu Ser Phe Lys Val Gly Asn 290 295
300 Val Leu Gly Ile Glu Asn Gly Ser Trp Glu Ile Glu
Asn Pro Lys Ile 305 310 315
320 Gly Asp Trp Asp Leu Asp Gly Leu Ile Asp Asn Asn Ser Ser Phe Pro
325 330 335 Phe Leu Asp
Phe Gln Val Asp 340 551215DNAZea
maysGRMZM2G052606_T01 55atgaggaaac cggagtgccc agcggcgaac agcagcaatg
cgggggcggc ggccgcgaag 60ctgcggaagg ggctgtggtc gccggaggag gacgagaggc
tggtggcgta catgctgcgg 120agtggacagg gttcttggag cgatgtggcc cggaacgccg
ggttgcagcg gtgcggcaag 180agctgccgcc tccggtggat caactacctc cggccggacc
tcaagcgcgg cgccttctcg 240ccgcaggagg aggagctcat cgtcagcctc cacgccatcc
tgggaaacag gtggtctcag 300attgctgccc ggttgccggg gcgcaccgac aacgagatca
agaacttctg gaactccacc 360atcaagaagc ggctcaagaa cagctcggca gcttcgtcac
cagcagctac ggactgcgcg 420tcgccggagc ctaataacaa ggtcgccgcc gccggtagct
gcccggatct ttccgtccta 480gatcatcagg acggtggcca ccaccacgca atgacgacga
cgactgcagg tttgtggatg 540gtggactcat cctcctcttg tacctcgtcg acctcgccaa
tgcatcagtt tcagaggccg 600acgacgacga tggcagcggc cgtggccagc gggagctatg
gaggtctcgt ccccttccct 660gaccaggtcc gtggtgttgt ggccgacacg ggagggttct
ttcatggcca cgcggcgcca 720gcgttcaagc accaagttgc cgcattgcac ggtggtggtt
attactacgg cagcgctcct 780cgtcaccatg gaatgacgac gacgacgacg acggtggcat
tggaaggaag cggtggatgc 840ttcatatctg gcgaaggcat gcttggtgtg ccccctctgc
tgttagagcc catgtcagca 900gcgctagagc aagaccaagg ccagaccttg atggcatcaa
gtggtaacaa caaccctaaa 960aacaacagca gcagcaacac tactgatact acgactacca
cgacactgag caacaatgag 1020agcaacgtca cagacaccac caccaaggac aacaccacca
acaccatcag ccaagtgaac 1080agtggcagca ataatgtcta ctgggagggg gcccgccagc
agtacatgag caggaatgtc 1140atgcatgggg agtgggacct ggaggagctg atgaaagatg
tgtcatcctt gccttttctt 1200gatttccaag ttgaa
121556405PRTZea maysGRMZM2G052606_T01 56Met Arg Lys
Pro Glu Cys Pro Ala Ala Asn Ser Ser Asn Ala Gly Ala 1 5
10 15 Ala Ala Ala Lys Leu Arg Lys Gly
Leu Trp Ser Pro Glu Glu Asp Glu 20 25
30 Arg Leu Val Ala Tyr Met Leu Arg Ser Gly Gln Gly Ser
Trp Ser Asp 35 40 45
Val Ala Arg Asn Ala Gly Leu Gln Arg Cys Gly Lys Ser Cys Arg Leu 50
55 60 Arg Trp Ile Asn
Tyr Leu Arg Pro Asp Leu Lys Arg Gly Ala Phe Ser 65 70
75 80 Pro Gln Glu Glu Glu Leu Ile Val Ser
Leu His Ala Ile Leu Gly Asn 85 90
95 Arg Trp Ser Gln Ile Ala Ala Arg Leu Pro Gly Arg Thr Asp
Asn Glu 100 105 110
Ile Lys Asn Phe Trp Asn Ser Thr Ile Lys Lys Arg Leu Lys Asn Ser
115 120 125 Ser Ala Ala Ser
Ser Pro Ala Ala Thr Asp Cys Ala Ser Pro Glu Pro 130
135 140 Asn Asn Lys Val Ala Ala Ala Gly
Ser Cys Pro Asp Leu Ser Val Leu 145 150
155 160 Asp His Gln Asp Gly Gly His His His Ala Met Thr
Thr Thr Thr Ala 165 170
175 Gly Leu Trp Met Val Asp Ser Ser Ser Ser Cys Thr Ser Ser Thr Ser
180 185 190 Pro Met His
Gln Phe Gln Arg Pro Thr Thr Thr Met Ala Ala Ala Val 195
200 205 Ala Ser Gly Ser Tyr Gly Gly Leu
Val Pro Phe Pro Asp Gln Val Arg 210 215
220 Gly Val Val Ala Asp Thr Gly Gly Phe Phe His Gly His
Ala Ala Pro 225 230 235
240 Ala Phe Lys His Gln Val Ala Ala Leu His Gly Gly Gly Tyr Tyr Tyr
245 250 255 Gly Ser Ala Pro
Arg His His Gly Met Thr Thr Thr Thr Thr Thr Val 260
265 270 Ala Leu Glu Gly Ser Gly Gly Cys Phe
Ile Ser Gly Glu Gly Met Leu 275 280
285 Gly Val Pro Pro Leu Leu Leu Glu Pro Met Ser Ala Ala Leu
Glu Gln 290 295 300
Asp Gln Gly Gln Thr Leu Met Ala Ser Ser Gly Asn Asn Asn Pro Lys 305
310 315 320 Asn Asn Ser Ser Ser
Asn Thr Thr Asp Thr Thr Thr Thr Thr Thr Leu 325
330 335 Ser Asn Asn Glu Ser Asn Val Thr Asp Thr
Thr Thr Lys Asp Asn Thr 340 345
350 Thr Asn Thr Ile Ser Gln Val Asn Ser Gly Ser Asn Asn Val Tyr
Trp 355 360 365 Glu
Gly Ala Arg Gln Gln Tyr Met Ser Arg Asn Val Met His Gly Glu 370
375 380 Trp Asp Leu Glu Glu Leu
Met Lys Asp Val Ser Ser Leu Pro Phe Leu 385 390
395 400 Asp Phe Gln Val Glu 405
571236DNASetaria italicaSi024786m 57atgaggaagc cggagggccc agcggcgagc
ggcggctgca atggcggtgc ggcggcggcg 60gcgaagctgc ggaaggggct gtggtcgccg
gaggaggacg agaagctggt ggcctacatg 120ctgcggagcg ggcaggggtc gtggagcgac
gtggcccgga acgccggcct gcaacgctgc 180ggcaagagct gccgcctccg gtggatcaac
tacctccggc cggacctcaa gcgcggcgcc 240ttctcgccgc aggaggagga cctcatcgtc
agcctccacg ccatcctcgg caacaggtgg 300tctcagatcg ctgcccggct gccggggcgc
accgacaacg agatcaagaa cttctggaac 360tccaccatca agaagcggct caagaacagc
tcctcggcct cgtcgccggc ggccaccgac 420tgcgcgtcgc cgacggagcc tagcagcaag
gtcgccggca tcgacatcag cggcgccacc 480agctgcccgg acctcgccgg cctggaccat
catcatcagg acggcggcca ccaccacgcg 540atgatgacga cgggcttgtg gatggtggac
tcgtcctcct ccacttcctc atcgacctcg 600ccgatgcaga gccggccgcc gccgtcggcc
attgcagcgg cggtggcccg gagctacggc 660ggcctcctcc ccctccccga ccagctccgc
ggcggcacgg cggccgacac gtcgccggca 720gggttcttcc acggccacgc ggcgccgttc
aaacagcaag cagcagttgc ctcattgcat 780ggcggttact atggaatggg cagtcctcat
caccatggga tgatggcaat ggagggagga 840ggagggtgct tcatgagagg agaaggcctc
tttggtgtgg cccctctgct ggatgccatg 900tcagcacaag accaagacca ggcaggccag
gccctaatag catcaagtgg tggtaacaac 960aaccctaaaa acaacagcag caacaacact
accgagacta caacaacagt gagtaacaat 1020gagagcaaca tcacagacaa caacaccacc
aacaccaagg acaacaacat caacgccatg 1080agcctagtga acagcggcag cagcaatgtg
gctgctgtct actgggaggg ggcccaccag 1140cagtacatga gcaggaatgt catgcatggg
gagtgggacc tggaggagct gatgaaagat 1200gtgtcatcct tgcctttcct tgatttccaa
gtcgaa 123658412PRTSetaria italicaSi024786m
58Met Arg Lys Pro Glu Gly Pro Ala Ala Ser Gly Gly Cys Asn Gly Gly 1
5 10 15 Ala Ala Ala Ala
Ala Lys Leu Arg Lys Gly Leu Trp Ser Pro Glu Glu 20
25 30 Asp Glu Lys Leu Val Ala Tyr Met Leu
Arg Ser Gly Gln Gly Ser Trp 35 40
45 Ser Asp Val Ala Arg Asn Ala Gly Leu Gln Arg Cys Gly Lys
Ser Cys 50 55 60
Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro Asp Leu Lys Arg Gly Ala 65
70 75 80 Phe Ser Pro Gln Glu
Glu Asp Leu Ile Val Ser Leu His Ala Ile Leu 85
90 95 Gly Asn Arg Trp Ser Gln Ile Ala Ala Arg
Leu Pro Gly Arg Thr Asp 100 105
110 Asn Glu Ile Lys Asn Phe Trp Asn Ser Thr Ile Lys Lys Arg Leu
Lys 115 120 125 Asn
Ser Ser Ser Ala Ser Ser Pro Ala Ala Thr Asp Cys Ala Ser Pro 130
135 140 Thr Glu Pro Ser Ser Lys
Val Ala Gly Ile Asp Ile Ser Gly Ala Thr 145 150
155 160 Ser Cys Pro Asp Leu Ala Gly Leu Asp His His
His Gln Asp Gly Gly 165 170
175 His His His Ala Met Met Thr Thr Gly Leu Trp Met Val Asp Ser Ser
180 185 190 Ser Ser
Thr Ser Ser Ser Thr Ser Pro Met Gln Ser Arg Pro Pro Pro 195
200 205 Ser Ala Ile Ala Ala Ala Val
Ala Arg Ser Tyr Gly Gly Leu Leu Pro 210 215
220 Leu Pro Asp Gln Leu Arg Gly Gly Thr Ala Ala Asp
Thr Ser Pro Ala 225 230 235
240 Gly Phe Phe His Gly His Ala Ala Pro Phe Lys Gln Gln Ala Ala Val
245 250 255 Ala Ser Leu
His Gly Gly Tyr Tyr Gly Met Gly Ser Pro His His His 260
265 270 Gly Met Met Ala Met Glu Gly Gly
Gly Gly Cys Phe Met Arg Gly Glu 275 280
285 Gly Leu Phe Gly Val Ala Pro Leu Leu Asp Ala Met Ser
Ala Gln Asp 290 295 300
Gln Asp Gln Ala Gly Gln Ala Leu Ile Ala Ser Ser Gly Gly Asn Asn 305
310 315 320 Asn Pro Lys Asn
Asn Ser Ser Asn Asn Thr Thr Glu Thr Thr Thr Thr 325
330 335 Val Ser Asn Asn Glu Ser Asn Ile Thr
Asp Asn Asn Thr Thr Asn Thr 340 345
350 Lys Asp Asn Asn Ile Asn Ala Met Ser Leu Val Asn Ser Gly
Ser Ser 355 360 365
Asn Val Ala Ala Val Tyr Trp Glu Gly Ala His Gln Gln Tyr Met Ser 370
375 380 Arg Asn Val Met His
Gly Glu Trp Asp Leu Glu Glu Leu Met Lys Asp 385 390
395 400 Val Ser Ser Leu Pro Phe Leu Asp Phe Gln
Val Glu 405 410 591227DNAOryza
sativaLOC_Os12g33070.1 59atgaggaagc cggattgcgg cggcggggga ggggcggcga
agggcggcgg cgttctgggt 60gtggcgggag ggaacaatgc ggcggtggtg ggggggaagg
ttcggaaggg gctgtggtcg 120ccggaggagg acgagaagct ggtggcgtac atgctgcgga
gcgggcaggg gtcgtggagc 180gacgtggcga ggaacgccgg gctgcagcgc tgcggcaaga
gctgccgcct ccggtggatc 240aactacctcc gccccgacct caagcgcggc gccttctcgc
cgcaggagga ggacctcatc 300gtcaacctcc acgccatcct cggcaacagg tggtctcaga
tcgctgcccg gctaccgggg 360cgcaccgaca acgagatcaa gaacttctgg aactccacca
tcaagaagcg cctcaagatc 420tcctcctcct cggcgtctcc ggccaccacc accgactgcg
cctccccgcc ggagcacaag 480ctcggcgccg tcgtcgacct cgccggcggc ggcggcgcca
cggacgacgt cgttgtcggg 540acagctaatg ctgccatgaa gagcatgtgg gtggattcct
cgtcgtcgtc gtcgtcgtct 600tcctcgtcga tgcagagccg gccgtcgata atggcggcgg
cggcggcggg gaggagctac 660ggcggcctcc tcccactccc cgaccaggtc tgcggcgtcg
acacctcgcc gccaccgccg 720ttcttccacg accactccat ctccatcaag caagcatact
acggatcaac cggcgcccac 780caccaccacc acgcgatcgc caccatggac ggatcaagct
taataggaga tcatcaccat 840cacagcagca gcatcctctt tggcggcgca tcagtgccac
ctctcctaga ccaccaaacc 900attctcgacg acgacgacga ccaccctaac aaaaccggca
gcaacacgac cgcggccaca 960ctgagcagca acatcacaga caacagcaac agcaacaaga
acaacagtga taataacaac 1020aacatcagca gcagctgctg cattagccta atgaacagca
gcagcaacat gatctattgg 1080gagggtcacc accaacaaca gcagcagcag catcagatgc
tgcagcagca gcagcagcac 1140atgagcagga atgtcatggg agagtgggac ttggaggagc
tgatgaaaga tgtgtcatcc 1200ttgcctttcc ttgatttcca agttgaa
122760409PRTOryza sativaLOC_Os12g33070.1 60Met Arg
Lys Pro Asp Cys Gly Gly Gly Gly Gly Ala Ala Lys Gly Gly 1 5
10 15 Gly Val Leu Gly Val Ala Gly
Gly Asn Asn Ala Ala Val Val Gly Gly 20 25
30 Lys Val Arg Lys Gly Leu Trp Ser Pro Glu Glu Asp
Glu Lys Leu Val 35 40 45
Ala Tyr Met Leu Arg Ser Gly Gln Gly Ser Trp Ser Asp Val Ala Arg
50 55 60 Asn Ala Gly
Leu Gln Arg Cys Gly Lys Ser Cys Arg Leu Arg Trp Ile 65
70 75 80 Asn Tyr Leu Arg Pro Asp Leu
Lys Arg Gly Ala Phe Ser Pro Gln Glu 85
90 95 Glu Asp Leu Ile Val Asn Leu His Ala Ile Leu
Gly Asn Arg Trp Ser 100 105
110 Gln Ile Ala Ala Arg Leu Pro Gly Arg Thr Asp Asn Glu Ile Lys
Asn 115 120 125 Phe
Trp Asn Ser Thr Ile Lys Lys Arg Leu Lys Ile Ser Ser Ser Ser 130
135 140 Ala Ser Pro Ala Thr Thr
Thr Asp Cys Ala Ser Pro Pro Glu His Lys 145 150
155 160 Leu Gly Ala Val Val Asp Leu Ala Gly Gly Gly
Gly Ala Thr Asp Asp 165 170
175 Val Val Val Gly Thr Ala Asn Ala Ala Met Lys Ser Met Trp Val Asp
180 185 190 Ser Ser
Ser Ser Ser Ser Ser Ser Ser Ser Ser Met Gln Ser Arg Pro 195
200 205 Ser Ile Met Ala Ala Ala Ala
Ala Gly Arg Ser Tyr Gly Gly Leu Leu 210 215
220 Pro Leu Pro Asp Gln Val Cys Gly Val Asp Thr Ser
Pro Pro Pro Pro 225 230 235
240 Phe Phe His Asp His Ser Ile Ser Ile Lys Gln Ala Tyr Tyr Gly Ser
245 250 255 Thr Gly Ala
His His His His His Ala Ile Ala Thr Met Asp Gly Ser 260
265 270 Ser Leu Ile Gly Asp His His His
His Ser Ser Ser Ile Leu Phe Gly 275 280
285 Gly Ala Ser Val Pro Pro Leu Leu Asp His Gln Thr Ile
Leu Asp Asp 290 295 300
Asp Asp Asp His Pro Asn Lys Thr Gly Ser Asn Thr Thr Ala Ala Thr 305
310 315 320 Leu Ser Ser Asn
Ile Thr Asp Asn Ser Asn Ser Asn Lys Asn Asn Ser 325
330 335 Asp Asn Asn Asn Asn Ile Ser Ser Ser
Cys Cys Ile Ser Leu Met Asn 340 345
350 Ser Ser Ser Asn Met Ile Tyr Trp Glu Gly His His Gln Gln
Gln Gln 355 360 365
Gln Gln His Gln Met Leu Gln Gln Gln Gln Gln His Met Ser Arg Asn 370
375 380 Val Met Gly Glu Trp
Asp Leu Glu Glu Leu Met Lys Asp Val Ser Ser 385 390
395 400 Leu Pro Phe Leu Asp Phe Gln Val Glu
405 6161PRTArabidopsis thalianaAT5G52260.1
1st Myb Domain 61Trp Ser Pro Glu Glu Asp Gln Lys Leu Lys Ser Phe Ile Leu
Ser Arg 1 5 10 15
Gly His Ala Cys Trp Thr Thr Val Pro Ile Leu Ala Gly Leu Gln Arg
20 25 30 Asn Gly Lys Ser Cys
Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro Gly 35
40 45 Leu Lys Arg Gly Ser Phe Ser Glu Glu
Glu Glu Glu Thr 50 55 60
6261PRTArabidopsis thalianaAT4G25560.1 1st Myb Domain 62Trp Ser Pro Glu
Glu Asp Glu Lys Leu Arg Ser Phe Ile Leu Ser Tyr 1 5
10 15 Gly His Ser Cys Trp Thr Thr Val Pro
Ile Lys Ala Gly Leu Gln Arg 20 25
30 Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg
Pro Gly 35 40 45
Leu Lys Arg Asp Met Ile Ser Ala Glu Glu Glu Glu Thr 50
55 60 6361PRTOryza sativaLOC_Os04g45020.1 1st Myb
Domain 63Trp Ser Pro Glu Glu Asp Gln Lys Leu Arg Asp Phe Ile Leu Arg Tyr
1 5 10 15 Gly His
Gly Cys Trp Ser Ala Val Pro Val Lys Ala Gly Leu Gln Arg 20
25 30 Asn Gly Lys Ser Cys Arg Leu
Arg Trp Ile Asn Tyr Leu Arg Pro Gly 35 40
45 Leu Lys His Gly Met Phe Ser Arg Glu Glu Glu Glu
Thr 50 55 60
6461PRTBrachypodium distachyonBradi5g16672.1 1st Myb Domain 64Trp Ser Pro
Glu Glu Asp Gln Lys Leu Arg Asp Tyr Ile Ile Arg Tyr 1 5
10 15 Gly His Ser Cys Trp Ser Thr Val
Pro Val Lys Ala Gly Leu Gln Arg 20 25
30 Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu
Arg Pro Gly 35 40 45
Leu Lys His Gly Met Phe Ser Gln Glu Glu Glu Glu Thr 50
55 60 6561PRTZea maysGRMZM2G170049_T01 1st Myb
Domain 65Trp Ser Pro Glu Glu Asp Gln Lys Leu Arg Asp Tyr Ile Leu Leu His
1 5 10 15 Gly His
Gly Cys Trp Ser Ala Leu Pro Ala Lys Ala Gly Leu Gln Arg 20
25 30 Asn Gly Lys Ser Cys Arg Leu
Arg Trp Ile Asn Tyr Leu Arg Pro Gly 35 40
45 Leu Lys His Gly Met Phe Ser Pro Glu Glu Glu Glu
Thr 50 55 60 6661PRTSetaria
italicaSi012304m 1st Myb Domain 66Trp Ser Pro Glu Glu Asp Glu Lys Leu Arg
Asp Phe Ile Leu Arg Tyr 1 5 10
15 Gly His Gly Cys Trp Ser Ala Leu Pro Ala Lys Ala Gly Leu Gln
Arg 20 25 30 Asn
Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro Gly 35
40 45 Leu Lys His Gly Met Phe
Ser Arg Glu Glu Glu Glu Thr 50 55
60 6761PRTCitrus clementinaclementine0.9_033485m 1st Myb Domain
67Trp Ser Pro Glu Glu Asp Gln Arg Leu Lys Asn Tyr Val Leu Gln His 1
5 10 15 Gly His Pro Cys
Trp Ser Ser Val Pro Ile Asn Ala Gly Leu Gln Arg 20
25 30 Asn Gly Lys Ser Cys Arg Leu Arg Trp
Ile Asn Tyr Leu Arg Pro Gly 35 40
45 Leu Lys Arg Gly Val Phe Asn Met Gln Glu Glu Glu Thr
50 55 60 6861PRTPopulus
trichocarpaPOPTR_0015s13190.1 1st Myb Domain 68Trp Ser Pro Glu Glu Asp
Gln Arg Leu Arg Asn Tyr Val Leu Lys His 1 5
10 15 Gly His Gly Cys Trp Ser Ser Val Pro Ile Asn
Ala Gly Leu Gln Arg 20 25
30 Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro
Gly 35 40 45 Leu
Lys Arg Gly Thr Phe Ser Ala Gln Glu Glu Glu Thr 50
55 60 6961PRTEucalyptus grandisEUCGR.K00250.1 1st
Myb Domain 69Trp Ser Pro Glu Glu Asp Gln Lys Leu Arg Asn Tyr Val Leu Lys
His 1 5 10 15 Gly
His Gly Cys Trp Ser Ser Val Pro Ile Asn Thr Gly Leu Gln Arg
20 25 30 Asn Gly Lys Ser Cys
Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro Gly 35
40 45 Leu Lys Arg Gly Met Phe Thr Met Glu
Glu Glu Glu Ile 50 55 60
7061PRTEucalyptus grandisEUCGR.K00251.1 1st Myb Domain 70Trp Ser Pro Glu
Glu Asp Gln Arg Leu Arg Asn Tyr Ile Leu Asn His 1 5
10 15 Gly His Gly Tyr Trp Ser Ser Val Pro
Ile Asn Thr Gly Leu Gln Arg 20 25
30 Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg
Pro Gly 35 40 45
Leu Lys Arg Gly Met Phe Thr Leu Glu Glu Glu Glu Ile 50
55 60 7161PRTPopulus
trichocarpaPOPTR_0012s13260.1 1st Myb Domain 71Trp Ser Pro Glu Glu Asp
Gln Arg Leu Gly Ser Tyr Val Phe Gln His 1 5
10 15 Gly His Gly Cys Trp Ser Ser Val Pro Ile Asn
Ala Gly Leu Gln Arg 20 25
30 Thr Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro
Gly 35 40 45 Leu
Lys Arg Gly Ala Phe Ser Thr Asp Glu Glu Glu Thr 50
55 60 7261PRTGlycine maxGlyma16g31280.1 1st Myb
Domain 72Trp Ser Pro Glu Glu Asp Asn Lys Leu Arg Asn His Ile Ile Lys His
1 5 10 15 Gly His
Gly Cys Trp Ser Ser Val Pro Ile Lys Ala Gly Leu Gln Arg 20
25 30 Asn Gly Lys Ser Cys Arg Leu
Arg Trp Ile Asn Tyr Leu Arg Pro Gly 35 40
45 Leu Lys Arg Gly Val Phe Ser Lys His Glu Glu Asp
Thr 50 55 60 7361PRTGlycine
maxGlyma09g25590.1 1st Myb Domain 73Trp Ser Pro Glu Glu Asp Asn Lys Leu
Arg Asn His Ile Ile Lys His 1 5 10
15 Gly His Gly Cys Trp Ser Ser Val Pro Ile Lys Ala Gly Leu
Gln Arg 20 25 30
Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro Gly
35 40 45 Leu Lys Arg Gly
Val Phe Ser Lys His Glu Lys Asp Thr 50 55
60 7461PRTSolanum lycopersicumSolyc03g025870.2.1 1st Myb
Domain 74Trp Ser Pro Asp Glu Asp Asp Arg Leu Lys Asn Tyr Met Ile Lys His
1 5 10 15 Gly His
Gly Cys Trp Ser Ser Val Pro Ile Asn Ala Gly Leu Gln Arg 20
25 30 Asn Gly Lys Ser Cys Arg Leu
Arg Trp Ile Asn Tyr Leu Arg Pro Gly 35 40
45 Leu Lys Arg Gly Ala Phe Ser Leu Glu Glu Glu Asp
Ile 50 55 60 7561PRTVitis
viniferaGSVIVT01028984001 1st Myb Domain 75Trp Ser Pro Glu Glu Asp Ala
Arg Leu Arg Asn Tyr Val Leu Lys Tyr 1 5
10 15 Gly Leu Gly Cys Trp Ser Ser Val Pro Val Asn
Ala Gly Leu Gln Arg 20 25
30 Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro
Gly 35 40 45 Leu
Lys Arg Gly Met Phe Thr Ile Glu Glu Glu Glu Thr 50
55 60 7661PRTEucalyptus grandisEUCGR.A02796.1 1st
Myb Domain 76Trp Ser Pro Asp Glu Asp Gln Arg Leu Arg Asn Tyr Ile His Lys
His 1 5 10 15 Gly
Tyr Ser Cys Trp Ser Ser Val Pro Ile Asn Ala Gly Leu Gln Arg
20 25 30 Asn Gly Lys Ser Cys
Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro Gly 35
40 45 Leu Lys Arg Gly Ala Phe Thr Val Gln
Glu Glu Glu Thr 50 55 60
7761PRTArabidopsis thalianaAT3G48920.1 1st Myb Domain 77Trp Ser Pro Glu
Glu Asp Glu Lys Leu Arg Ser His Val Leu Lys Tyr 1 5
10 15 Gly His Gly Cys Trp Ser Thr Ile Pro
Leu Gln Ala Gly Leu Gln Arg 20 25
30 Asn Gly Lys Ser Cys Arg Leu Arg Trp Val Asn Tyr Leu Arg
Pro Gly 35 40 45
Leu Lys Lys Ser Leu Phe Thr Lys Gln Glu Glu Thr Ile 50
55 60 7846PRTArabidopsis thalianaAT5G52260.1
SANT1 domai 78Leu Trp Ser Pro Glu Glu Asp Gln Lys Leu Lys Ser Phe Ile Leu
Ser 1 5 10 15 Arg
Gly His Ala Cys Trp Thr Thr Val Pro Ile Leu Ala Gly Leu Gln
20 25 30 Arg Asn Gly Lys Ser
Cys Arg Leu Arg Trp Ile Asn Tyr Leu 35 40
45 7946PRTArabidopsis thalianaAT4G25560.1 SANT1 domain
79Leu Trp Ser Pro Glu Glu Asp Glu Lys Leu Arg Ser Phe Ile Leu Ser 1
5 10 15 Tyr Gly His Ser
Cys Trp Thr Thr Val Pro Ile Lys Ala Gly Leu Gln 20
25 30 Arg Asn Gly Lys Ser Cys Arg Leu Arg
Trp Ile Asn Tyr Leu 35 40 45
8046PRTOryza sativaLOC_Os04g45020.1 SANT1 domain 80Leu Trp Ser Pro Glu
Glu Asp Gln Lys Leu Arg Asp Phe Ile Leu Arg 1 5
10 15 Tyr Gly His Gly Cys Trp Ser Ala Val Pro
Val Lys Ala Gly Leu Gln 20 25
30 Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu
35 40 45 8146PRTBrachypodium
distachyonBradi5g16672.1 SANT1 domain 81Leu Trp Ser Pro Glu Glu Asp Gln
Lys Leu Arg Asp Tyr Ile Ile Arg 1 5 10
15 Tyr Gly His Ser Cys Trp Ser Thr Val Pro Val Lys Ala
Gly Leu Gln 20 25 30
Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu 35
40 45 8246PRTZea maysGRMZM2G170049_T01
SANT1 domain 82Leu Trp Ser Pro Glu Glu Asp Gln Lys Leu Arg Asp Tyr Ile
Leu Leu 1 5 10 15
His Gly His Gly Cys Trp Ser Ala Leu Pro Ala Lys Ala Gly Leu Gln
20 25 30 Arg Asn Gly Lys Ser
Cys Arg Leu Arg Trp Ile Asn Tyr Leu 35 40
45 8346PRTSetaria italicaSi012304m SANT1 domain 83Leu Trp
Ser Pro Glu Glu Asp Glu Lys Leu Arg Asp Phe Ile Leu Arg 1 5
10 15 Tyr Gly His Gly Cys Trp Ser
Ala Leu Pro Ala Lys Ala Gly Leu Gln 20 25
30 Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn
Tyr Leu 35 40 45
8446PRTCitrus clementinaclementine0.9_033485m SANT1 domain 84Leu Trp Ser
Pro Glu Glu Asp Gln Arg Leu Lys Asn Tyr Val Leu Gln 1 5
10 15 His Gly His Pro Cys Trp Ser Ser
Val Pro Ile Asn Ala Gly Leu Gln 20 25
30 Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr
Leu 35 40 45
8546PRTPopulus trichocarpaPOPTR_0015s13190.1 SANT1 domain 85Leu Trp Ser
Pro Glu Glu Asp Gln Arg Leu Arg Asn Tyr Val Leu Lys 1 5
10 15 His Gly His Gly Cys Trp Ser Ser
Val Pro Ile Asn Ala Gly Leu Gln 20 25
30 Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr
Leu 35 40 45
8646PRTEucalyptus grandisEucgr.K00250.1 SANT1 domain 86Leu Trp Ser Pro
Glu Glu Asp Gln Lys Leu Arg Asn Tyr Val Leu Lys 1 5
10 15 His Gly His Gly Cys Trp Ser Ser Val
Pro Ile Asn Thr Gly Leu Gln 20 25
30 Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu
35 40 45 8746PRTEucalyptus
grandisEucgr.K00251.1 SANT1 domain 87Leu Trp Ser Pro Glu Glu Asp Gln Arg
Leu Arg Asn Tyr Ile Leu Asn 1 5 10
15 His Gly His Gly Tyr Trp Ser Ser Val Pro Ile Asn Thr Gly
Leu Gln 20 25 30
Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu 35
40 45 8846PRTPopulus
trichocarpaPOPTR_0012s13260.1 SANT1 domain 88Leu Trp Ser Pro Glu Glu Asp
Gln Arg Leu Gly Ser Tyr Val Phe Gln 1 5
10 15 His Gly His Gly Cys Trp Ser Ser Val Pro Ile
Asn Ala Gly Leu Gln 20 25
30 Arg Thr Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu
35 40 45 8946PRTGlycine
maxGlyma16g31280.1 SANT1 domain 89Leu Trp Ser Pro Glu Glu Asp Asn Lys Leu
Arg Asn His Ile Ile Lys 1 5 10
15 His Gly His Gly Cys Trp Ser Ser Val Pro Ile Lys Ala Gly Leu
Gln 20 25 30 Arg
Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu 35
40 45 9046PRTGlycine maxGlyma09g25590.1 SANT1
domain 90Leu Trp Ser Pro Glu Glu Asp Asn Lys Leu Arg Asn His Ile Ile Lys
1 5 10 15 His Gly
His Gly Cys Trp Ser Ser Val Pro Ile Lys Ala Gly Leu Gln 20
25 30 Arg Asn Gly Lys Ser Cys Arg
Leu Arg Trp Ile Asn Tyr Leu 35 40
45 9146PRTSolanum lycopersicumSolyc03g025870.2.1 SANT1 domain
91Leu Trp Ser Pro Asp Glu Asp Asp Arg Leu Lys Asn Tyr Met Ile Lys 1
5 10 15 His Gly His Gly
Cys Trp Ser Ser Val Pro Ile Asn Ala Gly Leu Gln 20
25 30 Arg Asn Gly Lys Ser Cys Arg Leu Arg
Trp Ile Asn Tyr Leu 35 40 45
9246PRTVitis viniferaGSVIVT01028984001 SANT1 domain 92Leu Trp Ser Pro
Glu Glu Asp Ala Arg Leu Arg Asn Tyr Val Leu Lys 1 5
10 15 Tyr Gly Leu Gly Cys Trp Ser Ser Val
Pro Val Asn Ala Gly Leu Gln 20 25
30 Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu
35 40 45 9346PRTEucalyptus
grandisEucgr.A02796.1 SANT1 domain 93Leu Trp Ser Pro Asp Glu Asp Gln Arg
Leu Arg Asn Tyr Ile His Lys 1 5 10
15 His Gly Tyr Ser Cys Trp Ser Ser Val Pro Ile Asn Ala Gly
Leu Gln 20 25 30
Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu 35
40 45 9446PRTArabidopsis
thalianaAT3G48920.1 SANT1 domain 94Leu Trp Ser Pro Glu Glu Asp Glu Lys
Leu Arg Ser His Val Leu Lys 1 5 10
15 Tyr Gly His Gly Cys Trp Ser Thr Ile Pro Leu Gln Ala Gly
Leu Gln 20 25 30
Arg Asn Gly Lys Ser Cys Arg Leu Arg Trp Val Asn Tyr Leu 35
40 45 9543PRTArabidopsis
thalianaAT5G52260.1 2nd Myb Domain 95Phe Ser Glu Glu Glu Glu Glu Thr Ile
Leu Thr Leu His Ser Ser Leu 1 5 10
15 Gly Asn Lys Trp Ser Arg Ile Ala Lys Tyr Leu Pro Gly Arg
Thr Asp 20 25 30
Asn Glu Ile Lys Asn Tyr Trp His Ser Tyr Leu 35
40 9643PRTArabidopsis thalianaAT4G25560.1 2nd Myb Domain
96Ile Ser Ala Glu Glu Glu Glu Thr Ile Leu Thr Phe His Ser Ser Leu 1
5 10 15 Gly Asn Lys Trp
Ser Gln Ile Ala Lys Phe Leu Pro Gly Arg Thr Asp 20
25 30 Asn Glu Ile Lys Asn Tyr Trp His Ser
His Leu 35 40 9743PRTOryza
sativaLOC_Os04g45020.1 2nd Myb Domain 97Phe Ser Arg Glu Glu Glu Glu Thr
Val Met Asn Leu His Ala Thr Met 1 5 10
15 Gly Asn Lys Trp Ser Gln Ile Ala Arg His Leu Pro Gly
Arg Thr Asp 20 25 30
Asn Glu Val Lys Asn Tyr Trp Asn Ser Tyr Leu 35
40 9843PRTBrachypodium distachyonBradi5g16672.1 2nd Myb
Domain 98Phe Ser Gln Glu Glu Glu Glu Thr Val Met Ser Leu His Ala Thr Leu
1 5 10 15 Gly Asn
Lys Trp Ser Arg Ile Ala Gln His Leu Pro Gly Arg Thr Asp 20
25 30 Asn Glu Val Lys Asn Tyr Trp
Asn Ser Tyr Leu 35 40 9943PRTZea
maysGRMZM2G170049_T01 2nd Myb Domain 99Phe Ser Pro Glu Glu Glu Glu Thr
Val Met Ser Leu His Ala Thr Leu 1 5 10
15 Gly Asn Lys Trp Ser Arg Ile Ala Arg His Leu Pro Gly
Arg Thr Asp 20 25 30
Asn Glu Val Lys Asn Tyr Trp Asn Ser Tyr Leu 35
40 10043PRTSetaria italicaSi012304m 2nd Myb Domain 100Phe
Ser Arg Glu Glu Glu Glu Thr Val Met Ser Leu His Ala Lys Leu 1
5 10 15 Gly Asn Lys Trp Ser Gln
Ile Ala Arg His Leu Pro Gly Arg Thr Asp 20
25 30 Asn Glu Val Lys Asn Tyr Trp Asn Ser Tyr
Leu 35 40 10143PRTCitrus
clementinaclementine0.9_033485m 2nd Myb Domain 101Phe Asn Met Gln Glu Glu
Glu Thr Ile Leu Thr Val His Arg Leu Leu 1 5
10 15 Gly Asn Lys Trp Ser Gln Ile Ala Gln His Leu
Pro Gly Arg Thr Asp 20 25
30 Asn Glu Ile Lys Asn Tyr Trp His Ser His Leu 35
40 10243PRTPopulus trichocarpaPOPTR_0015s13190.1
2nd Myb Domain 102Phe Ser Ala Gln Glu Glu Glu Thr Ile Leu Ala Leu His His
Met Leu 1 5 10 15
Gly Asn Lys Trp Ser Gln Ile Ala Gln His Leu Pro Gly Arg Thr Asp
20 25 30 Asn Glu Ile Lys Asn
His Trp His Ser Tyr Leu 35 40
10343PRTEucalyptus grandisEUCGR.K00250.1 2nd Myb Domain 103Phe Thr Met
Glu Glu Glu Glu Ile Ile Phe Ser Leu His His Leu Ile 1 5
10 15 Gly Asn Lys Trp Ser Gln Ile Ala
Lys His Leu Pro Gly Arg Thr Asp 20 25
30 Asn Glu Ile Lys Asn His Trp His Ser Tyr Leu
35 40 10443PRTEucalyptus
grandisEUCGR.K00251.1 2nd Myb Domain 104Phe Thr Leu Glu Glu Glu Glu Ile
Ile Leu Ser Leu His Arg Leu Ile 1 5 10
15 Gly Asn Lys Trp Ser Gln Ile Ala Lys His Leu Pro Gly
Arg Thr Asp 20 25 30
Asn Glu Ile Lys Asn His Trp His Ser Tyr Leu 35
40 10543PRTPopulus trichocarpaPOPTR_0012s13260.1 2nd Myb
Domain 105Phe Ser Thr Asp Glu Glu Glu Thr Ile Leu Thr Leu His Arg Met Leu
1 5 10 15 Gly Asn
Lys Trp Ser Gln Ile Ala Gln His Leu Pro Gly Arg Thr Asp 20
25 30 Asn Glu Ile Lys Asn His Trp
His Ser Tyr Leu 35 40
10643PRTGlycine maxGlyma16g31280.1 2nd Myb Domain 106Phe Ser Lys His Glu
Glu Asp Thr Ile Met Val Leu His His Met Leu 1 5
10 15 Gly Asn Lys Trp Ser Gln Ile Ala Gln His
Leu Pro Gly Arg Thr Asp 20 25
30 Asn Glu Ile Lys Asn Tyr Trp His Ser Tyr Leu 35
40 10743PRTGlycine maxGlyma09g25590.1 2nd Myb
Domain 107Phe Ser Lys His Glu Lys Asp Thr Ile Met Ala Leu His His Met Leu
1 5 10 15 Gly Asn
Lys Trp Ser Gln Ile Ala Gln His Leu Pro Gly Arg Thr Asp 20
25 30 Asn Glu Val Lys Asn Tyr Trp
His Ser Tyr Leu 35 40
10843PRTSolanum lycopersicumSolyc03g025870.2.1 2nd Myb Domain 108Phe Ser
Leu Glu Glu Glu Asp Ile Ile Leu Thr Leu His Ala Met Phe 1 5
10 15 Gly Asn Lys Trp Ser Gln Ile
Ala Gln Gln Leu Pro Gly Arg Thr Asp 20 25
30 Asn Glu Ile Lys Asn His Trp His Ser Tyr Leu
35 40 10943PRTVitis
viniferaGSVIVT01028984001 2nd Myb Domain 109Phe Thr Ile Glu Glu Glu Glu
Thr Ile Met Ala Leu His Arg Leu Leu 1 5
10 15 Gly Asn Lys Trp Ser Gln Ile Ala Gln Asn Phe
Pro Gly Arg Thr Asp 20 25
30 Asn Glu Ile Lys Asn Tyr Trp His Ser Cys Leu 35
40 11043PRTEucalyptus grandisEUCGR.A02796.1 2nd
Myb Domain 110Phe Thr Val Gln Glu Glu Glu Thr Ile Leu Asn Leu His His Leu
Leu 1 5 10 15 Gly
Asn Lys Trp Ser Gln Ile Ala Gln His Leu Pro Gly Arg Thr Asp
20 25 30 Asn Glu Ile Lys Asn
His Trp His Ser Tyr Leu 35 40
11143PRTArabidopsis thalianaAT3G48920.1 2nd Myb Domain 111Phe Thr Lys Gln
Glu Glu Thr Ile Leu Leu Ser Leu His Ser Met Leu 1 5
10 15 Gly Asn Lys Trp Ser Gln Ile Ser Lys
Phe Leu Pro Gly Arg Thr Asp 20 25
30 Asn Glu Ile Lys Asn Tyr Trp His Ser Asn Leu 35
40 11244PRTArabidopsis thalianaAT4G25560.1
SANT2 domain 112Met Ile Ser Ala Glu Glu Glu Glu Thr Ile Leu Thr Phe His
Ser Ser 1 5 10 15
Leu Gly Asn Lys Trp Ser Gln Ile Ala Lys Phe Leu Pro Gly Arg Thr
20 25 30 Asp Asn Glu Ile Lys
Asn Tyr Trp His Ser His Leu 35 40
11344PRTArabidopsis thalianaAT5G52260.1 SANT2 domain 113Ser Phe Ser Glu
Glu Glu Glu Glu Thr Ile Leu Thr Leu His Ser Ser 1 5
10 15 Leu Gly Asn Lys Trp Ser Arg Ile Ala
Lys Tyr Leu Pro Gly Arg Thr 20 25
30 Asp Asn Glu Ile Lys Asn Tyr Trp His Ser Tyr Leu
35 40 11444PRTOryza
sativaLOC_Os04g45020.1 SANT2 domain 114Met Phe Ser Arg Glu Glu Glu Glu
Thr Val Met Asn Leu His Ala Thr 1 5 10
15 Met Gly Asn Lys Trp Ser Gln Ile Ala Arg His Leu Pro
Gly Arg Thr 20 25 30
Asp Asn Glu Val Lys Asn Tyr Trp Asn Ser Tyr Leu 35
40 11544PRTBrachypodium distachyonBradi5g16672.1
SANT2 domain 115Met Phe Ser Gln Glu Glu Glu Glu Thr Val Met Ser Leu His
Ala Thr 1 5 10 15
Leu Gly Asn Lys Trp Ser Arg Ile Ala Gln His Leu Pro Gly Arg Thr
20 25 30 Asp Asn Glu Val Lys
Asn Tyr Trp Asn Ser Tyr Leu 35 40
11644PRTZea maysGRMZM2G170049_T01 SANT2 domain 116Met Phe Ser Pro Glu
Glu Glu Glu Thr Val Met Ser Leu His Ala Thr 1 5
10 15 Leu Gly Asn Lys Trp Ser Arg Ile Ala Arg
His Leu Pro Gly Arg Thr 20 25
30 Asp Asn Glu Val Lys Asn Tyr Trp Asn Ser Tyr Leu 35
40 11744PRTSetaria italicaSi012304m
SANT2 domain 117Met Phe Ser Arg Glu Glu Glu Glu Thr Val Met Ser Leu His
Ala Lys 1 5 10 15
Leu Gly Asn Lys Trp Ser Gln Ile Ala Arg His Leu Pro Gly Arg Thr
20 25 30 Asp Asn Glu Val Lys
Asn Tyr Trp Asn Ser Tyr Leu 35 40
11844PRTCitrus clementinaclementine0.9_033485m SANT2 domain 118Val Phe
Asn Met Gln Glu Glu Glu Thr Ile Leu Thr Val His Arg Leu 1 5
10 15 Leu Gly Asn Lys Trp Ser Gln
Ile Ala Gln His Leu Pro Gly Arg Thr 20 25
30 Asp Asn Glu Ile Lys Asn Tyr Trp His Ser His Leu
35 40 11944PRTPopulus
trichocarpaPOPTR_0015s13190.1 SANT2 domain 119Thr Phe Ser Ala Gln Glu Glu
Glu Thr Ile Leu Ala Leu His His Met 1 5
10 15 Leu Gly Asn Lys Trp Ser Gln Ile Ala Gln His
Leu Pro Gly Arg Thr 20 25
30 Asp Asn Glu Ile Lys Asn His Trp His Ser Tyr Leu 35
40 12044PRTEucalyptus
grandisEucgr.K00250.1 SANT2 domain 120Met Phe Thr Met Glu Glu Glu Glu Ile
Ile Phe Ser Leu His His Leu 1 5 10
15 Ile Gly Asn Lys Trp Ser Gln Ile Ala Lys His Leu Pro Gly
Arg Thr 20 25 30
Asp Asn Glu Ile Lys Asn His Trp His Ser Tyr Leu 35
40 12144PRTEucalyptus grandisEucgr.K00251.1 SANT2
domain 121Met Phe Thr Leu Glu Glu Glu Glu Ile Ile Leu Ser Leu His Arg Leu
1 5 10 15 Ile Gly
Asn Lys Trp Ser Gln Ile Ala Lys His Leu Pro Gly Arg Thr 20
25 30 Asp Asn Glu Ile Lys Asn His
Trp His Ser Tyr Leu 35 40
12244PRTPopulus trichocarpaPOPTR_0012s13260.1 SANT2 domain 122Ala Phe Ser
Thr Asp Glu Glu Glu Thr Ile Leu Thr Leu His Arg Met 1 5
10 15 Leu Gly Asn Lys Trp Ser Gln Ile
Ala Gln His Leu Pro Gly Arg Thr 20 25
30 Asp Asn Glu Ile Lys Asn His Trp His Ser Tyr Leu
35 40 12344PRTGlycine
maxGlyma16g31280.1 SANT2 domain 123Val Phe Ser Lys His Glu Glu Asp Thr
Ile Met Val Leu His His Met 1 5 10
15 Leu Gly Asn Lys Trp Ser Gln Ile Ala Gln His Leu Pro Gly
Arg Thr 20 25 30
Asp Asn Glu Ile Lys Asn Tyr Trp His Ser Tyr Leu 35
40 12444PRTGlycine maxGlyma09g25590.1 SANT2 domain
124Val Phe Ser Lys His Glu Lys Asp Thr Ile Met Ala Leu His His Met 1
5 10 15 Leu Gly Asn Lys
Trp Ser Gln Ile Ala Gln His Leu Pro Gly Arg Thr 20
25 30 Asp Asn Glu Val Lys Asn Tyr Trp His
Ser Tyr Leu 35 40
12544PRTSolanum lycopersicumSolyc03g025870.2.1 SANT2 domain 125Ala Phe
Ser Leu Glu Glu Glu Asp Ile Ile Leu Thr Leu His Ala Met 1 5
10 15 Phe Gly Asn Lys Trp Ser Gln
Ile Ala Gln Gln Leu Pro Gly Arg Thr 20 25
30 Asp Asn Glu Ile Lys Asn His Trp His Ser Tyr Leu
35 40 12644PRTVitis
viniferaGSVIVT01028984001 SANT2 domain 126Met Phe Thr Ile Glu Glu Glu Glu
Thr Ile Met Ala Leu His Arg Leu 1 5 10
15 Leu Gly Asn Lys Trp Ser Gln Ile Ala Gln Asn Phe Pro
Gly Arg Thr 20 25 30
Asp Asn Glu Ile Lys Asn Tyr Trp His Ser Cys Leu 35
40 12744PRTEucalyptus grandisEucgr.A02796.1 SANT2
domain 127Ala Phe Thr Val Gln Glu Glu Glu Thr Ile Leu Asn Leu His His Leu
1 5 10 15 Leu Gly
Asn Lys Trp Ser Gln Ile Ala Gln His Leu Pro Gly Arg Thr 20
25 30 Asp Asn Glu Ile Lys Asn His
Trp His Ser Tyr Leu 35 40
12844PRTArabidopsis thalianaAT3G48920.1 SANT2 domain 128Leu Phe Thr Lys
Gln Glu Glu Thr Ile Leu Leu Ser Leu His Ser Met 1 5
10 15 Leu Gly Asn Lys Trp Ser Gln Ile Ser
Lys Phe Leu Pro Gly Arg Thr 20 25
30 Asp Asn Glu Ile Lys Asn Tyr Trp His Ser Asn Leu
35 40 12958PRTArabidopsis
thalianamisc_feature(4)..(4)Xaa is Asp or Glu 129Trp Ser Pro Xaa Glu Asp
Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5
10 15 Gly Xaa Xaa Xaa Trp Xaa Xaa Xaa Pro Xaa Xaa
Xaa Gly Leu Gln Arg 20 25
30 Xaa Gly Lys Ser Cys Arg Leu Arg Trp Xaa Asn Tyr Leu Arg Pro
Gly 35 40 45 Leu
Lys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Glu 50 55
13039PRTArabidopsis thalianamisc_feature(2)..(4)Xaa can be any
naturally occurring amino acid 130Glu Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Xaa
Xaa Xaa Gly Asn Lys Trp 1 5 10
15 Ser Xaa Ile Xaa Xaa Xaa Xaa Pro Gly Arg Thr Asp Asn Glu Xaa
Lys 20 25 30 Asn
Xaa Trp Xaa Ser Xaa Leu 35 13146PRTArabidopsis
thalianamisc_feature(5)..(5)Xaa can be any naturally occurring amino acid
131Leu Trp Ser Pro Xaa Glu Asp Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa 1
5 10 15 Xaa Gly Xaa Xaa
Xaa Trp Xaa Xaa Xaa Pro Xaa Xaa Xaa Gly Leu Gln 20
25 30 Arg Xaa Gly Lys Ser Cys Arg Leu Arg
Trp Xaa Asn Tyr Leu 35 40 45
13239PRTArabidopsis thalianamisc_feature(2)..(8)Xaa can be any
naturally occurring amino acid 132Glu Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Xaa
Xaa Xaa Gly Asn Lys Trp 1 5 10
15 Ser Xaa Ile Xaa Xaa Xaa Xaa Pro Gly Arg Thr Asp Asn Glu Xaa
Lys 20 25 30 Asn
Xaa Trp Xaa Ser Xaa Leu 35 1331009DNASolanum
lycopersicumRBCS3 (Ribulose 1,5-bisphosphate carboxylase, small
subunit 3) leaf-specific promoter 133aaatggagta atatggataa tcaacgcaac
tatatagaga aaaaataata gcgctaccat 60atacgaaaaa tagtaaaaaa ttataataat
gattcagaat aaattattaa taactaaaaa 120gcgtaaagaa ataaattaga gaataagtga
tacaaaattg gatgttaatg gatacttctt 180ataattgctt aaaaggaata caagatggga
aataatgtgt tattattatt gatgtataaa 240gaatttgtac aatttttgta tcaataaagt
tccaaaaata atctttaaaa aataaaagta 300cccttttatg aactttttat caaataaatg
aaatccaata ttagcaaaac attgatatta 360ttactaaata tttgttaaat taaaaaatat
gtcattttat tttttaacag atatttttta 420aagtaaatgt tataaattac gaaaaaggga
ttaatgagta tcaaaacagc ctaaatggga 480ggagacaata acagaaattt gctgtagtaa
ggtggcttaa gtcatcattt aatttgatat 540tataaaaatt ctaattagtt tatagtcttt
cttttcctct tttgtttgtc ttgtatgcta 600aaaaaggtat attatatcta taaattatgt
agcataatga ccacatctgg catcatcttt 660acacaattca cctaaatatc tcaagcgaag
ttttgccaaa actgaagaaa agatttgaac 720aacctatcaa gtaacaaaaa tcccaaacaa
tatagtcatc tatattaaat cttttcaatt 780gaagaaattg tcaaagacac atacctctat
gagttttttc atcaattttt ttttcttttt 840taaactgtat ttttaaaaaa atattgaata
aaacatgtcc tattcattag tttgggaact 900ttaagataag gagtgtgtaa tttcagaggc
tattaatttt gaaatgtcaa gagccacata 960atccaatggt tatggttgct cttagatgag
gttattgctt taggtgaaa 10091341714DNAArabidopsis
thalianaRBCS4 leaf-specific promoter sequence 134caaatttatt atgtgttttt
tttccgtggt cgagattgtg tattattctt tagttattac 60aagactttta gctaaaattt
gaaagaattt actttaagaa aatcttaaca tctgagataa 120tttcagcaat agattatatt
tttcattact ctagcagtat ttttgcagat caatcgcaac 180atatatggtt gttagaaaaa
atgcactata tatatatata ttattttttc aattaaaagt 240gcatgatata taatatatat
atatatatat atgtgtgtgt gtatatggtc aaagaaattc 300ttatacaaat atacacgaac
acatatattt gacaaaatca aagtattaca ctaaacaatg 360agttggtgca tggccaaaac
aaatatgtag attaaaaatt ccagcctcca aaaaaaaatc 420caagtgttgt aaagcattat
atatatatag tagatcccaa atttttgtac aattccacac 480tgatcgaatt tttaaagttg
aatatctgac gtaggatttt tttaatgtct tacctgacca 540tttactaata acattcatac
gttttcattt gaaatatcct ctataattat attgaatttg 600gcacataata agaaacctaa
ttggtgattt attttactag taaatttctg gtgatgggct 660ttctactaga aagctctcgg
aaaatcttgg accaaatcca tattccatga cttcgattgt 720taaccctatt agttttcaca
aacatactat caatatcatt gcaacggaaa aggtacaagt 780aaaacattca atccgatagg
gaagtgatgt aggaggttgg gaagacaggc ccagaaagag 840atttatctga cttgttttgt
gtatagtttt caatgttcat aaaggaagat ggagacttga 900gaagtttttt ttggactttg
tttagctttg ttgggcgttt ttttttttga tcaataactt 960tgttgggctt atgatttgta
atattttcgt ggactcttta gtttatttag acgtgctaac 1020tttgttgggc ttatgacttg
ttgtaacata ttgtaacaga tgacttgatg tgcgactaat 1080ctttacacat taaacatagt
tctgtttttt gaaagttctt attttcattt ttatttgaat 1140gttatatatt tttctatatt
tataattcta gtaaaaggca aattttgctt ttaaatgaaa 1200aaaatatata ttccacagtt
tcacctaatc ttatgcattt agcagtacaa attcaaaaat 1260ttcccatttt tattcatgaa
tcataccatt atatattaac taaatccaag gtaaaaaaaa 1320ggtatgaaag ctctatagta
agtaaaatat aaattcccca taaggaaagg gccaagtcca 1380ccaggcaagt aaaatgagca
agcaccactc caccatcaca caatttcact catagataac 1440gataagattc atggaattat
cttccacgtg gcattattcc agcggttcaa gccgataagg 1500gtctcaacac ctctccttag
gcctttgtgg ccgttaccaa gtaaaattaa cctcacacat 1560atccacactc aaaatccaac
ggtgtagatc ctagtccact tgaatctcat gtatcctaga 1620ccctccgatc actccaaagc
ttgttctcat tgttgttatc attatatata gatgaccaaa 1680gcactagacc aaacctcagt
cacacaaaga gtaa 17141351923DNAArabidopsis
thalianaAt4g01060 (G682) promoter sequence 135ttattaagtg ctatgcgtta
atcggcatct ataaagtgtt gcattgatga acaaagtgga 60tgcctaaact agacgtttaa
ctaaatgttt agaatgaaat cttcatctca tctaaaaagt 120gttgcattga tgtaaaaagt
ggatgcccat tagttcttgg ctttgaaatg tttttagaat 180gaaatcttca tcaatctcca
tatgtggttc aatccactca ttttatcttt tgttaaagat 240gttcttcagg ccaatataat
gatgaccatg gatggtttgc aactcgcata taacacttct 300ttatccgatg gttacaagta
ttacatggct atagatagct tttgcatgca acaaattatc 360tatcaaagtt tatgcatcct
ctaaaatatg gtcattggca agccactaaa cgtatatatt 420gtgacaatgt atgatgatat
atttttatgt gttgactccg tttttcatta agtaatgaaa 480catgttgctc tagattacca
ttttaatcgc aaacatatat gttgctctac cacgcattgc 540atcaaatgat aagcttgagg
acgcttcgac aaaaacatat ttctccttct tcttactgaa 600ccaaatgaca attgacaaac
cccattaata aaatcggtta gtgttaatgt gtcactcata 660atattaactt agtaaagaac
aagaccacat taattaaatc aggtgttagt ctgagaatat 720acgtttctct tcctcattcc
aaacttaaat tcggaattta ctgagaatat attgttagca 780ctgaaaaagg ttaagttgaa
agtttgctag ggatggcaat taaatatagt cttgccttgg 840ggatattccc ttcgtggact
tgtaagttta tttataggtc ctcttatgta tatatagatg 900atctaacgat cgatatacta
tgaaaaaagt tgttactaga ttttattgca ggtaatagtg 960ttgaataacc cgaaccaata
aagcagttgt aacgaacaca cgacacgttg cttactgcga 1020ggaccacttt gttttttgtt
ttttttggct ttaagccaat ttaggaccaa atttgcatga 1080ttgaggatgc aagtatccaa
cccattttca tctttcgtag tgacactcat ttacttttgt 1140gatggacacg ttatagtata
tcttaaatat taaagagaca tgattggggg atcattgttt 1200taatttaaat aatgtagatt
ctattctttt catggtatta atccaattta tagaaagtta 1260tgtgttatta gcaattaagc
taaatgatga aaacaatcag tttagtgaaa caaactcgcc 1320gagaaaacat gaatggttga
aaatattatt gtgttttaca aacgtacacg aggacaatag 1380ttttgtaagt ttttcttagg
cattgaaaaa tgtttgatac aaaaagtaat gttaaaataa 1440ttaaaaatga ttttgtctta
atatatccaa aatttcaatc tattatgaac aaagggagta 1500taatttctga ttgaatgaac
tggaatagca atcagaaaag ctttgaaaac aattgttgtt 1560gattattaat gatcttaatt
aacggcatgt atcaatattt atacaactta tgttccagtc 1620caagccatca caacggagta
aatgaagtca cgggtacttg tggtttttat tggttgcaaa 1680cttgcaactt gcaaagatag
ctaacaataa ttaatataat taatgagaac aaaaccaatt 1740tagtaaatta aaatccttta
acatagaaac cgaccaaacc cgttggaccg ttggttactt 1800gatttggtta gttgctataa
atagaaatga tggttcgtgt gcaaccttca aaatacgacc 1860actctctcag agtactctct
tagtttcttt cttcttcttc tttgtaatac ggtgccgttt 1920gac
19231361500DNAOryza
sativaOs02g09720 location Chr24997678-4999177 136ggccgtcgtc ggcgagttct
cagctatagc ttggtagcta tctagctcaa tttgctgtct 60ccaagtgtga cagctagttg
taattgagtt ggtatagctt tggcctcttc ttttattttt 120ttcaggccgt ttagatggta
acctctctga cttgagagag ttagagaggc cttgtatata 180cggagtatat agtactccag
tttagttgcc atgccagcag ttgcatgcac tagtaagcaa 240ctagtctctc ggtttcatat
tacaactcgt tttaactaag tttatagaaa aaacatagtc 300gtatttttaa tacaaaacaa
atatattatc aaaatatatt taatgtttgg tttaattaat 360taacattggt gtttttgatg
ttactaattt tttttataaa cttagttgaa cttaaaataa 420attggttaag aaaaagttaa
agcgacttgt aatatagaac ggaggaggta gctattggtc 480aaggggatgt gcatccatgt
ttggtaggta gccgtggtgg ctttggtgtg tcattctcta 540tcttttccac acacacactt
taaatttgct ggctttttag aagaacaatg gtaatgtttc 600gcttttattt tgctcatggt
aaaaagattg tgtttcgctt ttcgaggggc aaaaattaag 660tctaattgat gctattgatt
aataagatcg tgatcgcgag gaggattgag atgtctcgat 720ggacataata gaatcttagg
ttcaaggaat ttacactctt taataaagac ttcgcttgca 780aagatgtagg ccaaatgaaa
gagacttaag catgcttgtt gatataatac tagtttattt 840tgagaattag gatttaattt
gatgcactaa acctagagta aaaaccataa gatgacctaa 900atctgtgcca atatatttct
agttggttca taattaatca acggaatgag tatgcagttt 960ttctcttaaa atgcctatag
gagttaggtg ccatgctaca ccacactact gaaactgaat 1020tggtgccggt tggaaaccgg
ctatagatgt cggatatcct aactgatacc cagtagacga 1080cacctttggc ttcagtcatc
gatcccgccg gcatctttta cttacaaagg tgtcggtttt 1140gaatacagaa ccaacaccta
tactgtcata taggtgtcag ttcttaaatt gattggcgta 1200tggacagttt tacatgtgac
ttttacaggt gacagttcgt aaatacaact ggcacctata 1260agaagcatat atgtgccatt
tgtacaccgg gcaggacacg ggacaagggt tataggtgtt 1320tgtttgtaag taaaaccagc
acctatatgt tagcaagaaa aaaaataaaa atgataggac 1380ctaccaattt cacatacata
tcacacaata acagaaattc acatcacttc aaacatacaa 1440ccacattcac attcaaccaa
actatatata ttcaattcca catccacaac cctcatccca 15001371500DNAOryza
sativaOs05g34510 location Chr520455817-20457316 (reverse complement)
137ctatatcttc ctacctatcc ccctgcaagc taacaagggg aattacttgc acatctaatg
60atggactggt gttgttatgc ggttgtgttg gggtgagatg gaggatgtct ctcctccccc
120ttgctagcct ttaaaaagga gtcctacagg ggcataacag gaatgagagg aagcttctgg
180agatcaattc tcacaacaca catttcctac agtacagttt tgcccccaga cagatcggat
240ggtgtgtgtc agcagcctct atgcctgtag gcatgcattc agttgaatgg ttttgtctcg
300atcgacggcg aaaaaggatt tacttttgga caagaaaaaa gagcctttaa tccatgagat
360tactacatct cgcatacagg gacaagtatt acagctgtga gtgatgctta agagacttta
420cggttttgtt tgcagctaag caggaattaa tataaataat ctcccttttc agggctctgc
480ttgcatgaat agctaccagc cagagccaag agtgccagag acatcaagcc tggatggtag
540tagtacagta cttgcgttac tcgtgcccgg cgttgctgct tcggtggggc ccacttgaac
600tggccgtcct cccgagccgt cggtgggggt ggtccccacc accggccagt tactttgctc
660ccgtcgattc ggcccggtgc accaacagcc cattttttta tgatgggctt gcaacttggc
720ccaacttgtt ggattgtcaa ggccggaggg aggcccatct gggcgaactt gtgtgggccg
780cttctttcca ttttagagag gaaacatggg ctgggctacg ggctgcgggc tgccacctat
840caccggagaa acacttgctc gcagcctcaa atacttgaaa ccaggctttg aaaattcggc
900aaaaatcatc aaacctccga aagtatgaga gaggaaaatc aattatgatc ttcctaattg
960tgtcatgatt agcatagcgc atcaaagcac cttatcggtg caaaattaat ccctttcaaa
1020accattgtac ttcatgaatc ttgctacata tcttaaaaac gttgacgggc ttacagctta
1080ccagagatca aagcgagagg cagaacaaag ttcaaaatct gccgtgtata acaacaacaa
1140gtaattacag tagaaacacc ttactactca tcaatcacta ctactaacac caccacatac
1200gaatcatttc taccagatcc aaacaaagaa aaaaaaagag ggagagagaa ctaattaaaa
1260acacgaaatc cgaaccggtt caactaatca atcttcgcca caatctccag ctccagcgat
1320gaactgatga tcatggcggc caccaggtca ggtgtatgta ttcacaggtt tcgccgcgat
1380ttgacatgat tggaagtgga acgcaaactc acgcggccac tccggcgccg gcgccgatgc
1440aggagaagac gcggacgctg cgggccttgg ggagcgctac ccgcggcagc cgccggccag
15001381500DNAOryza sativaOs11g08230 location Chr114325545-4327044
138cgccgtatcc accttcacca gctcgatctc cgtcgttgat cgttgattga ttacgtacta
60cctgagtgat gatggacgtg gatcactgga atgggttgag aaaataaaaa tgatttatgt
120agtgtgtagg agtactcaac taacaattga acgaccagaa tttgacgaat tttatttaca
180tgtttagaga ccctgggtag agattcagta ataattaagt tcttgccata tttttgcatc
240aatttaatta ttttactaaa atgatttatg ttgttttatt ttcttggcac tatatttgca
300tcaaatataa ttaaacttag cttaactggc cagagcttat atatggctca gatttaattc
360ccctcttatc ttcagagcat taaaaaggga gtatcctaaa atacctatct cctaaaagat
420caatctgcct gtggctcaaa tctatataaa gtgagctccc ctcccactgt cccacatatc
480tatatagcta ccatggcact acagcttgca acacgctcaa ggaagtctct cgccatcgcc
540gtcgccgccg ccgtgccgct gctgatgtgc ctcttcctcg tcgccgccgc cgccgcggct
600gcatcgtcgg agacggcggt ggcgagctcc ccacagtacc agccgagcta tggtaatacg
660tactcgacgt gcttcgaggt ttcggcatgc gatgacaccg ggtgcgcgat caggtgccgc
720gacatgggtc acaaccctgc tggctcagcc tgctggacca gcaacgtcgc gaccatcttc
780tgctgctgcg gccgtggtcg tcctcctccg gttgcttgat ccatatgtat acataattac
840atatatggta tatgtataac tggataataa agcgtatgtg cgtgtcagcg tgagttggtg
900aagatgactt ataacatgaa acaggagtag cgctagtaat gagatgtgtg taaaaatgtt
960atggttcaat taattaatca aagtcgatct ttgcattgca tgctgataca tatctatcga
1020aatatatatg tactaagacc aagaatcaaa gtcgtactac tcgtatatgt acaaaataat
1080taattttagg caaattttgc tacaggacac cgtaattgtg tggttttagc ctagggacac
1140cgcaaaaaca aagtttgagg aaagacacta cgtaaccgtg gatatttgcg atggacaccg
1200caccgtctaa acgaaacttg ctatgctgac gtggcggtcg ggagcccctt ttttgacgcg
1260gtcggaccga aatgcccctg caccttacct cctcatattg cttgctggac actaaattgt
1320ttgctggaca ctaaatacat agatagatac atcgtacata tacatgacag atataccgac
1380atggactacg ttccttctag ccaccgctgc cggcgagctc caccgccgtc atatggttgt
1440tgcgcctggc gtgagaggta tatgcggcat tgcatcactg tccggccaac tttgaattaa
15001391500DNAOryza sativaOs01g64390 location Chr137374848-37376347
(reverse complement) 139tgtccagggc tccaggccgt gatcagcgct cgctgctacg
ctcagaccaa cacaggaaat 60tattacttgc tccctactac tacgtgcaag cagcgtaaag
ggggcaggcc actgtttgaa 120attcatccct ctcacctttt gaaacagccg cgcgtcgtgc
cgctgtagta gtgcagctgt 180gcaggtgcag tagctttggg tcaaaactca aaactcgaaa
gggagacagc accaaaaaga 240tactgcggct gcagccacac agggctagct gcttgacgca
gagagttcgt agcgacgttg 300ctcgtctggg catggcagtc gcgcctagat cggctcgtgc
acggcatgcg ttgcttgcct 360gggtacgcca ccacctgcgc agccagcgat catggccggc
gcggcgtccc acggcacgag 420agcgagcgcg cgcgcgcgcg gcactctcgt gaaggcgtac
gcgccatccg gccgcgcggc 480gcggcgcagg ccggtgcagc gtgcaggttg acccgcgaca
ggcgcacggc cgcacacgcg 540gtgcggcggg ccggcggcct ggatttgggc ggagttctcg
gtggccgtcg cttttcccgt 600gccagctgac gcgattacga gccgtacccg atcaaaaccg
gcgtgatcgc cggtccgatg 660catgcgtttt gttttcatgc cagatctttg gtaatcgtta
gtacgactct ctgagtccgc 720gagtacgctg gtttctgtta attgcagcct acagtatttt
ctttttcttt tttgcctcgg 780ggaagcgatg cgagcgatac gtgataattt caggtggcgc
agtgccgctg tgcatgtgtg 840aaatcatgca agattggcag gttgctagcg cgcacgtact
cctctctggt gtgatcggtt 900tcaggtgata gagacgatac atgggcatac cttgacacgg
agtgggctgg gatggcgcgg 960gtggtccatc ggctacagcc ttacatgctc ctcctatcaa
actctcgtta tgtccagttc 1020acttggccca tccttggttt cctctacgac tagtgctcct
ctagccagta gccactaccc 1080actgtttcca tattggcgca cgggtcatat aagatggttt
gttcctgttc acctgacggg 1140aaattctttc tcctaaaacg atatcgaacc atccttatcc
ggattggatt aacgagatcg 1200tcgtgctaag tgtgctaaca cacttgttat atgaccgtta
ttccgagttc gtcttgctgc 1260tgggagagtt gcagaattgt agtactccct tcgtttcatg
tgacaagacg ttttgacttt 1320agtcaaaatt aaattgcttt aaatttgatt aagttcgtaa
aaaaaagtac tacctccgtt 1380ttacaatgta agtcatttta gcattaacga gcatggtggc
tcgcgcaaat tgcgcggcta 1440gcatcattat attttctctc atataatagc atatatgttt
tctcattata ttattcaaat 15001401500DNAOryza sativaOs06g15760 location
Chr68954266-8955765 (reverse complement) 140ctccggcgag gcggtggact
cggactccga ctccgacgaa gtcgaagaga tgatcgaaag 60gtttgggagt cgcggcggcg
aggatgaggg gggtggattt ggtatattta cgggcgcggg 120tttgggcttt ttttcgcgaa
tgggccccgc ccggtttgac ttctgcgata gggatgcccc 180tcaaaaggta gaaaaatact
caaagaaata ctcctaactc cgtttctaaa tataagtgat 240atatatatac tactcaatcc
cttttagatt ggaagatgtt ttaactttga ccaaagtcaa 300actactctaa atttaactaa
ctttgtagaa aaaattagta atatttataa cactagcata 360gtttcattaa atctataatt
gaataaattt tcataatata tttgtcttgg gttaaaaata 420ttactatttt ttttacaaaa
ttagtcaaac ttagattagt ttaaatttga ccaaaatcaa 480aacgtcttgt aacctgaaac
ggagggagta tttcttttag ctggagaata ggtgaaacta 540tccctcttca aaattttaac
cagtcggatt tttataaaaa aaaatatttt caaatatttt 600agacaagcat attaccaaat
tatattgcat cttaaatata taaaagtatt gcgaagtggt 660gctatacttt ttggcgcaaa
aaaaaactca aatacttcac atcccttttt atataaggga 720aactttcttt tactgaaata
aataataagt gaaactattt atctttagat tttagccact 780cgtatttttt agactgatgc
cacttatttt tctaaacaag gttttcaaat attcaaaaca 840atattattct caaattataa
aagcttgaac ctttgtttat cgtaatacaa ccatgtaaga 900aatttttagt ggtgaaaacg
aaaaattgtc cagccaaata catgtcatct tcacatcttc 960tactagtttg atccaccttt
ggtgtttaat gttttcacaa aaggagtaag atactacgtc 1020gcacaatttt ggaatcagtt
gttggaaata atttattcta ttttggatgg cataatactt 1080tcagactttc attatagaat
ttactatatg atcatttttt ttatatttca gctggctatc 1140atttgaaggg atgggtgaat
ttatcccatg acattaatcc cccccctccc cctaaatttt 1200tataacttat cggctgtgtc
ttttatatac cacagatatg aatcagtttt catcattatt 1260aaaaaaaatc aagacaatta
ttttaaaaca atatttagcg cggagattcg tgctgggtcg 1320tccacgtatg cagcatgggt
gtcgctgtgg ggggtttggt gattgctggt cgttgtcgtt 1380tcgtggccgg ttgcagtttg
tagccaaggg tggtggcatc tgatatctgg ccacacactt 1440tggttatgga atgctgggtt
tgctgctggt gtggtgatca accggtgggt gattggtttg 15001411500DNAOryza
sativaOs12g37560 location Chr1223047433-23048932 (reverse
complement) 141tgacggcgtc tccttcttct tgcttcttgc ttcttcttct tcctcctccc
gatctggggt 60tgtaggaagc tactggtcgt gatcaatgga gcattccgcc atggatggat
tggatgggat 120ggggagaagg ggagggggag aagcgcgcgt tgctggcgat gttctctccg
cgtggggggt 180gggatgcgat gcgatgcgat gcggaggagg aggaagacga cgaggactcg
gctagtctgg 240agttaattga ttaattaatt gattaattag gagcaggaga agtgcaagca
cgacgagagt 300gaagggaaag gaaccgccgt ttcaaaaaag atggaatttt tgcggccaac
cccttctact 360agtgacgaat cacggtcatg tttgccacaa ctttcaggct gagttcgttt
ctttgatact 420ccattcgtac cgtaaaaaac cagcctaata ctagatgtga cacatcatag
tattacgaat 480ctggagatac ctctgtccag atttattgta ctcgaatatg tcacattcag
tcatatattc 540gttttttttg gacgggggag tatctaattc actcctcggc taatgattaa
ttaatcatgt 600actaatggat cactctgttt tccatgaaca cagcctcggg ttaggtctta
gatccacgac 660agatttaaat ttttaagttt tatttaaaat atgtgtaaaa tgatttatcc
aaatgttata 720gacataattg aggatctaaa tacatggatt tgtggagctt gaaatattca
gcttctaaaa 780atcttgaatt tggagctatg ctaaagagga cctacgtatc ctgcagttaa
cttattccaa 840aaagaaaaac aatatccaaa cctgatgctt aaaaaaacac actgaaaatt
aaaggcgtta 900tataaaaagt atatataaag tattaaactt gacaaaatgt gttggagaaa
ccatgcgtaa 960aactccagat attacctgga gacaaaaggg catgtagtca atgattgcaa
aacaaattct 1020ctggaagtaa cggaaataat tgaaagttat acatatccaa gtaggattta
ctacttacat 1080agccagccat tatccactga taattgctgc tattatcacc ataatcaacc
gaataatcag 1140ccaagaggtt atcccaacga taaatgactt tgatctctcc tttggtttaa
ctcaaatgaa 1200tgataatggc ttatcaaatg accagtcatc ttgactacaa aatcatatga
taattaacct 1260cattaattta tcactagtgg ctgataaaat gggatataga gaatgctttg
gaattcatga 1320tatttgctaa tttataggta aatttctaac aaacatatgg taaaataaac
ccccataata 1380atcgtactac caaactatac caaacaagta ctcttagcat gtatatatag
ccacaccgat 1440caactagaag actcaaaata ccaataggtt gtcggcatcg cctaaccgtg
atcagacatc 15001421500DNAOryza sativaOs03g17420 location
Chr39689781-9691280 (reverse complement) 142gtgttcgcac gacgataggg
ctctgacttc ttatggaaga tgaacatgat aaacgtagat 60gccttttcat atagtacgac
ctacctgaag acatgagatg agatttatta gacgttttgc 120tggtcaacac acaacacgtc
gatgactcgt gcgacgcaaa tgcaaatcca gatgagttag 180tggaccctca cacttataca
taggagtagg tgtcatctga gtcaatatac tagcaagagt 240gagtttgaaa gagatgttgg
gattgagctt ttttttagca catataaaac gagataagtc 300attactatat gattaattaa
atattagcta ttgcaatttg acttctatgt aaaaattttt 360tttcaaaaac atatacatta
gtttgcaaag cgatgaaata gtttacccaa cttctcttta 420gaactcagcc taaataaatc
taacccctaa atatgctaaa tgtgccagcc ctagtccaaa 480atttaatgag acctaccata
agaatgttga catgacatct tcaccaatcc tacaaaccct 540taaaaagtta aaagtttgaa
gaaagttgga agtttagaaa aaaagttaga agtttatgtg 600tgtagaaaag tttttgatgt
gatatgatgt gatggaaagt tgggaatatg ggggaaacta 660aatacggcct aacaagtatg
gcacacacac gctttatttt tacctttttt cttcttttta 720ttctgtcttt cattctcctt
ttctatttct ttgcttctat gttcttcctc aagcgagcct 780tccttgagct caagttgttg
ccattgagct catggtgggc tggtgggcat agtcgtgcac 840tcacactaag caactatcgt
taagctttcc atggctgctg taacatcccg gcccagggct 900taataggatt aatataagac
ttgtgcacac gagtgaggac aaagtgtgca gaaaagactt 960gtgttggtct gtgagggcag
tctatgacct atgaaagctg ccagatgtaa gcggaccact 1020tcttttctgg aagccgatgt
ccaaagaact ttagggttaa gcgtgcttgg cctggagcaa 1080tttgggatgg gtgaccgacc
ggaaaattct tcccaggtgc acacgagtga ggacaaagtg 1140tgcatgaaag ctgccagatg
taagcgggcc cagcttggga gaggcgggac gttacagctg 1200cgatgcttaa gcgtccctat
cggccgccac catcgagctc gtcctaccct tggtcaccac 1260cactcctcta ccatcactga
gctcctatgc atccacggcc aaaacaaaaa ccctatcctt 1320acgtatctcc tctttcacca
accgaaaccc accctagcct ccattgcttc agtcaccgcc 1380attaaacaat attagatcga
aatcctaatt cttcgccagc acaaaatgac taaattcatt 1440agtgttgctt ctcatgttcc
aaaattaaac atgtatacga gttcattgta cccggtaatc 15001431500DNAOryza
sativaOs04g51000 location Chr430185779-30187278 143gcgaggatgg ctactagctt
gctgtccttg cggtcggccg ggctcggttt ctccggttgt 60ttcttgtgtt tcgcccacac
acctctgtgc gtcgcgttgg gctcgttata tagcggccac 120tgtagtgttg ggctatagtt
gctgcacgcg gttgacttga caaatctcca tagctcgttg 180cattggtccg gatcgatgta
tctgcattca cgctagcttt tggtttttgt ccaatacttg 240gaggaaggga gcgagctacc
gatcgatact acgtgaaaac gacctgtcct gtagaaagct 300gcatgcgtcg ctagagcaca
cacgatttga tagtctagat tctagtaagc cctattacgt 360gccggtacat aaaaagtagg
cacagtaagc cttataggag taatacaccc acacattgtg 420ttgtcctgtc acggcgtacg
tgcatttgta atgtggcgca cgtctcaagt gtccttcgat 480cttttcgtcc cgtctctcct
tggaggtagc aacggcgtca ctgttcctct tccgaagaaa 540aaagatactc ctgcatgcgt
actgttgttg cgtacacggt gaacacgggt gaggccgtag 600ccttgtgccc ttaattatcg
tacgtggcgg ttgtccatgt aatcgtgtga gcggtggggg 660cagaagatta gcgtgtcatt
ctgcagattt tccccacgct agggcaactc aaaaatgttt 720ttgattccga gggcgaaaat
atacttttta agaattattt taccattctg gataattaat 780gtatcgagag acaccgtatt
tttttaatat aaaacttaat acttttagat actgtgtatt 840ttgagataaa caaaagagta
aattgcatca gcggtacacg aacttgtcag gttggtgcaa 900tctagtacat gaacttctaa
aacgctcgtt tctgtgcacg agcttgtttg atgcgtgcga 960ataatgtcaa aatcgcactg
caaggttaat cttgttgatt ctgtggttga tttaaactct 1020atgtaaacat tagtctaaga
aggataaaca tataataagg cataatgatt gtataaaaaa 1080ataaaaattt gagtaatgat
gcaagggaca gaaacaaaga aagataaagt ggaatctaag 1140aaaaatcggt cttgtcgcac
cattttagtc ttagtttgca cataccagac aaattcatgc 1200accaaaacga gacgagcatt
ttataagttc atacactaaa ttgcacgcac ctgacaaatt 1260tatgtaccgc taatataatt
tactctaaaa aatgtagtgt aaaatttctc ttcttttaaa 1320gtctcactat ggtgcattca
ttggtcttat ttgattggag gatttagatc ttttttacaa 1380ttatttcaat taacggtagt
agaaatatgg agtgatattt attgtaaaaa agaataatgt 1440gcatggtgta taattgtcat
ttagcaatga ttaaatattt gtttctcttt ctggattttt 15001441500DNAOryza
sativaOs01g01960 location Chr1522445-523944 144gacctcgagg aactcctcgt
atttctccct cttgtcctgg aacttatcct tgacggcctt 60gaggtagacg agcgcatcgt
tcgtggtgag cttctggccg gccgttgcgc cgccggcggg 120cggctgagca ggcggagcag
cggcggcctg aggcggcggc gctgcagcgg aggcaggccc 180cagaggcatg tgctgcggct
gcgccgtcct acaccacgag aggaggggca aacagttaga 240tcaaaacgca aaacaaaatc
gcaccctaaa aaacacctcc tttttttttg cgacgtagag 300ttagaaattg gaacacaaaa
tttgacggtg aagaagagag attccccaaa ggaaacccga 360attcctcccg cagatggaaa
taatatatat aattaattat tcactccctc gcgggctgag 420ggtgagggaa acgaaaccga
agtgaagttt attttgaaaa ataaaacaaa cgaaataggc 480aggaacgcct ggaagggcga
aggcgatgag caccgaggaa gccaaggatg gaggaggcga 540ggagaaaagc actcacggat
cggatcgacc gacgttgggg cgcttgagtt gggagcccat 600gagcgcgtcg tccctggcgc
gcttcatccc cccatacaca gactcccttc ctcctcctcc 660ccctcctccc ttcacccaac
caaaccaccg ccgcctccta cagaacaacc tcccggtcgc 720cgccgccgcc gccgccgccg
atcctctcaa acctcggaag acgtacaccg gcgccggatc 780tgcccgccgc tggctcctgc
gagacccccc gagcgcggcg cgaagagggc gcagcgcgag 840gcgaaggcga cgcgcgggga
ggagaggcgg ctaatcgcct cggcgagacg cgagacgcga 900gagggaagga tgggtgagga
acgaggcaag ggcgaggcga agaagaagaa gagaagcgag 960gcctcctctt ctctttaaca
caacgaagag aagagagcgc atcacaaccc atgtacaccc 1020accggtgcca ccacgctgcc
ccgcgcccgt cgcgaaccac atcccgcccg cacatcccca 1080gataagggac acgtggacgt
accggatcga ccgccctagg gtgcgaaatg gttactggcg 1140gtgatcgcac cggtgggagg
gttagcttgt ttcgtgggta aacacaacct acccattgga 1200tttggggatt ccagtgagcg
gtctggatta gcgcggggtt acttcgagat tagtgccccc 1260gggcccacct gtcagtcgga
tgagtgcctc cgatctcagg tttaacctag ctccgctagg 1320gcggcgtcgg ttcgtatatc
cgcctcatgc gtgtgtgtct tctggctcag gagacgtatt 1380tccgtgggat ccgtatatac
atgggcttca tttggccatt tgtcacgatc cctagcttcc 1440agagagggct accgctggat
cttcagtttc atctgcagta tcaaactatt aataaaaaag 15001451500DNAOryza
sativaOs05g04990 location Chr52418356-2419855 145tgataacgat ggtgcgcctt
cgtgatcgat cgagagcgtg aggagcaacg cggttcaatt 60tatagacgat gcagatcaag
ctggtgccaa gaagtggcgg gaggttgatg acgatcgatc 120gatccgagga agaagaagaa
gaagaaatgt aagtggtgat ggtggtgatc gatcgagttg 180agttgagagg cggcatgcgc
gcatgcatga ggatgagggg gaggcaatgg taatgattgt 240gagcattatg gggaggggag
aggaggatct tggtgattaa tgaacaacgt taatggtgga 300agtggtggca gtggggattt
cgaatgagat ttttcttttt aacgattagc actagacaac 360acgactatga gaaataccca
agggtcatct ggctagctct acaaggtggt aggccagatg 420atctgggttc aaagcctcgc
cccttttaat tatttgatat tagtcttttt ctaatattca 480tgtcttttac tagacagtac
gattattcat cgaatagaat atgaaaaaat tacaagatta 540attagagacc tagaagacaa
tagctaggaa agaagaaaaa aaatccacca ccacgcaccg 600acgtcatcta cacacagcca
aggagaaggt ttagcaccgg accagtcggc gctaagcgtg 660accgactgcc gctaaaccaa
caagggtatg ggaatgaccg ctaagattgc ccaagtccaa 720gctataacag ccgagcatca
aacacaactc tagccacatc tataaaattg aggggaagag 780gatgagagag aagaattgaa
ccgctctcta cgctaggccc tctaacgtgg ccaatttgtt 840taaactaggg cgccaccaca
agaggatgag acatctaaag tgagcttgca ccagttatct 900atgataagat ttgtcaaggg
ggctttctag atgacgtctc cagggacagg agcgacaatg 960acactgccgc caccatccgt
cgaggtctta aggagaacta agacaagatt ttcactcgat 1020aaccctacaa gaggagaggg
gatggctcga caacaccccg aagaagtaag atggcgcctg 1080aagcgccagc caagaccggg
ctgggtttac acccgccgtc ccgccacctg ccgatcgaaa 1140gctgtgctcc attgctacaa
ccaccatcca tcctccgtcg cgcgtgggca ccgttgcacc 1200gccggccccc gccggccaac
ctccatgcgg cctcccactc tttgcaccgg acagttgtca 1260ggccatcacc gtcgccgacc
gccggccttt gccgctccgt cacccacgcc tgccacaagc 1320cgtcggccta cgtcgcccgt
gcccgtcgca agccgctggc ctctgtcgct gtctcctccg 1380gtctctagat tcggcagcgg
ctgtgccgga tctaggcgcg ggggagcggg ggtggatgtg 1440gaggtggggg aggaaggggg
gcagatctgg cagcggtggt gccggggaca ctatctcgag 15001461500DNAOryza
sativaOs02g44970 location Chr227240720-27242219 146ggttgaattc cacagggaaa
gctcatcatt tacaattttc tggaaaacaa gtcttgactc 60agacagggcc cctaaattag
cataacaact cacaatctta gagccaagaa tcacatccca 120gcacaatcca tgtgtgaaaa
catgacaatg gatcttcttt agagacctaa tatctgcaca 180gccttggaac aatggagcaa
acttgtcaaa attaagtatc ttattagagc atgaatttgc 240atcggcagtg gatgctgata
gtctccaacg cagcttcagc actgcaattt tatcaaactt 300tgaactagtg tttccagcta
aattcccctc aaaagaattg gtagcatcat catggttgtt 360gacgcaagaa aaacaaaaat
aggtaactaa ttgactccga attcatagac atacgatgac 420agttcagtta aatgagccta
caatataata gtcccaagca cacatgtgtg agtcatgtgc 480aatgattaca tcatgtgtgc
ttgggactat tatattgtag actcatttaa ctgaactgtc 540caattgtggt tggtgacatg
gttcaggagt ctaacggtgt accggaaaca gatatagtgt 600atatctggaa gtggagtcta
taaataggtc cctaccctcc agcctcataa tgcattgtga 660gcggaacaac ggaaaaggaa
tagagggtag gccggataga cccacgtaaa cctaacaata 720tttgcttttc ttgcacctcc
aaggatagac tattcaacta aacctgatca acagcggaac 780acatacaatc agtgtacttt
agtacaacag cactaaactt gatagcagat cacattaaat 840taagcgataa tcccagttat
ggcatacaca gcagaggagt gaagccacaa tctctccagg 900acattgccac aggtaatgaa
tatgaaatac taccgcaatc tcgccggtgc taccaaataa 960accaccacct aatcagaaca
aatagccaca tcatcaagca gtcatcatgt aaaaacgaga 1020aaatgggaaa actgaccacc
agccgaaatc tttttcagag gttcattccg tgaaagaacc 1080ggagtaggag gaggaggcct
cggggtggcc gccggcaagc cgacctccgg agggacggcg 1140ccaggaacag tggaggaggg
gcgcgcgcgt gggggaggcg cgggtggtgc ggccgtgcga 1200gggggcggcg ctgccgctgg
ctagtggaga catcggagtt cggagcgagg agaggtggcg 1260tggcgagacg gtggggaatc
gcgtgggcct ttgcgcgtcc cggagaaacg gccgggtcga 1320aagcccacac taccattcgg
cttgggacta gctatccatc acgcgacaga tttttttttt 1380tccgaatcac acagatttta
tttttttttc aaatcacaca gattttagat taattgaatt 1440aagctcattt gatcacttga
ttagaagagt tatcaacata tattgcatat attttattta 15001471500DNAOryza
sativaOs01g25530 location Chr114470301-14471800 (reverse complement)
147ggttgtagat gccagaaaaa tgaatatgag taactaattg gcttcaaatt cacaaacatg
60ttttgctttt attgcacctc caaggataga ctattcacct gaacccgatc aacagtaggg
120gacatataat cagtgagctg tagtacagca acacaaaaaa tgatagagaa tcacattaaa
180attaagccag aattccagtt atgatgtaca caacaaacaa gtgaagtcca ggggcggatc
240caacttggga catgggggtt cagttgaacc cccaaacttt tgggtgaaca aacaaactgt
300tattacctga attatttaag agaggtttaa ggctatcaaa ttaaggagga ggagaagatc
360tagaagagaa gctagtaggg ttcacgaacg caagcaaatt ccagtcccgg tggggtggga
420gagagagaga gatgaatcag aaagaggacg caccaggatc ccctcccctc cgctgcgatg
480gagatcgccg cccggccctc ccctgttcag ccgcgactcg tcttggcggc ggcgcaacga
540ggcaaggagc ggaggagaca acgagaggta ctcgactact ctgcccttat ctcaatattt
600ttacttacca ctctctacgt cctaatagtt ttacaaggtt cacatccaac atttaacttt
660ttatcttatt taaaaatttg aaaatttttt aaaaacggac ggtcaaaagt tcgacacgga
720ttttcacggc tacacttatt agggacaagg tagtatttta aaactagcta gtgttatttc
780tttatataac tggtgaccca tgtatcatca atacttagga aaaaattaaa cgattgtgtc
840gacaacaaat tcctcatacc accagcaacc gctaactgcc actctacgca ccgccatcac
900tgctctttct ccatgaatca ccatcgttct tctcctttgc ctttacatgc gcagccttaa
960cctctagtaa ccgacaagtc tttttctcct cccctatcgt gatgcgcact attgttatcg
1020ttgccattga ggtcggagat gtcgggcttg cagggacttt ggaaggcagg agctgacccg
1080acgaagaacg aaggcgctat atccgggctc tttcttgcta gatctatcgg ttatccaact
1140gtcgtatgga taaagaaaat ggagagagag aaagagagag agttaatgtg cgaagtgatt
1200actgagtcac cggacttgga gatggataga atatggtggt ctaaattaaa atgtaaagtt
1260agctattgtg taaatgaaaa tggttaaatg ataccattat atgaaaactt ttaagaattt
1320ttaagaatac atagtatgga ttgtaattta cttcctttgt caaaaaaacg aatgtagaac
1380tggtgtggca cattctagta caacaaatct gaacatatgt atgtctagat tcgttttact
1440aggatgtgtc acatccaatc ctaggttagt tttttatggg acgaagggag tacttttttt
15001481500DNAOryza sativaOs03g30650 location Chr317474886-17476385
148ctccgactct cttgtcttct ctgcgtgtgc gcgcgtgttc tgctctgtgt ttgtgtggga
60gaaaggttcc ggcgttcagc gaggtttgga ggtaggagtg gggccctatt tatgcactga
120ttcgaatact actctgtatt gtatttacgg tggctaattg ggattttctt ggggtatttt
180tataacgtac ttctgtattt tacggtgcct aataattggg atttgtttga ggtaggggtg
240aaaacggtac ggaaacagac ggaaaccatc tttattgttt ttgtttttat tttatttttg
300gaatcggaaa ccccagatac gaaaacggaa tcgaatatta tcgaaaccga aaacggagcg
360aaaacaaacc ggcgcgaata cggtaacgaa aatttatcgg aataaaaaac ccctcaaact
420gataaatcca attgaataaa ttgtctatgt ttaaaagagt aatgttgttg ataaatccca
480atataggaaa aaaatattct tatgtaattt tagattcgca taacaaatat ttttttaaaa
540aaataacagc caacatcatt gtattcacta tttagtgctt aaaaaaatac atggtatttc
600agtcacgaaa tttttggtaa ttccggaaag tttccgaccg attccgagtt ccgatggaaa
660ctgcccttat catttccgat tccgtttccg agaaaatatt tccgaattcg tttctgtttc
720tgaaaaattc cgaccgacag attccgtttt cgaaaatagg tctggaatcc ggaaagattc
780cgtaccgttt tcacccctag tttgaggtga taagctttga gatttacctc aagggtccca
840cgagcaggtg tagattctag aaacgggcag cgcccagggg ggggggttga aaagttcgtt
900gaaggctgtc agaacaccat ggaattgagg aattgcgtca aactggaatg aacatgatga
960acacccaagt actggcatga ttagtccact tattattcaa agccaaaatg gcgtaatggc
1020gtacgtgaat ggcgttgcaa aatcatccgg actgcgactc gcgcgcggaa gcctctgcct
1080ccgcggcgcc ccaacgccgc acgaggatgt tctttttctt gacgtcgtgc ctcatttcga
1140gcgcagcgag ttagccctgt tttcccgatc aaaaactttt tatcatgtca catcgaatat
1200ttggatacat gtatagtcta ttaaatatca aatatagaca agaaaaaacc taattacata
1260gattgcatgt agattacgag atgaatattt taatcataat tatgccatga tttggcaatg
1320tggtgctact gtaaacattt gctaatgaca gattaattag gcttaataaa ttcgtctcgc
1380agtttacagg cggaatctgt aatttgtttt cttattagac tacgtttaat acttcaaatg
1440tgtgtctgta tccttcaaaa acattacacc caaagaacta aacacaccct ttgtattgtg
15001491500DNAOryza sativaOs01g64910 location Chr137684066-37685565
149ggcagcatgc cttgccttcg atagttcgat ctctagttag cggcaggcgc agtgcaggac
60agtgaactga ttgcccaagt tcttgcttgc tgaggactgg atgggagaag aatgtagtac
120tagtacttcg gaggaaaaga gccaaagaaa cgctcgtcat ggctgacgaa acagttgaag
180gaatcaggtg attgatcgtg ttcgtgtatg atacacggtg tatctggaat tctggatccg
240gctccgactc cggctcctgt atctgtatat gtcaaaaact ggtgtaaacg agatccctcg
300acggtttgaa agacagtttt ggctgacgtg atgcttacgt ttttttatta tttttccggg
360atcaaattgt cgcatatgcg ccatgtcaat gttacgtggg acgaagtcct aatcaaacca
420gccacgcaaa cgtcacatca gtcaaaatcg cctttcaaac cgtcgaggga cttcgtttgc
480acagattttg acagttcagg gaccggttgc atctgatttt tggtttctaa gaacgaaaat
540cggatttggt gtaaagttaa gggatctgaa atgaacttat tcatttctaa tagggcacat
600cgccttcatt tcaaatgggc ctccatgtgc caggcccata tttcgattcg agtgtggcct
660ccatggacca attgagaaat aagttcactt taggtcactc gtattgttgg agagtctaat
720attcattcca ggaccaatta aagtggccca tattgcggac gctgaagccc aaaggagttg
780ctcctatggt gcggttggaa tggtggatgg ctgaagatgc ctcggaggta gagtagtatt
840ctctcattcc caaaataaac tggagtaaga gcatctccaa tagatgacta aaattaaact
900cccaaaaatc atgtattggg gacagccaaa aacatattta gcctaaaata cacccccttc
960tccaagagag gactaaaatt tgggagcgct tctagttgcc caatatttgg ttcaggttgg
1020tcctggtttt ggaggtggct aaattttggg accatgctta ggagtctgtt ggagggctga
1080ttttcaccaa attcctaaaa tttatgtttt agtaacctgt ttagcattct cttggagatg
1140ccctaactac catctttaag ggagtatcta aaacagtgat aagttttttt ttaaaaaaaa
1200tttagatata actatagtat aattgatacg taattacatt gtaactatat tgtaacataa
1260ctatgatatt ggcatagttt ggtggtttag tggtacgtga gcattcgaca gatcgtgggt
1320tctcaatcct tgaccactgc atgcattggt taattgttac tccctccgtc cataaaaaaa
1380acaacctagt actgacacat cctaatacta tgaatctaga catacatctg tccggattcg
1440ttgtactaga atgtgtcaca tctagttcta gaattgtttt ttaatgggac ggagggagta
15001501500DNAOryza sativaOs07g26810 location Chr715496040-15497539
150gaatgatcat gtcgcgaaga tctacggggc gacggcgagg cggtgaacga ggtcgttctg
60ctcggcgctc cctatgatgt tcgggaggcg agtggacaga tcgctcgtcg tgacccctgc
120tcctgcgatt agatcccacg ccggcgatgc tgaggcttgg ctgatgatag tcatgagatc
180ggagatgggg gactcggctg ccagtcggcg tgcggacgct tccggagtta actgggaccg
240aagcagcaat catggtcttg gtctggttca agatgttgac cacgtgatca gattgactga
300gcttgttctc cttgaggagg gtgtcgagga gggtgatcgc agcaacagca ttctgttgtg
360gggtgcggaa aaccggtgtc ccctcaacat cttgctggcc gataagatct cgcgctcggc
420gccctgcatc taaggctcat tgccgtcggt cttcagcctc cttagcggcc cgctcgcgct
480cctgttgctc acgccgcagc cgttcttgct cctgtgcttg acgctcctcc tccagtcggc
540gtcgttcggc ttcacgtcgc gcttcggctt ctcgggcttg gcgttgctcc tccgtctcgt
600tatttactcg atgagaggta ataccctact cctgtttggg gatttaaatc caccgggtgt
660agtatagatc tgacgatcat atgtgctcat gcccctagag ggcctcctgc ccaccttata
720taggatgggg ggcaggatta caagatagaa accttaacca atatagtatc ggtttcctaa
780atttatttta caatattatc aaatcaggac tttaggccgc tccataatat aaaaggaaac
840gtaataccca agtcatgatc tgttacatat tccacagata taagctatcc cctatgacta
900gtcggataac catgccgtgt gggtatgggg tacccataat ctccacagta gcccctgaga
960ccttcacagt cgaaaagata atcttttctc gaactagatt actccaaagc cgagtgcttc
1020aatcatcttc gccatgatct cccgagtact tttaccaaat atgaagactg tggagagctg
1080aaaaataaag tcaggtgcaa cgactagatg catctaatag gtgtagcccc cgactatgtg
1140gttggctgaa caaactaagc atatagtcaa ggaataaatg attcaaccaa ccgagcggtt
1200ttgataattg taatcaacca agtgacttga tatcaatata tagcatatgc ggtgtgaatc
1260ccccaaataa catgatccaa gtaatatacc gactgctttg taaaaattga tgcgagcaac
1320ttaaagttga ctacatcatt gaaaattcac atagcaaaac agctataaag aagcttgaat
1380gttgcaaata aagtattggg catacgccat gcgcacactg ctttaacata tgtgcttaag
1440ttactaaaag acacatggtt tcaccacacc tggaaatata tggcatatgt ggtgtaaaat
15001511500DNAOryza sativaOs07g26820 location Chr715515227-15516726
(reverse complement) 151agctgatgta gcgggtaggg gagttgatgt tggcgccgtc
ggctgaattg tggacgcaac 60ggaggaaacc gcttgaacag atggctgaac atcggtgatc
aaagatcgat aattcgggaa 120gacacctcct tgcaagtaaa ctgggcctgt agcctgatgc
tcggctatcg aaccatcggc 180tattgtcttc atcatgtttg acaatgtatt gactaaaacc
ccggactgat tgatcagagc 240atgatgcaca gcgtaatcaa ctcgatcttg aaagttgttg
aaaaaatctt gtgctagctc 300attattctga ttaagatttc cttcttgcgg cccttgagca
ccatcacctt gagccccacg 360cgctcctcgc agattgtcgc cttgatcgct tggggagccg
tcggtagcac ctttagtact 420cggctgagcg tgtatcggct aatactccga taccactcta
tatcaggata ttaaagcagg 480tataatatat caaacaaaag cctaacatat ttagatacag
caatatcttg atataaaggg 540cagatttagc atatcaatga gacatataga ataaatatgg
ctaaatcaga tacgatcggc 600tgaaactcca atgctattct aatcggcaac tagaaggcag
gctagagatc gatattctaa 660gcacgactta atagatcaaa ctcaacttat gcagcattaa
gtatgaaaag aagaacgata 720tctagacaat caagccgcta gcagttccat agaatggtgg
atatcttata tgatctagat 780caacgtcaag atctaaccta atcggctgcc ttctgcgtac
agatattggc cgatagtaga 840ttagatagcg atattgttag agattatata agatatatga
taactcgacg aattacataa 900acaagattag agtgtcatga agatggaagc actaatcccg
agaacgcaag ccgtcataac 960aagttttacc tcttgttgaa tattgaaatc gatgcagctc
aacccgaaag caagaacttg 1020tcgaaacaaa actaaagcaa aaagggtggc gatgcgccga
gattgtattg gacgtgtgtg 1080ttaaaaaatt acatagggcc cggggtctat ttatacccga
gaattacaag atatgcccat 1140accggacacg accattatct ctaacaaact ccaagatacc
ataagtcttt gcggcagatt 1200tttgcccaca cttatctata aggaatttac ataaaatatc
ctaattaata gatacaattg 1260ccttcccagg actctatcca tgtatggcaa tcatcttgaa
gtacattaac gtgaacccga 1320tgtcgcgatc aagccgtatt gtcggaatcg gctgtatcgg
cttatttagc tcgactcaga 1380cccagccgat cttaaccgta gccgatctgg actccagccg
attcctgctc agccgattcc 1440tactctgttt ccgaactcga tctccgcctc cgactccgct
ttgatcaaat cctctttcct 15001521500DNAOryza sativaOs09g11220 location
Chr96225987-6227486 152cgggctagct acccccttat caagagcctc cacctcatcg
gctcatccca ttctcttcct 60cacctccccc tctcttctct ttcgtgatcc ctcttcctcc
tcctcctctc tctctctctc 120tctctcacac acaccaattc cacctcaaaa cgcatgaata
tagatgatgg gttcatttgt 180atggccgtat gaagtttaca atgatccctg tattttctat
cttccttatc atccttatcc 240aaagctcttg ctcgatgtct ccgattgtgt tcttcaaaaa
cacctactta ttgtttgatt 300gctttggtct tttgtgaagc aaaataaaaa agagcttgaa
gtggataaaa tttttcccat 360gccttcccat gaaatcagta gcgaaggaat ctttccttct
atttttcgta cggctggtgt 420gttccatctc ctccagatcc tcggacgtag cactcgctac
gtcagaggga catccggcgc 480agctgcgtcg ttcataccgc atcggatgaa cgcacgcagt
gaggcggcca aacgggctca 540aactcggtcc aagtcactga aataggacgg ctcatttgaa
cagaaggctc gtttccttag 600ccagaaaaat cggtgtcgct cccaagttct ccaggacatc
agccggcggc cgctcatcgt 660ctcctccggc tcaggcgcta ttggagccga tctcggacaa
agcccctccc ctcgtcgccg 720ccctcctcac catccctcga ctcctcacgt ggcgcgatgt
cacgccctga agttttcccc 780ctttttcttg ctttaaaaat ttgtttaata aattgcctca
agaaataatt tgattaacct 840agagctaatt ccttaattaa taaatgcaat caataattgg
aaatggcatt gtgggatttt 900tcttgggttc cacttgtcac atttcattaa cgggattttt
agtagaattt tcatagccct 960ataaatagtt ttaaccaata aaaatcaatt atccgcaata
ttctaatccc aggaaaatcc 1020ttttcttttt cctctttttt cttttcctcc tttttcctct
tcttgggcca tcggcccact 1080tggctcctac gctgcccgct cggctgggcc ggcccacgcc
ccatccctcc tctctgggac 1140gccgataggt ggggcccacc tgtcaagtcg tcccctacct
ctagccgggc agcaaccgcc 1200gctgaaaccg cccacgccac caccgttccc gctctcctcc
acgccaccac tccacaccag 1260tgccccacgc ccacgtcgcc cgcccactaa cccgcttctc
ccgctcgtgc gtgcgcccgt 1320gaggggcagg attcaatttg aatccccccc tctctctctc
ccccacctcc ccacgtcgcc 1380agccaaatcg ggccctttcc cggccgtgtc cgcctctccc
aaacccctat atagcttccc 1440tgcgtcctcc tctccatttt tcccctttcc acctccctct
cccgtgacct ctatcgcacc 15001531500DNAOryza sativaOs04g21800 location
Chr412352409-12353908 153ggtgacgggt ttatgacgac agcgggttgc aggtgcgatg
atcgtccttc ctctcctctc 60tcctccactt tcccccggtt gcttggctag tggattgtcg
ggtcccaaaa ctaatgattc 120ggtaaccgac atgtttagac tgtatcaagc cctggatcag
tagattgata caggttcaac 180aatctggatc tttattgtat acattttaat aaggctccaa
aggagatatt gttattacca 240aatatatggg tatttacaaa cttgtggcca actaatacaa
cagaagctat agaatgaacc 300aactctaatg ttttgcgtag agttaaacca tcaatctaat
ctatacttga aaactaaact 360aaattagaat gagactaact tatatgattg aactcccagc
ttcggcaaaa actccgaaat 420gactctgaaa aaggtggggt tgaagcaagg gtgagtacaa
cgtactcagc aagctattat 480attcaatatg aatgtatgaa atagtagcat ttgagtaggg
ctagatttac ttgcagaaag 540cagagaatgc agaagaagtg agcctgtaat gattttaatg
caacagtatt taataaagtt 600tggaattaag tttctaacca aataccatat gagtcccaat
gctcaaatcc atgagcacgg 660ctattcgaat agattcgttt tcactttact gcagtaaatg
tatgctttac ccatagccca 720cgacgtgacg ataatcatca gctttagtca tggcccagca
ttaggttatt aacaatagtg 780gcacctgttt catgaactct agtccccatg cgctctgaac
gtacgttatc agcagcgtga 840ggagttctgg cgttcctggg gtatttagag aggactgatt
gtatgacacc atatcatcgc 900aatcagggtt tacaaacagt tcgtgatatc acaatttatc
tcaaatattt acctatgcct 960cggtaaatat cacaatattg ccctgctcgg cataagtttt
cctcctgcgc gaggacttaa 1020acaagaacca ctatacagag gtaccacctt gttaaataac
ataataactt ggtctgtccc 1080catcctagaa ctgtggtcgt actcgtttgt tcttcataag
tacttggcag tcttatgtcg 1140gttggaacag tactagccac ccggaaaatc aaccatttct
accgtaccgt tcaaatctaa 1200gtttattata tttgtatgca gtctaactag gcatgactaa
gcaaagctag catatatctg 1260gtttgctata tgttcatgat atgcattcaa aatcatgaag
agctaatgca tagaacagaa 1320ataaagaata tagggcattt atgctcaaag gagaggaata
ataacttgcc ttgctccaat 1380gcaaataaat ccgagatagg caacctgatt atctgatcct
tgaaatatca caagttgcca 1440tctaaatata atagactcta ctggagaaga ggaagaacca
aattcaataa aaatcatgaa 15001541500DNAOryza sativaOs10g23840 location
Chr1012223468-12224967 (reverse complement) 154gacctcgccc cagtgggcat
gccaaaaacc tcacacccga cacggcacca ctttacttac 60caagctcaag aacacccaac
gtaacgagca gacgcgcagg gcagatcaaa tatcaccggt 120acgaaatctg agacaggata
gctcatgcgg attatgcgtc taaacagctc aacttgcaac 180gacaagctga tacgcgagct
cagctcatag cgataacctt ttttttacgg ggagtaccga 240tagctgatgc ggcgtcgggg
actcggggca acgcaacctg atgcgaacaa gcgaccgaga 300aagagaaaca tcacatacca
aaattacccc taatatatat taagagatta ctaatctacc 360ctttaaaata tacggttgag
attaattcta cttgcagtag aatactgcaa gtagaaagca 420ccggagcgtc accttaaagc
caaataatgc cttctgaaat ctcatgtaca actagaagta 480gaaaatttct agtggaaatc
ttgcatccgt ggttctatga atcttgttat ctacagtgat 540caatcgatca atcggataac
ctgttctttg atgaatcttc ttgagttttt atcggatgga 600atttgctggg attgctttga
attcgtgcac aacgccgtcg atctcgatga ggctgcagcc 660ggggaccttg tcgatctcgt
tcctcctcat ggcgtcgagt tgctggagcc cctcctcgat 720gagtccggca tgacagcacg
ccgtgaggac gccgaggagg gtcacctcgt tcggcgtcac 780gccggctcgt tgcatgctgg
cgaatagaga cagggcatcc tcgccgcggc cgtgcattgc 840gagacccagg atcatggata
tgtaggtata ccggcgagac ccagcgacca gcggccgcag 900caagcctcgt accggccagc
gttcaacagc cttctcgcgc acggccacat cctgctcgcg 960ctccgcaccg ccgttgccgg
ccttgcgcgc agccgcctcc tgctcacgct ccacgccgcc 1020gtcctcgcgc tccctccccg
accgtgtccg catccgcgcc attggtcgct cctggcgcgc 1080ggtcgccgct acccgctccc
ttgccgtgga caacctcgtc ggcggcggct gctccctgat 1140attttttttt tttggtgtga
ttcaggtgga gaaagatgga gccaggggca atcttgccat 1200ttcgaaaaat ttctcacctc
ttttgacccg gatattataa aatattgtct tcggtgtcct 1260atagctaatt acataatttt
tgtagtgtcc atcagccgtg tctcgttttt gcggtgtctc 1320tcagccaatt acacgttttt
tgagtgtcct atagcaaatt ttgccttctt cgaacggcgg 1380gaagaagttt gctttgtaat
ctatatagtc aacacatact aagtttgatc gtaatcattg 1440cttacacagg atttggtcac
atttatgaaa atgacaatat agccatattt gttaaacaaa 15001551500DNAOryza
sativaOs08g13850 location Chr88268007-8269506 (reverse complement)
155gacctcgccc cagcgggcat gccaaaaacc tcacacccga cacggcacca ctttacttac
60caagctcaag agcacccaac gtaacgagca gacgcgcagg gcagatcaaa tatcaccagt
120acgaaatcga gatgggatag ctcatgcgga ttatgcgtct aaacagctca actcgcaacg
180acaagctgat acgcgagctc agctcatagc gataaccttt tttttacggg gaataccgat
240agctgatgcg gcgtcggggg ctcggggcaa cgcaacctga tgcgaacaag cgaccgagaa
300agagaaacat cgcatatcaa aattacccct aatctgtatt aagagattac taatataccc
360tttaaaatat acggttgaga ttaattctac ttgcagtaga atactgcaag tagaaagcac
420tggagcgtca ccctagttgt aaatgggcac cattctttga ttctctcgtg atgcatatat
480acattacgaa gaatgtcaaa atataactac tggtgtgaac atgaagttat ggttgtttgg
540tttcattcat caaggttcaa atctttatgt ccatatacac atctcgcatg tgtattttaa
600ttggtgcaga gaggcggtca ttctatttct cttgttaaaa aaaaaaatca aactataacc
660atgtgttcgc catgctttat accttccata caaaacttgg gtcataaatg tgagctgcca
720aagtgatctt aatggtttag aagtaagttg cttcctaaca tggctccatt atagaagaag
780aaaacaatgt ttcattcatc tcctttaaac caagatgaaa ttaagaagcc ctcttcctag
840ttgcgggcca ggaagcgact aaccatttgt ggacaagcac agccaagtaa agccccaaat
900ctcatattgt tgtaggacaa gataaggttg acaacacacc caggttttgt ccatgttgat
960gatcaattag tttggttgtt caactgtccc gattaacatc ttcaaagcaa agcttccaac
1020agaccaacag tacattgaat gaaccgaaca aaacagaaga gaatagatgg tttctttgca
1080tttaagtttt attgtaccac ttcgactcat ttgtattcaa gttttacgaa acttcttgat
1140taactgaaaa ctgggctagg tatacttagc ccgaccaatc agatatgact tttcaccctt
1200cctttttcac caattaacta ggaaagtatc ccgcgcatgc attcgtgtgc ggggatgcat
1260atattagttt gtttcaaaaa cacatcaaaa tctaaattaa aagtaagatg ttttccttta
1320ttctaattga atgttcattg gcactctttc tttccataag caatctcgat ttttaccgct
1380gatgaatgta ttttattata atgtcaagct ggacatccct ctaataggtt taattgcata
1440tagtttacaa aattggagga aatatcatct cggtgatatt tgacaccata aaatatgatg
15001561500DNAOryza sativaOs12g42980 location Chr1226703270-26704769
156ggtttgccag aaggcagcaa cgagagagag aggcgatcga attcagtgag ctgtggcgta
60attgcccaag ccacagctct ctctctccct ctctctctct gcctataaat aagtgtttgc
120agctacgaaa attcaatcgg ggaaaaacta tatgggatta tcctattgat ttatttatgg
180tatgcaggat atggaggctt cgcaaattcg tttgttgtcg cctgtgggac atcgatatca
240gagtagaatt gtttaggctc gtactcccta tgtcaaaaaa aaaacccact tctataaatg
300aatctggaca tacacggaca tagtgtatgt ccagattcgt tttttttacg aagagagcac
360aacatatgtc tattgtcgat acttttttga gatcagtata taaaataggt ggtactagat
420taacgatggg ctagatggtt aataatgagc atatatgtgc aatagatgaa acatgtctat
480acaagtatag tggtatactg ctacaggtaa tttgttacta tgtgtcagtt caattgtaca
540gttttaaata ttattactac tatatcacaa tatgatactt gagtgttttt actatgggag
600actcccctgc tgtgttttgg ttaagagtga gcccttctca tatgagtgat tcttactctc
660ccgtcccaaa aaaaactcaa cctaggaggg gatgtgagac aacgaatctg gataaatggt
720agtccagatt cattgtacta ggaggggtca catcccctcc taagttgagt ttttttttat
780ggaggaagta tactactccc tccgtttcag gttataagac tttctaccat tgcctacatt
840catatatata tatatatata tatatatata tatgggttag aaagtcttat aatataaaac
900gaaggtagta ctgtacatat gattgggtat gggtgtttct tataattgtg actttcagat
960aataaaataa tatcccatgt tttttttaaa ctaaataata tccgatgttt ttattccatt
1020atcataagga aaaaaaatcc catgaatgat atcattgccg gcggttgcct gtggcatgca
1080agcagttggc agattcttct cgtctcacag catattgtcc cttggccctt gggcattcca
1140aatttctacc aataatttca ttctaagaac taaagtcgac gtcgccatcc cggtgcacgc
1200acgccggcca tgctagcttg cagatagaat tggaatagtt agccaagcca actccaacca
1260aagaccctac acatcaccat cctatccgtt ctacacgatg aaatattcac tccatttcta
1320atatacgttg tcatcgattt gtagtgcgtg aacagtgatt ttattaaaaa aatgtaaata
1380taaaagaata tatgtaagtc atacttaaag cagctttaat ggtaaaaata aataacaaca
1440aaaaatatta ttattattac atattttttc agtaaggtta aagataaaac atgtgtacag
15001571500DNAOryza sativaOs03g29280 location Chr316670214-16671713
(reverse complement) 157cgtggcctgc gaccgcgagt ggtcagtgtg tccttctgtg
tatagttgga atcttttcga 60tatacttctg tgtatagtta gagtttcgag gggcgagttg
aaaccttaaa agacgtaacc 120agtagaaaaa aaaacaaaca caccaccatc ataaagtaga
aaaaaaaaca aacacaccac 180catcataaac tagcaggtcc tcgcacaaca ccctaaagaa
aactctaaag tgttgtgcga 240ggacttacta gtttatgatg ataaactagt aattcctcgc
acaacaccct aaagaaaact 300ctagggtgat gtgcgagggc ctgccgtttt atgaaggtgg
tgtgtttgtt tttttttcct 360actggttacg tcttttcagg ttatgatatt atagttattt
cctgttatat ccgtacgaac 420ttcgcgctat ctaaatataa atcgtaaaaa aatatatact
ccctccgttt caaaatattt 480gacacgctat ttattaccat ggctagcaat gattttaaaa
ttgtggcaac atgtatttat 540tgctacaatt ttagtattga tgccacgatt tttttcgtgg
caacaaatga tttttctagt 600agtgattgtc aaatgtaata aataaagggt tgtgttgata
gttatcaagt ggtgttttgg 660tggcaaggtg ccagggtgct tgaggttaga ccggcgtggt
aaggcgggtc agattggaag 720tgtttgtgcg gtcagaccgg ccggatagct gtggtctaac
cataggtgtt tgagcggtca 780gactaggtag cccgacagtg ggaacgatat actcctgttt
ggagtcggta tcttgttgat 840tttatggaac atgttgattg cttaatgttt attacttcta
gtttgtttct atacgtgaga 900tatattgtac gttgtgtacc attgttgagt caagttgata
aaaaaacttg tgcttggata 960tagtttctta gtttgttcat gtgtagttgt tacttgaggc
ctcgggagta ttgccggtga 1020tgaatcgaga ctaagcttgg gaaaagttga ggtctagatg
gacaagaaga tcatgcatga 1080tactaaggct agagaacaat gcacgtagag ataaagtttc
catatggcat gcaagaagag 1140ttttgtgcgt atgagaagtg gagtcaaatt tagattggag
tccaagttta ggaagattag 1200agattgaaga tgtacaggga tgtgtatatc cttgttttga
gaagtttcct atgtaattag 1260gattccttgt tctagttgga ttcgtggcat gtttgtcggt
ataattagat gaggggtcga 1320ggctcaaagt aaggtgaagt gggcaacttt taggagagaa
aagttaggtt tctttttgag 1380atttcggttc tagttcgtga attgagaaag gaatgcttta
tattcccttt gtaagtataa 1440cttgatacaa taaagtttat ccacttttag atgccctttg
taagttaggt ttgtgttttt 15001581500DNAOryza sativaOs03g20650 location
Chr311684335-11685834 158tgcctgagct cccatccacc ccacccccaa ccccacgcgc
gcgatcacca ccacctgcga 60cacacacaac cccgagacgc accccccccc cccaaccctc
acgatcaaac aatcaaacac 120ctgacctacg ccgcagcaac aacaacgaca acaaccacaa
accacccaca caaacagagc 180ccacactgac cctcgtccct cggcggcggc gcatggcaag
aagaagcaga gaggagagtg 240gaggtgggga ggaggcaaga atttaggatt ccacaaaggg
gggtggtgcg cttgggccaa 300tgggggcaga caggaggcga tgcccctctc tccctctctc
tctctctctc tctctctcgc 360ttgggaagaa tccaaaaagc gatcgacgag gcgagaagcg
aaagacgagt gcgtgcgcct 420gtgctgtgcg tctccgcgcg cgcgcggtgc tggaaaagaa
agaaagaaag aggcgctgcc 480tttctagctc tgtgaccaag gttcgctttg cctttgcctt
tggtgggcag agggagagag 540agaggggggg ttttattcgc gggggagatg gcagctgcag
aatctgcaca agagagacgg 600gggccaatgt gacacaagag atcaggttat tcaggtactc
ccgccacatc agtctaaggc 660cccacgtaac agtcgcagcg tcactgctct ctccaccgcg
gtgatttttt ttatttaccg 720ctcacgaatc tttttgagta ctggaattcg gataacaaac
gcactcaaac gacgagtact 780atgttattcc tcccttccta catcgtataa tacaagagat
tcggataaga tgtaatattt 840ttcagtacta gaatgtgtca cctctctaaa ttctttagta
gcatgaatat gtatatacta 900tttgtccata tttatagtaa taaaaaatat gatatccggt
tctggattat tgtattttga 960gacagatgaa gtagttaaat tttaaggttt tataagaatt
tattaaaaaa ggtattgttt 1020gatagcacta tagtaaaagc gcaggaattt gagacggaat
aaagagaaaa catgaggtct 1080gtttttgtag gagatatttc tttgtattcc atatgcattc
ggagccattt ctttgtttca 1140gagaatatga agctagggat ctttttccaa ctaaatttca
attatccaaa attcctcttt 1200ttttgctgtt ctaaaaggaa cctttaattt tatagtattt
ttcatcgcaa gaaaaagtta 1260taagtgcgtt gttatatgtt ttcgagaata ccgaatttga
tgcacaacgg gttgccacag 1320aagattcagg tcctaggtgt gtcttcagtt ccgaatctaa
tacgagtggc tgcctgtact 1380actgcctccc tttcacaata taagtcattt tagcattttt
catattcata ttgatgttaa 1440tgatcaatat gaatataaaa aatatagtat tttctatatt
aatattgatg ttaatgaatt 15001591500DNAOryza sativaOs06g43920 location
Chr626453545-26455044 (reverse complement) 159tatgcaatca aagaagagat
gttgaattga ctcattagag ttacaaaagc tacaaagcaa 60gctacccttc catcttcttt
taacaagatt atcttttgtt agaatcaccc ccttttcaat 120ataccaaagg aaaattttaa
tttttaaagg aatccttagc ttccaaatta aagatttcct 180atgtattaca ccattaagca
taagggcttt atacatggag ttaacataaa acaacccttt 240tttattgctc ttgcaaaaga
attggtcttg cttatcattc aaattgacac tcaccacctt 300tgaaacaatt tcaagccaat
cttccaagtt tttccctaca attgctcttc taaaagaaac 360attcaaaggg acccttccta
acacatcggc cactaccaag tttttcccta ccattgtttt 420catctagttg atcaaaaaga
tcttgctgat caactgtgtc atcgagcaca gggagaagaa 480tgccatcttc atccccacca
ttcagcatct gagccggcat ctcagggctg tcgagatgaa 540agtcaggaaa tgtagaaccg
aagcttgaat cagttggtgt tgaagaactg atgttcgagt 600cactaccaag tgagaagttg
tcatgcatca tctctgctgg gcttgtgtcg tttcctatgt 660tgaatgaccc catctgatca
tcgaaatctg acagtgatat ggtgctcatg attggtgaca 720tttgaccatg tagttcatct
ccaaacatcg gttcctggcc actggaggct gcaaagcttg 780agagctggtt gatattggcg
atgttcgcct gcgacgggcg gaaagatgtc ccatcattgt 840gaccaagatc aggaaacatc
gatggtgctg taggttgcca gtatttgcta cagtttgcaa 900ggtccggaaa ttggtaatca
ttgccaattg tgctgaagtg gtttgtagaa gattgattca 960tctgaaccaa aggctgatta
ttcacttcag ggtactgcac tggaaactga tttgttggcg 1020ccaaaaactc accatttgac
tgcactggaa actggtttgc aggcaccgaa aacccaccat 1080ttgactgcac ttgatactga
tttgcaggcg ccgaaaaccc acaatttgac tgcactggaa 1140actgatttgc aagagctgaa
aacccaaact ttgactgtac ttgaaactga tttgcaggcg 1200ccgaaacccc attatttgac
ttgtttgcag acgagttgcc agaatggcaa gatggagagc 1260cattgcttgt ctcccacata
ctcttacaga acatgccagc atatgagcta ccagaatcgt 1320ctgtatccaa caccaagcca
tttgatatgt ttgcaaatga gctcccagat gggccagatg 1380ggaagcactt tcttgcattt
gatgagttct gcagtggctg aaacacagga ttgttgcccg 1440ggcttccacc atggccaact
gtccccatgg ccatgtttct ctggctgttc ttgagctgaa 15001606PRTArabidopsis
thalianamisc_feature(2)..(2)Xaa can be any naturally occurring amino acid
160Pro Xaa Phe Xaa Xaa Trp 1 5
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