Patent application title: Transgenic Plants with Increased Stress Tolerance and Yield
Inventors:
Amber Shirley (Durham, NC, US)
Basf Plant Science Gmbh (Ludwigshafen, DE)
Assignees:
BASF Plant Science GmbH
IPC8 Class: AC12N1582FI
USPC Class:
800290
Class name: Multicellular living organisms and unmodified parts thereof and related processes method of introducing a polynucleotide molecule into or rearrangement of genetic material within a plant or plant part the polynucleotide alters plant part growth (e.g., stem or tuber length, etc.)
Publication date: 2013-05-30
Patent application number: 20130139281
Abstract:
Polynucleotides are disclosed which are capable of enhancing a growth,
yield under water-limited conditions, and/or increased tolerance to an
environmental stress of a plant transformed to contain such
polynucleotides. Also provided are methods of using such polynucleotides
and transgenic plants and agricultural products, including seeds,
containing such polynucleotides as transgenes.Claims:
1-9. (canceled)
10. A transgenic plant transformed with an expression cassette comprising an isolated polynucleotide encoding a polypeptide comprising SEQ ID NO: 16.
11. The transgenic plant of claim 10, wherein the polynucleotide comprises SEQ ID NO:15.
12. A method of producing a transgenic plant comprising the steps of: a) introducing into a plant cell an expression vector comprising a polynucleotide encoding a polypeptide comprising SEQ ID NO: 16; and b) generating from the plant cell a transgenic plant that expresses the polynucleotide, wherein expression of the polynucleotide in the transgenic plant results in increased growth or yield of the plant under normal or water-limited conditions or increased tolerance to environmental stress as compared to a wild type variety of plant.
13. The method of claim 12, wherein the polynucleotide comprises SEQ ID NO:15.
Description:
[0001] This application claims priority benefit of U.S. provisional patent
application Ser. No. 60/953,562, filed Aug. 2, 2007, the contents of
which are hereby incorporated by reference.
FIELD OF THE INVENTION
[0002] This invention relates generally to transgenic plants which overexpress nucleic acid sequences encoding polypeptides capable of conferring increased stress tolerance and consequently, increased plant growth and crop yield, under normal or abiotic stress conditions. Additionally, the invention relates to novel isolated nucleic acid sequences encoding polypeptides that confer upon a plant increased tolerance under abiotic stress conditions, and/or increased plant growth and/or increased yield under normal or abiotic stress conditions.
BACKGROUND OF THE INVENTION
[0003] Abiotic environmental stresses, such as drought, salinity, heat, and cold, are major limiting factors of plant growth and crop yield. Crop yield is defined herein as the number of bushels of relevant agricultural product (such as grain, forage, or seed) harvested per acre. Crop losses and crop yield losses of major crops such as soybean, rice, maize (corn), cotton, and wheat caused by these stresses represent a significant economic and political factor and contribute to food shortages in many underdeveloped countries.
[0004] Water availability is an important aspect of the abiotic stresses and their effects on plant growth. Continuous exposure to drought conditions causes major alterations in the plant metabolism which ultimately lead to cell death and consequently to yield losses. Because high salt content in some soils results in less water being available for cell intake, high salt concentration has an effect on plants similar to the effect of drought on plants. Additionally, under freezing temperatures, plant cells lose water as a result of ice formation within the plant. Accordingly, crop damage from drought, heat, salinity, and cold stress, is predominantly due to dehydration.
[0005] Because plants are typically exposed to conditions of reduced water availability during their life cycle, most plants have evolved protective mechanisms against desiccation caused by abiotic stresses. However, if the severity and duration of desiccation conditions are too great, the effects on development, growth, plant size, and yield of most crop plants are profound. Developing plants efficient in water use is therefore a strategy that has the potential to significantly improve human life on a worldwide scale.
[0006] Traditional plant breeding strategies are relatively slow and require abiotic stress-tolerant founder lines for crossing with other germplasm to develop new abiotic stress-resistant lines. Limited germplasm resources for such founder lines and incompatibility in crosses between distantly related plant species represent significant problems encountered in conventional breeding. Breeding for tolerance has been largely unsuccessful.
[0007] Many agricultural biotechnology companies have attempted to identify genes that could confer tolerance to abiotic stress responses, in an effort to develop transgenic abiotic stress-tolerant crop plants. Although some genes that are involved in stress responses or water use efficiency in plants have been characterized, the characterization and cloning of plant genes that confer stress tolerance and/or water use efficiency remains largely incomplete and fragmented. To date, success at developing transgenic abiotic stress-tolerant crop plants has been limited, and no such plants have been commercialized.
[0008] In order to develop transgenic abiotic stress-tolerant crop plants, it is necessary to assay a number of parameters in model plant systems, greenhouse studies of crop plants, and in field trials. For example, water use efficiency (WUE), is a parameter often correlated with drought tolerance. Studies of a plant's response to desiccation, osmotic shock, and temperature extremes are also employed to determine the plant's tolerance or resistance to abiotic stresses. When testing for the impact of the presence of a transgene on a plant's stress tolerance, the ability to standardize soil properties, temperature, water and nutrient availability and light intensity is an intrinsic advantage of greenhouse or plant growth chamber environments compared to the field.
[0009] WUE has been defined and measured in multiple ways. One approach is to calculate the ratio of whole plant dry weight, to the weight of water consumed by the plant throughout its life. Another variation is to use a shorter lime interval when biomass accumulation and water use are measured. Yet another approach is to use measurements from restricted parts of the plant, for example, measuring only aerial growth and water use. WUE also has been defined as the ratio of CO2 uptake to water vapor loss from a leaf or portion of a leaf, often measured over a very short time period (e.g. seconds/minutes). The ratio of 13C/12C fixed in plant tissue, and measured with an isotope ratio mass-spectrometer, also has been used to estimate WUE in plants using C3 photosynthesis.
[0010] An increase in WUE is informative about the relatively improved efficiency of growth and water consumption, but this information taken alone does not indicate whether one of these two processes has changed or both have changed. In selecting traits for improving crops, an increase in WUE due to a decrease in water use, without a change in growth would have particular merit in an irrigated agricultural system where the water input costs were high. An increase in WUE driven mainly by an increase in growth without a corresponding jump in water use would have applicability to all agricultural systems. In many agricultural systems where water supply is not limiting, an increase in growth, even if it came at the expense of an increase in water use (i.e. no change in WUE), could also increase yield. Therefore, new methods to increase both WUE and biomass accumulation are required to improve agricultural productivity.
[0011] Concomitant with measurements of parameters that correlate with abiotic stress tolerance are measurements of parameters that indicate the potential impact of a transgene on crop yield. For forage crops like alfalfa, silage corn, and hay, the plant biomass correlates with the total yield. For grain crops, however, other parameters have been used to estimate yield, such as plant size, as measured by total plant dry weight, above-ground dry weight, above-ground fresh weight, leaf area, stem volume, plant height, rosette diameter, leaf length, root length, root mass, tiller number, and leaf number. Plant size at an early developmental stage will typically correlate with plant size later in development. A larger plant with a greater leaf area can typically absorb more light and carbon dioxide than a smaller plant and therefore will likely gain a greater weight during the same period. This is in addition to the potential continuation of the micro-environmental or genetic advantage that the plant had to achieve the larger size initially. There is a strong genetic component to plant size and growth rate, and so for a range of diverse genotypes plant size under one environmental condition is likely to correlate with size under another. In this way a standard environment is used to approximate the diverse and dynamic environments encountered at different locations and times by crops in the field.
[0012] Harvest index, the ratio of seed yield to above-ground dry weight, is relatively stable under many environmental conditions and so a robust correlation between plant size and grain yield is possible. Plant size and grain yield are intrinsically linked, because the majority of grain biomass is dependent on current or stored photosynthetic productivity by the leaves and stem of the plant. Therefore, selecting for plant size, even at early stages of development, has been used as to screen for for plants that may demonstrate increased yield when exposed to field testing. As with abiotic stress tolerance, measurements of plant size in early development, under standardized conditions in a growth chamber or greenhouse, are standard practices to measure potential yield advantages conferred by the presence of a transgene.
[0013] There is a need, therefore, to identify additional genes expressed in stress tolerant plants and/or plants that are efficient in water use that have the capacity to confer stress tolerance and/or increased water use efficiency to the host plant and to other plant species. Newly generated stress tolerant plants and/or plants with increased water use efficiency will have many advantages, such as an increased range in which the crop plants can be cultivated, by for example, decreasing the water requirements of a plant species. Other desirable advantages include increased resistance to lodging, the bending of shoots or stems in response to wind, rain, pests, or disease.
SUMMARY OF THE INVENTION
[0014] The present inventors have discovered that transforming a plant with certain polynucleotides results in enhancement of the plant's growth and response to environmental stress, and accordingly the yield of the agricultural products of the plant is increased, when the polynucleotides are present in the plant as transgenes. The polynucleotides capable of mediating such enhancements have been isolated from Brassica napus, Oryza sativa, Glycine max, Triticum aestivum, Hordeum vulgare, Zea mays, and Linum usitatissimum and are listed in Table 1, and the sequences thereof are set forth in the Sequence Listing as indicated in Table 1.
TABLE-US-00001 TABLE 1 Polynucleotide Amino acid Gene ID Organism SEQ ID NO SEQ ID NO BN51364980 B. napus 1 2 OS34096188 O. sativa 3 4 OS32583643 O. sativa 5 6 GM53626178 G. max 7 8 TA56540264 T. aestivum 9 10 BN45206322 B. napus 11 12 GM48923793 G. max 13 14 TA55969932 T. aestivum 15 16 BN47310186 B. napus 17 18 BN51359456 B. napus 19 20 HV62552639 H. vulgare 21 22 ZM61995511 Z. mays 23 24 LU61567101 L. usitatissimum 25 26 LU61893412 L. usitatissimum 27 28 OS39781852 O. sativa 29 30 OS34701560 O. sativa 31 32 OS36821256 O. sativa 33 34 GM51659494 G. max 35 36 GM49780101 G. max 37 38 GM59637305 G. max 39 40 TA55974113 T. aestivum 41 42
[0015] In one embodiment, the invention provides a transgenic plant transformed with an expression cassette comprising an isolated polynucleotide encoding a methionine sulfoxide reductase family protein selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, and SEQ ID NO:10, or a SelR protein domain.
[0016] In another embodiment, the invention provides a transgenic plant transformed with an expression cassette comprising an isolated polynucleotide encoding a homeodomain leucine zipper protein having a sequence as set forth in SEQ ID NO:12, SEQ ID NO:14, or SEQ ID NO: 16.
[0017] In another embodiment, the invention provides a transgenic plant transformed with an expression cassette comprising an isolated polynucleotide encoding a zinc finger protein containing an A20 domain in combination with an AN1 domain selected from the group consisting of SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, and SEQ ID NO:44, or both AN1-like and A20-like zinc finger protein domains.
[0018] In a further embodiment, the invention concerns a seed produced by the transgenic plant of the invention, wherein the seed is true breeding for a transgene comprising the polynucleotide described above. Plants derived from the seed of the invention demonstrate increased tolerance to an environmental stress, and/or increased plant growth, and/or increased yield, under normal or stress conditions as compared to a wild type variety of the plant.
[0019] In a still another aspect, the invention concerns products produced by or from the transgenic plants of the invention, their plant parts, or their seeds, such as a foodstuff, feedstuff, food supplement, feed supplement, cosmetic or pharmaceutical.
[0020] The invention further provides the isolated polynucleotides identified in Table 1 below, and polypeptides identified in Table 1. The invention is also embodied in recombinant vector comprising an isolated polynucleotide of the invention.
[0021] In yet another embodiment, the invention concerns a method of producing the aforesaid transgenic plant, wherein the method comprises transforming a plant cell with an expression vector comprising an isolated polynucleotide of the invention, and generating from the plant cell a transgenic plant that expresses the polypeptide encoded by the polynucleotide. Expression of the polypeptide in the plant results in increased tolerance to an environmental stress, and/or growth, and/or yield under normal and/or stress conditions as compared to a wild type variety of the plant.
[0022] In still another embodiment, the invention provides a method of increasing a plant's tolerance to an environmental stress, and/or growth, and/or yield. The method comprises the steps of transforming a plant cell with an expression cassette comprising an isolated polynucleotide of the invention, and generating a transgenic plant from the plant cell, wherein the transgenic plant comprises the polynucleotide.
BRIEF DESCRIPTION OF THE DRAWINGS
[0023] FIG. 1 is an alignment of BN51364980, OS34096188, OS32583643, GM53626178, TA56540264 with a known methionine sulfoxide reductase family protein.
[0024] FIG. 2 is an alignment of BN45206322, GM48923793, and TA55969932 with a known homeodomain leucine zipper protein.
[0025] FIG. 3 is an alignment of BN47310186, BN51359456, HV62552639, ZM61995511, LU61567101, LU61893412, OS39781852, OS34701560, OS36821256, GM51659494, GM49780101, GM59637305, and TA55974113 with a known A20 and AN1 domain containing zinc finger protein.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0026] Throughout this application, various publications are referenced. The disclosures of all of these publications and those references cited within those publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains. The terminology used herein is for the purpose of describing specific embodiments only and is not intended to be limiting. As used herein, "a" or "an" can mean one or more, depending upon the context in which it is used. Thus, for example, reference to "a cell" can mean that at least one cell can be used.
[0027] In one embodiment, the invention provides a transgenic plant that overexpresses an isolated polynucleotide identified in Table 1, or a homolog thereof. The transgenic plant of the invention demonstrates an increased tolerance to an environmental stress as compared to a wild type variety of the plant. The overexpression of such isolated nucleic acids in the plant may optionally result in an increase in plant growth or in yield of associated agricultural products, under normal or stress conditions, as compared to a wild type variety of the plant. Without wishing to be bound by any theory, the increased tolerance to an environmental stress, increased growth, and/or increased yield of a transgenic plant of the invention is believed to result from an increase in water use efficiency of the plant.
[0028] As defined herein, a "transgenic plant" is a plant that has been altered using recombinant DNA technology to contain an isolated nucleic acid which would otherwise not be present in the plant. As used herein, the term "plant" includes a whole plant, plant cells, and plant parts. Plant parts include, but are not limited to, stems, roots, ovules, stamens, leaves, embryos, meristematic regions, callus tissue, gametophytes, sporophytes, pollen, microspores, and the like. The transgenic plant of the invention may be male sterile or male fertile, and may further include transgenes other than those that comprise the isolated polynucleotides described herein.
[0029] As used herein, the term "variety" refers to a group of plants within a species that share constant characteristics that separate them from the typical form and from other possible varieties within that species. While possessing at least one distinctive trait, a variety is also characterized by some variation between individuals within the variety, based primarily on the Mendelian segregation of traits among the progeny of succeeding generations. A variety is considered "true breeding" for a particular trait if it is genetically homozygous for that trait to the extent that, when the true-breeding variety is self-pollinated, a significant amount of independent segregation of the trait among the progeny is not observed. In the present invention, the trait arises from the transgenic expression of one or more isolated polynucleotides introduced into a plant variety. As also used herein, the term "wild type variety" refers to a group of plants that are analyzed for comparative purposes as a control plant, wherein the wild type variety plant is identical to the transgenic plant (plant transformed with an isolated polynucleotide in accordance with the invention) with the exception that the wild type variety plant has not been transformed with an isolated polynucleotide of the invention.
[0030] As defined herein, the term "nucleic acid" and "polynucleotide" are interchangeable and refer to RNA or DNA that is linear or branched, single or double stranded, or a hybrid thereof. The term also encompasses RNA/DNA hybrids. An "isolated" nucleic acid molecule is one that is substantially separated from other nucleic acid molecules which are present in the natural source of the nucleic acid (i.e., sequences encoding other polypeptides). For example, a cloned nucleic acid is considered isolated. A nucleic acid is also considered isolated if it has been altered by human intervention, or placed in a locus or location that is not its natural site, or if it is introduced into a cell by transformation. Moreover, an isolated nucleic acid molecule, such as a cDNA molecule, can be free from some of the other cellular material with which it is naturally associated, or culture medium when produced by recombinant techniques, or chemical precursors or other chemicals when chemically synthesized. While it may optionally encompass untranslated sequence located at both the 3' and 5' ends of the coding region of a gene, it may be preferable to remove the sequences which naturally flank the coding region in its naturally occurring replicon.
[0031] As used herein, the term "environmental stress" refers to a sub-optimal condition associated with salinity, drought, nitrogen, temperature, metal, chemical, pathogenic, or oxidative stresses, or any combination thereof. The terms "water use efficiency" and "WUE" refer to the amount of organic matter produced by a plant divided by the amount of water used by the plant in producing it, i.e., the dry weight of a plant in relation to the plant's water use. As used herein, the term "dry weight" refers to everything in the plant other than water, and includes, for example, carbohydrates, proteins, oils, and mineral nutrients.
[0032] Any plant species may be transformed to create a transgenic plant in accordance with the invention. The transgenic plant of the invention may be a dicotyledonous plant or a monocotyledonous plant. For example and without limitation, transgenic plants of the invention may be derived from any of the following diclotyledonous plant families: Leguminosae, including plants such as pea, alfalfa and soybean; Umbelliferae, including plants such as carrot and celery; Solanaceae, including the plants such as tomato, potato, aubergine, tobacco, and pepper; Cruciferae, particularly the genus Brassica, which includes plant such as oilseed rape, beet, cabbage, cauliflower and broccoli); and Arabidopsis thaliana; Compositae, which includes plants such as lettuce; Malvaceae, which includes cotton; Fabaceae, which includes plants such as peanut, and the like. Transgenic plants of the invention may be derived from monocotyledonous plants, such as, for example, wheat, barley, sorghum, millet, rye, triticale, maize, rice, oats and sugarcane. Transgenic plants of the invention are also embodied as trees such as apple, pear, quince, plum, cherry, peach, nectarine, apricot, papaya, mango, and other woody species including coniferous and deciduous trees such as poplar, pine, sequoia, cedar, oak, and the like. Especially preferred are Arabidopsis thaliana, Nicotiana tabacum, oilseed rape, soybean, corn (maize), wheat, linseed, potato and tagetes.
[0033] As shown in Table 1, one embodiment of the invention is a transgenic plant transformed with an expression cassette comprising an isolated polynucleotide encoding a methionine sulfoxide reductase family protein. Methionine sulfoxide reductases (MSRs) catalyze the thioredoxin-dependent reduction of methionine sulfoxide (MetSO) to the correct methionine residue. Methionine is highly susceptible to oxidative damage, and methionine oxidation results in modification of the activity and conformation of many proteins.
[0034] There are two types of MSRs, type A and type B; however, these two types are unrelated in both sequence and structure. The MSRB enzyme selectively catalytically reduces the MetSO R enantiomer. MSRB type proteins contain four conserved cysteine residues in two CxxC motifs, where x can be any amino acid. These CxxC motifs are potentially involved in zinc fixation.
[0035] There are described examples from several plant species where environmental stress conditions result in increased reactive oxygen species (ROS) levels and resulting oxidative damage leads to modification in MSR gene expression. MSRs themselves are good candidates for direct antioxidants since cyclic oxidation and reduction of methionine residues could function as an efficient pathway to remove ROS in cells. in eukaryotes, senescence and a host of diseases are triggered by methionine oxidation resulting in the disruption of protein structure and function. The substrates of MSR proteins are largely unknown. To date, the first plant MSR substrate to have been identified is the small, plastidic heat shock protein Hsp21. Hsp21 contains a conserved N-terminal region that is highly enriched in methionine residues. This methinine region must be maintained in the reduced form in order to maintain the chaperone-like activity of Hsp21.
[0036] The transgenic plant of this embodiment may comprise any polynucleotide encoding a methionine sulfoxide reductase family protein. Preferably, the transgenic plant of this embodiment comprises a polynucleotide encoding a SelR domain having a sequence comprising amino acids 77 to 199 of SEQ ID NO:2; amino acids 79 to 200 of SEQ ID NO: 4; amino acids 91 to 213 of SEQ ID NO: 6; amino acids 79 to 200 of SEQ ID NO: 8; amino acids 80 to 201 of SEQ ID NO: 10. More preferably, the transgenic plant of this embodiment comprises a polynucleotide encoding a methionine sulfoxide reductase family protein having a sequence comprising amino acids 1 to 205 of SEQ ID NO:2; amino acids 1 to 204 of SEQ ID NO: 4; amino acids 1 to 214 of SEQ ID NO: 6; amino acids 1 to 202 of SEQ ID NO: 8; amino acids 1 to 206 of SEQ ID NO: 10.
[0037] In another embodiment, the invention provides a transgenic plant transformed with an expression cassette comprising an isolated polynucleotide encoding a homeodomain leucine zipper protein. Homeodomain leucine zipper (HDZip) proteins belong to a family of transcription factors that interact as dimers via a leucine zipper domain and bind DNA in a sequence specific manner via their homeodomains. Based upon sequence, the HDZip family proteins are divided into four classes. The Class I HDZip proteins are suggested to regulate plant response to ABA and may have regulatory roles related to ABA signalling. The Class I HDZip protein members can form heterodimers in vitro; therefore, this class may constitute an interacting network of proteins that mediates responses to environmental stimuli and/or integrates signals to regulate similar sets of target genes.
[0038] The transgenic plant of this embodiment may comprise any polynucleotide encoding a homeodomain leucine zipper protein. Preferably, the transgenic plant of this embodiment comprises a polynucleotide encoding a homeobox domain having a sequence comprising amino acids 62 to 116 of SEQ ID NO:12; amino acids 83 to 137 of SEQ ID NO: 14; amino acids 76 to 130 of SEQ ID NO: 16 or a homeobox associated leucine zipper domain having a sequence comprising amino acids 117 to 161 of SEQ ID NO: 12; amino acids 138 to 182 of SEQ ID NO: 14; amino acids 131 to 175 of SEQ ID NO: 16. More preferably, the transgenic plant of this embodiment comprises a polynucleotide encoding a homeodomain leucine zipper protein having a sequence comprising amino acids 1 to 310 of SEQ ID NO:12; amino acids 1 to 331 of SEQ ID NO: 14; amino acids 1 to 340 of SEQ ID NO: 16.
[0039] In another embodiment, the invention provides a transgenic plant transformed with an expression cassette comprising an isolated polynucleotide encoding a A20 and AN1 domain containing zinc finger protein. The A20 and AN1 domain containing zinc finger proteins are found in all eukaryotes. These proteins are characterized by the presence of an A20 zinc finger domain containing multiple Cysteine2/Cysteine2 finger motifs and an AN1 zinc finger domain. The AN1 domain is usually found in proteins containing the A20 zinc finger domain. The function of these proteins is well characterized in animal systems, but little is known about the function of these proteins in plants.
[0040] The rice OsiSAP1 protein was identified as the first plant protein having both A20 and AN1 zinc finger domains. This protein was found to be associated with multiple stresses. The OsiSAP1 gene is induced in response to environmental stresses such as cold, salt, drought, submergence, wounding, and heavy metals. An ortholog from bean is also known to be inducible when eliciter treated and in response to wounding. When overexpressed in tobacco, OsiSAP1 engenders abiotic stress tolerance. OsiSAP1 does not have a typical nuclear localization signal and thus is believed to function via the zinc finger domains for protein-protein interaction.
[0041] The transgenic plant of this embodiment may comprise any polynucleotide encoding a zinc finger protein containing an A20 domain in combination with an AN1 domain. Preferably, the transgenic plant of this embodiment comprises a polynucleotide encoding an A20-like zinc finger having a sequence comprising amino acids 15 to 39 of SEQ ID NO:18; amino acids 13 to 37 of SEQ ID NO: 20; amino acids 15 to 39 of SEQ ID NO: 22; amino acids 14 to 38 of SEQ ID NO: 24; amino acids 14 to 38 of SEQ ID NO: 26; amino acids 40 to 64 of SEQ ID NO: 28; amino acids 15 to 39 of SEQ ID NO: 30; amino acids 19 to 43 of SEQ ID NO: 32; amino acids 13 to 37 of SEQ ID NO: 34; amino acids 19 to 43 of SEQ ID NO: 36; amino acids 18 to 42 of SEQ ID NO: 38; amino acids 15 to 39 of SEQ ID NO: 40; amino acids 15 to 39 of SEQ ID NO: 42; amino acids 19 to 43 of SEQ ID NO: 44 and an AN1-like zinc finger domain having a sequence comprising amino acids 118 to 158 of SEQ ID NO: 18; amino acids 128 to 168 of SEQ ID NO: 20; amino acids 95 to 135 of SEQ ID NO: 22; amino acids 112 to 152 of SEQ ID NO: 24; amino acids 115 to 155 of SEQ ID NO: 26; amino acids 179 to 219 of SEQ ID NO: 28; amino acids 110 to 150 of SEQ ID NO: 30; amino acids 105 to 145 of SEQ ID NO: 32; amino acids 105 to 145 of SEQ ID NO: 34; amino acids 111 to 151 of SEQ ID NO: 36; amino acids 102 to 142 of SEQ ID NO: 38; amino acids 113 to 153 of SEQ ID NO: 40; amino acids 115 to 155 of SEQ ID NO: 42; amino acids 106 to 146 of SEQ ID NO: 44. More preferably, the transgenic plant of this embodiment comprises a polynucleotide encoding a A20 and AN1 domain containing zinc finger protein having a sequence comprising amino acids 1 to 177 of SEQ ID NO:18; amino acids 1 to 187 of SEQ ID NO: 20; amino acids 1 to 154 of SEQ ID NO: 22; amino acids 1 to 171 of SEQ ID NO: 24; amino acids 1 to 174 of SEQ ID NO: 26; amino acids 1 to 239 of SEQ ID NO: 28; amino acids 1 to 169 of SEQ ID NO: 30; amino acids 1 to 164 of SEQ ID NO: 32; amino acids 1 to 164 of SEQ ID NO: 34; amino acids 1 to 170 of SEQ ID NO: 36; amino acids 1 to 161 of SEQ ID NO: 38; amino acids 1 to 172 of SEQ ID NO: 40; amino acids 1 to 174 of SEQ ID NO: 42, amino acids 1 to 165 of SEQ ID NO: 44.
[0042] The invention further provides a seed produced by a transgenic plant expressing polynucleotide listed in Table 1, wherein the seed contains the polynucleotide, and wherein the plant is true breeding for increased growth and/or yield under normal or stress conditions and/or increased tolerance to an environmental stress as compared to a wild type variety of the plant. The invention also provides a product produced by or from the transgenic plants expressing the polynucleotide, their plant parts, or their seeds. The product can be obtained using various methods well known in the art. As used herein, the word "product" includes, but not limited to, a foodstuff, feedstuff, a food supplement, feed supplement, cosmetic or pharmaceutical. Foodstuffs are regarded as compositions used for nutrition or for supplementing nutrition. Animal feedstuffs and animal feed supplements, in particular, are regarded as foodstuffs. The invention further provides an agricultural product produced by any of the transgenic plants, plant parts, and plant seeds. Agricultural products include, but are not limited to, plant extracts, proteins, amino acids, carbohydrates, fats, oils, polymers, vitamins, and the like.
[0043] In a preferred embodiment, an isolated polynucleotide of the invention comprises a polynucleotide having a sequence selected from the group consisting of the polynucleotide sequences listed in Table 1. These polynucleotides may comprise sequences of the coding region, as well as 5' untranslated sequences and 3' untranslated sequences.
[0044] A polynucleotide of the invention can be isolated using standard molecular biology techniques and the sequence information provided herein. Synthetic oligonucleotide primers for polymerase chain reaction amplification can be designed based upon the nucleotide sequence shown in Table 1. A nucleic acid molecule of the invention can be amplified using cDNA or, alternatively, genomic DNA, as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques. The nucleic acid molecule so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis. Furthermore, oligonucleotides corresponding to the nucleotide sequences listed in Table 1 can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer.
[0045] "Homologs" are defined herein as two nucleic acids or polypeptides that have similar, or substantially identical, nucleotide or amino acid sequences, respectively. Homologs include allelic variants, analogs, and orthologs, as defined below. As used herein, the term "analogs" refers to two nucleic acids that have the same or similar function, but that have evolved separately in unrelated organisms. As used herein, the term "orthologs" refers to two nucleic acids from different species, but that have evolved from a common ancestral gene by speciation. The term homolog further encompasses nucleic acid molecules that differ from one of the nucleotide sequences shown in Table 1 due to degeneracy of the genetic code and thus encode the same polypeptide. As used herein, a "naturally occurring" nucleic acid molecule refers to an RNA or DNA molecule having a nucleotide sequence that occurs in nature (e.g., encodes a natural polypeptide).
[0046] To determine the percent sequence identity of two amino acid sequences (e.g., one of the polypeptide sequences of Table 1 and a homolog thereof), the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of one polypeptide for optimal alignment with the other polypeptide or nucleic acid). The amino acid residues at corresponding amino acid positions are then compared. When a position in one sequence is occupied by the same amino acid residue as the corresponding position in the other sequence then the molecules are identical at that position. The same type of comparison can be made between two nucleic acid sequences.
[0047] Preferably, the isolated amino acid homologs, analogs, and orthologs of the polypeptides of the present invention are at least about 50-60%, preferably at least about 6070%, and more preferably at least about 70-75%, 75-80%, 80-85%, 85-90%, or 90-95%, and most preferably at least about 96%, 97%, 98%, 99%, or more identical to an entire amino acid sequence identified in Table 1. In another preferred embodiment, an isolated nucleic acid homolog of the invention comprises a nucleotide sequence which is at least about 40-60%, preferably at least about 60-70%, more preferably at least about 70-75%, 75-80%, 80-85%, 85-90%, or 90-95%, and even more preferably at least about 95%, 96%, 97%, 98%, 99%, or more identical to a nucleotide sequence shown in Table 1.
[0048] The percent sequence identity between two nucleic acid or polypeptide sequences may be determined using, for example, the Vector NTI 9.0 (PC) software package (Invitrogen, 1600 Faraday Ave., Carlsbad, Calif. 92008). When Vector NTI is used, a gap opening penalty of 15 and a gap extension penalty of 6.66 may be used for determining the percent identity of two nucleic acids and a gap opening penalty of 10 and a gap extension penalty of 0.1 may be used for determining the percent identity of two polypeptides. All other Vector NTI parameters may be set at the default settings. For purposes of a multiple alignment (Clustal W algorithm) using Vector NTI, the gap opening penalty is 10, and the gap extension penalty is 0.05 with blosum62 matrix. Alternatively, Align 2.0 (Myers and Miller (1989) CABIOS 4, 11-17) may be used, with all parameters set to default settings. It is to be understood that for the purposes of determining sequence identity when comparing a DNA sequence to an RNA sequence, a thymidine nucleotide is equivalent to a uracil nucleotide.
[0049] Nucleic acid molecules corresponding to homologs, analogs, and orthologs of the polypeptides listed in Table 1 can be isolated based on their identity to said polypeptides, using the polynucleotides encoding the respective polypeptides or primers based thereon, as hybridization probes according to standard hybridization techniques under stringent hybridization conditions. As used herein with regard to hybridization for DNA to a DNA blot, the term "stringent conditions" refers to hybridization overnight at 60° C. in 10×Denhart's solution, 6×SSC, 0.5% SDS, and 100 μg/ml denatured salmon sperm DNA. Blots are washed sequentially at 62° C. for 30 minutes each time in 3×SSC/0.1% SDS, followed by 1×SSC/0.1% SDS, and finally 0.1×SSC/0.1% SDS. As also used herein, in a preferred embodiment, the phrase "stringent conditions" refers to hybridization in a 6×SSC solution at 65° C. In another embodiment, "highly stringent conditions" refers to hybridization overnight at 65° C. in 10×Denhart's solution, 6×SSC, 0.5% SDS and 100 μg/ml denatured salmon sperm DNA. Blots are washed sequentially at 65° C. for 30 minutes each time in 3×SSC/0.1% SDS, followed by 1×SSC/0.1% SDS, and finally 0.1×SSC/0.1% SDS. Methods for nucleic acid hybridizations are described in Meinkoth and Wahl, 1984, Anal. Biochem. 138:267-284; well known in the art (see, for example, Current Protocols in Molecular Biology, Chapter 2, Ausubel et al., eds., Greene Publishing and Wiley-Interscience, New York, 1995; and Tijssen, 1993, Laboratory Techniques in Biochemistry and Molecular Biology: Hybridization with Nucleic Acid Probes, Part I, Chapter 2, Elsevier, New York, 1993). Preferably, an isolated nucleic acid molecule of the invention that hybridizes under stringent or highly stringent conditions to a nucleotide sequence listed in Table 1 corresponds to a naturally occurring nucleic acid molecule.
[0050] There are a variety of methods that can be used to produce libraries of potential homologs from a degenerate oligonucleotide sequence. Chemical synthesis of a degenerate gene sequence can be performed in an automatic DNA synthesizer, and the synthetic gene is then ligated into an appropriate expression vector. Use of a degenerate set of genes allows for the provision, in one mixture, of all of the sequences encoding the desired set of potential sequences. Methods for synthesizing degenerate oligonucleotides are known in the art (See, e.g., Narang, 1983, Tetrahedron 39:3; Itakura et al., 1984, Annu. Rev. Biochem. 53:323; Itakura et al., 1984, Science 198:1056; Ike et al., 1983, Nucleic Acid Res. 11:477).
[0051] Additionally, optimized nucleic acids can be created. Preferably, an optimized nucleic acid encodes a polypeptide that has a function similar to those of the polypeptides listed in Table 1 and/or modulates a plant's growth and/or yield under normal and/or water-limited conditions and/or tolerance to an environmental stress, and more preferably increases a plant's growth and/or yield under normal and/or water-limited conditions and/or tolerance to an environmental stress upon its overexpression in the plant. As used herein, "optimized" refers to a nucleic acid that is genetically engineered to increase its expression in a given plant or animal. To provide plant optimized nucleic acids, the DNA sequence of the gene can be modified to: 1) comprise codons preferred by highly expressed plant genes; 2) comprise an A+T content in nucleotide base composition to that substantially found in plants; 3) form a plant initiation sequence; 4) to eliminate sequences that cause destabilization, inappropriate polyadenylation, degradation and termination of RNA, or that form secondary structure hairpins or RNA splice sites; or 5) elimination of antisense open reading frames. Increased expression of nucleic acids in plants can be achieved by utilizing the distribution frequency of codon usage in plants in general or in a particular plant. Methods for optimizing nucleic acid expression in plants can be found in EPA 0359472; EPA 0385962; PCT Application No. WO 91/16432; U.S. Pat. No. 5,380,831; U.S. Pat. No. 5,436,391; Perlack et al., 1991, Proc. Natl. Acad. Sci. USA 88:3324-3328; and Murray et al., 1989, Nucleic Acids Res. 17:477-498.
[0052] An isolated polynucleotide of the invention can be optimized such that its distribution frequency of codon usage deviates, preferably, no more than 25% from that of highly expressed plant genes and, more preferably, no more than about 10%. In addition, consideration is given to the percentage G+C content of the degenerate third base (monocotyledons appear to favor G+C in this position, whereas dicotyledons do not). It is also recognized that the XCG (where X is A, T, C, or G) nucleotide is the least preferred codon in dicots, whereas the XTA codon is avoided in both monocots and dicots. Optimized nucleic acids of this invention also preferably have CG and TA doublet avoidance indices closely approximating those of the chosen host plant. More preferably, these indices deviate from that of the host by no more than about 10-15%.
[0053] The invention further provides an isolated recombinant expression vector comprising a polynucleotide as described above, wherein expression of the vector in a host cell results in the plant's increased growth and/or yield under normal or water-limited conditions and/or increased tolerance to environmental stress as compared to a wild type variety of the host cell. The recombinant expression vectors of the invention comprise a nucleic acid of the invention in a form suitable for expression of the nucleic acid in a host cell, which means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis of the host cells to be used for expression, which is operatively linked to the nucleic acid sequence to be expressed. As used herein with respect to a recombinant expression vector, "operatively linked" is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner which allows for expression of the nucleotide sequence (e.g., in a bacterial or plant host cell when the vector is introduced into the host cell). The term "regulatory sequence" is intended to include promoters, enhancers, and other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are well known in the art. Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cells and those that direct expression of the nucleotide sequence only in certain host cells or under certain conditions. It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of polypeptide desired, etc. The expression vectors of the invention can be introduced into host cells to thereby produce polypeptides encoded by nucleic acids as described herein.
[0054] Plant gene expression should be operatively linked to an appropriate promoter conferring gene expression in a timely, cell specific, or tissue specific manner. Promoters useful in the expression cassettes of the invention include any promoter that is capable of initiating transcription in a plant cell. Such promoters include, but are not limited to, those that can be obtained from plants, plant viruses, and bacteria that contain genes that are expressed in plants, such as Agrobacterium and Rhizobium.
[0055] The promoter may be constitutive, inducible, developmental stage-preferred, cell type-preferred, tissue-preferred, or organ-preferred. Constitutive promoters are active under most conditions. Examples of constitutive promoters include the CaMV 19S and 35S promoters (Odell et al., 1985, Nature 313:810-812), the sX CaMV 35S promoter (Kay et al, 1987, Science 236:1299-1302) the Sep1 promoter, the rice actin promoter (McElroy et al., 1990, Plant Cell 2:163-171), the Arabidopsis actin promoter, the ubiquitan promoter (Christensen et al., 1989, Plant Molec. Biol. 18:675-689), pEmu (Last et al., 1991, Theor. Appl. Genet. 81:581-588), the figwort mosaic virus 35S promoter, the Smas promoter (Velten et al., 1984, EMBO J 3:2723-2730), the super promoter (U.S. Pat. No. 5, 955,646), the GRP1-8 promoter, the cinnamyl alcohol dehydrogenase promoter (U.S. Pat. No. 5,683,439), promoters from the T-DNA of Agrobacterium, such as mannopine synthase, nopaline synthase, and octopine synthase, the small subunit of ribulose biphosphate carboxylase (ssuRUBISCO) promoter, and the like.
[0056] Inducible promoters are preferentially active under certain environmental conditions, such as the presence or absence of a nutrient or metabolite, heat or cold, light, pathogen attack, anaerobic conditions, and the like. For example, the hsp80 promoter from Brassica is induced by heat shock; the PPDK promoter is induced by light; the PR-1 promoters from tobacco, Arabidopsis, and maize are inducible by infection with a pathogen; and the Adh1 promoter is induced by hypoxia and cold stress. Plant gene expression can also be facilitated via an inducible promoter (For a review, see Getz, 1997, Annu. Rev. Plant Physiol. Plant Mol. Biol. 48:89-108). Chemically inducible promoters are especially suitable if gene expression is wanted to occur in a time specific manner. Examples of such promoters are a salicylic acid inducible promoter (PCT Application No. WO 95/19443), a tetracycline inducible promoter (Getz et al., 1992, Plant J. 2: 397-404), and an ethanol inducible promoter (PCT Application No. WO 93/21334).
[0057] In one preferred embodiment of the present invention, the inducible promoter is a stress-inducible promoter. For the purposes of the invention, stress-inducible promoters are preferentially active under one or more of the following stresses: sub-optimal conditions associated with salinity, drought, nitrogen, temperature, metal, chemical, pathogenic, and oxidative stresses. Stress inducible promoters include, but are not limited to, Cor78 (Chak et al., 2000, Planta 210:875-883; Hovath et al., 1993, Plant Physiol. 103:1047-1053), Cor15a (Artus et al., 1996, PNAS 93(23):13404-09), Rci2A (Medina et al., 2001, Plant Physiol. 125:1655-66; Nylander et al., 2001, Plant Mol. Biol. 45:341-52; Navarre and Goffeau, 2000, EMBO J. 19:2515-24; Capel et al., 1997, Plant Physiol. 115:569-76), Rd22 (Xiong et al., 2001, Plant Cell 13:2063-83; Abe et al., 1997, Plant Cell 9:1859-68; Iwasaki et al., 1995, Mol. Gen. Genet. 247:391-8), cDet6 (Lang and Palve, 1992, Plant Mol. Biol. 20:951-62), ADH1 (Hoeren et al., 1998, Genetics 149:479-90), KAT1 (Nakamura et al., 1995, Plant Physiol. 109:371-4), KST1 (Muller-Rober et al., 1995, EMBO 14:2409-16), Rha1 (Terryn et al., 1993, Plant Cell 5:1761-9; Terryn et al., 1992, FEBS Lett. 299(3):287-90), ARSK1 (Atkinson et al., 1997, GenBank Accession # L22302, and PCT Application No. WO 97/20057), PtxA (Plesch et al., GenBank Accession # X67427), SbHRGP3 (Ahn et al., 1996, Plant Cell 8:1477-90), GH3 (Liu et al., 1994, Plant Cell 6:645-57), the pathogen inducible PRP1-gene promoter (Ward et al., 1993, Plant. Mol. Biol. 22:361-366), the heat inducible hsp80-promoter from tomato (U.S. Pat. No. 5,187,267), cold inducible alpha-amylase promoter from potato (PCT Application No. WO 96/12814), or the wound-inducible pinII-promoter (European Patent No. 375091). For other examples of drought, cold, and salt-inducible promoters, such as the RD29A promoter, see Yamaguchi-Shinozalei et al., 1993, Mol. Gen, Genet. 236:331-340.
[0058] Developmental stage-preferred promoters are preferentially expressed at certain stages of development. Tissue and organ preferred promoters include those that are preferentially expressed in certain tissues or organs, such as leaves, roots, seeds, or xylem. Examples of tissue-preferred and organ-preferred promoters include, but are not limited to fruit-preferred, ovule-preferred, male tissue-preferred, seed-preferred, integument-preferred, tuber-preferred, stalk-preferred, pericarp-preferred, leaf-preferred, stigma-preferred, pollen-preferred, anther-preferred, petal-preferred, sepal-preferred, pedicel-preferred, silique-preferred, stem-preferred, root-preferred promoters, and the like. Seed-preferred promoters are preferentially expressed during seed development and/or germination. For example, seed-preferred promoters can be embryo-preferred, endosperm-preferred, and seed coat-preferred (See Thompson et al., 1989, BioEssays 10:108). Examples of seed-preferred promoters include, but are not limited to, cellulose synthase (celA), Cim1, gamma-zein, globulin-1, maize 19 kD zein (cZ19B1), and the like.
[0059] Other suitable tissue-preferred or organ-preferred promoters include the napingene promoter from rapeseed (U.S. Pat. No. 5,608,152), the USP-promoter from Vicia faba (Baeumlein et al., 1991, Mol. Gen. Genet. 225(3): 459-67), the oleosin-promoter from Arabidopsis (PCT Application No. WO 98145461), the phaseolin-promoter from Phaseolus vulgaris (U.S. Pat. No. 5,504,200), the Bce4-promoter from Brassica (PCT Application No. WO 91/13980), or the legumin B4 promoter (LeB4; Baeumlein et al., 1992, Plant Journal, 2(2): 233-9), as well as promoters conferring seed specific expression in monocot plants like maize, barley, wheat, rye, rice, etc. Suitable promoters to note are the Ipt2 or Ipt1-gene promoter from barley (PCT Application No. WO 95/15389 and PCT Application No. WO 95/23230) or those described in PCT Application No. WO 99/16890 (promoters from the barley hordein-gene, rice glutelin gene, rice oryzin gene, rice prolamin gene, wheat gliadin gene, wheat glutelin gene, oat glutelin gene, Sorghum kasirin-gene, and rye secalin gene).
[0060] Other promoters useful in the expression cassettes of the invention include, but are not limited to, the major chlorophyll a/b binding protein promoter, histone promoters, the Ap3 promoter, the β-conglycin promoter, the napin promoter, the soybean lectin promoter, the maize 15 kD zein promoter, the 22 kD zein promoter, the 27 kD zein promoter, the g-zein promoter, the waxy, shrunken 1, shrunken 2, and bronze promoters, the Zm13 promoter (U.S. Pat. No. 5,086,169), the maize polygalacturonase promoters (PG) (U.S. Pat. Nos. 5,412,085 and 5,545,546), and the SGB6 promoter (U.S. Pat. No. 5,470,359), as well as synthetic or other natural promoters.
[0061] Additional flexibility in controlling heterologous gene expression in plants may be obtained by using DNA binding domains and response elements from heterologous sources (i.e., DNA binding domains from non-plant sources). An example of such a heterologous DNA binding domain is the LexA DNA binding domain (Brent and Ptashne, 1985, Cell 43:729-736).
[0062] In a preferred embodiment of the present invention, the polynucleotides listed in Table 1 are expressed in plant cells from higher plants (e.g., the spermatophytes, such as crop plants). A polynucleotide may be "introduced" into a plant cell by any means, including transfection, transformation or transduction, electroporation, particle bombardment, agroinfection, and the like. Suitable methods for transforming or transfecting plant cells are disclosed, for example, using particle bombardment as set forth in U.S. Pat. Nos. 4,945,050; 5,036,006; 5,100,792; 5,302,523; 5,464,765; 5,120,657; 6,084,154; and the like. More preferably, the transgenic corn seed of the invention may be made using Agrobacterium transformation, as described in U.S. Pat. Nos. 5,591,616; 5,731,179; 5,981,840; 5,990,387; 6,162,965; 6,420,630, U.S. patent application publication number 2002/0104132, and the like. Transformation of soybean can be performed using for example a technique described in European Patent No. EP 0424047, U.S. Pat. No. 5,322,783, European Patent No. EP 0397 687, U.S. Pat. No. 5,376,543, or U.S. Pat. No. 5,169,770. A specific example of wheat transformation can be found in PCT Application No. WO 93/07256. Cotton may be transformed using methods disclosed in U.S. Pat. Nos. 5,004,863; 5,159,135; 5,846,797, and the like. Rice may be transformed using methods disclosed in U.S. Pat. Nos. 4,666,844; 5,350,688; 6,153,813; 6,333,449; 6,288,312; 6,365,807; 6,329,571, and the like. Other plant transformation methods are disclosed, for example, in U.S. Pat. Nos. 5,932,782; 6,153,811; 6,140,553; 5,969,213; 6,020,539, and the like. Any plant transformation method suitable for inserting a transgene into a particular plant may be used in accordance with the invention.
[0063] According to the present invention, the introduced polynucleotide may be maintained in the plant cell stably if it is incorporated into a non-chromosomal autonomous replicon or integrated into the plant chromosomes. Alternatively, the introduced polynucleotide may be present on an extra-chromosomal non-replicating vector and may be transiently expressed or transiently active.
[0064] Another aspect of the invention pertains to an isolated polypeptide having a sequence selected from the group consisting of the polypeptide sequences listed in Table 1. An "isolated" or "purified" polypeptide is free of some of the cellular material when produced by recombinant DNA techniques, or chemical precursors or other chemicals when chemically synthesized. The language "substantially free of cellular material" includes preparations of a polypeptide in which the polypeptide is separated from some of the cellular components of the cells in which it is naturally or recombinantly produced. In one embodiment, the language "substantially free of cellular material" includes preparations of a polypeptide of the invention having less than about 30% (by dry weight) of contaminating polypeptides, more preferably less than about 20% of contaminating polypeptides, still more preferably less than about 10% of contaminating polypeptides, and most preferably less than about 6% contaminating polypeptides.
[0065] The determination of activities and kinetic parameters of enzymes is well established in the art. Experiments to determine the activity of any given altered enzyme must be tailored to the specific activity of the wild-type enzyme, which is well within the ability of one skilled in the art. Overviews about enzymes in general, as well as specific details concerning structure, kinetics, principles, methods, applications and examples for the determination of many enzyme activities are abundant and well known to one skilled in the art.
[0066] The invention is also embodied in a method of producing a transgenic plant comprising at least one polynucleotide listed in Table 1, wherein expression of the polynucleotide in the plant results in the plant's increased growth and/or yield under normal or water-limited conditions and/or increased tolerance to an environmental stress as compared to a wild type variety of the plant comprising the steps of: (a) introducing into a plant cell an expression vector comprising at least one polynucleotide listed in Table 1, and (b) generating from the plant cell a transgenic plant that expresses the polynucleotide, wherein expression of the polynucleotide in the transgenic plant results in the plant's increased growth and/or yield under normal or water-limited conditions and/or increased tolerance to environmental stress as compared to a wild type variety of the plant. The plant cell may be, but is not limited to, a protoplast, gamete producing cell, and a cell that regenerates into a whole plant. As used herein, the term "transgenic" refers to any plant, plant cell, callus, plant tissue, or plant part, that contains at least one recombinant polynucleotide listed in Table 1. In many cases, the recombinant polynucleotide is stably integrated into a chromosome or stable extra-chromosomal element, so that it is passed on to successive generations.
[0067] The present invention also provides a method of increasing a plant's growth and/or yield under normal or water-limited conditions and/or increasing a plant's tolerance to an environmental stress comprising the steps of increasing the expression of at least one polynucleotide listed in Table 1 in the plant. Expression of a protein can be increased by any method known to those of skill in the art.
[0068] The effect of the genetic modification on plant growth and/or yield and/or stress tolerance can be assessed by growing the modified plant under normal and.or less than suitable conditions and then analyzing the growth characteristics and/or metabolism of the plant. Such analysis techniques are well known to one skilled in the art, and include dry weight, wet weight, polypeptide synthesis, carbohydrate synthesis, lipid synthesis, evapotranspiration rates, general plant and/or crop yield, flowering, reproduction, seed setting, root growth, respiration rates, photosynthesis rates, metabolite composition, etc., using methods known to those of skill in biotechnology.
[0069] The invention is further illustrated by the following examples, which are not to be construed in any way as imposing limitations upon the scope thereof.
EXAMPLE 1
Cloning of Full-Length cDNAs
[0070] The full-length DNA sequence of the Physcomitrella patens EST65 methionine sulfoxide reductase family protein (SEQ ID NO:43) was blasted against proprietary databases of canola, soybean, rice, maize, linseed, sunflower, barley, and wheat cDNAS at an e value of e-10 (Altschul et al., 1997, Nucleic Acids Res. 25: 3389-3402). All the contig hits were analyzed for the putative full length sequences, and the longest clones representing the putative full length contigs were fully sequenced. One homolog from canola, two homologs from rice, one homolog from soybean, and one homolog from wheat were identified. The degrees of amino acid identity and similarity of these sequences to the respective closest known public sequences are indicated in Tables 2 through 6 (Align 2.0).
TABLE-US-00002 TABLE 2 Comparison of BN51364980 (SEQ ID NO: 2) to known methionine sulfoxide reductases Public Database Sequence Accession # Species Identity (%) NP_564640 Arabidopsis thaliana 82.00% AAM65202 A. thaliana 66.70% BAD35399 O. sativa 56.10% NP_001057620 O. sativa 55.80% ZP_01592095 Geobacter lovleyi SZ 42.40%
TABLE-US-00003 TABLE 3 Comparison of OS34096188 (SEQ ID NO: 4) to known methionine sulfoxide reductases Public Database Sequence Accession # Species Identity (%) ABE84787 Medicago truncatula 72.60% NP_567639 A. thaliana 61.90% AAM62876 A. thaliana 61.00% NP_567271 A. thaliana 58.70% EAY98001 O. sativa 57.80%
TABLE-US-00004 TABLE 4 Comparison of OS32583643 (SEQ ID NO: 6) to known methionine sulfoxide reductases Public Database Sequence Accession # Species Identity (%) NP_001057620 O. sativa 99.10% BAD35399 O. sativa 74.80% NP_564640 A. thaliana 59.60% AAM65202 A. thaliana 53.70% YP_846684 Syntrophobacter 44.20% fumaroxidans MPOB
TABLE-US-00005 TABLE 5 Comparison of GM53626178 (SEQ ID NO: 8) to known methionine sulfoxide reductases Public Database Sequence Accession # Species Identity (%) ABE84787 M. truncatula 72.80% NP_567639 A. thaliana 60.90% AAM62876 A. thaliana 60.90% EAY98001 O. sativa 59.10% AAO72582 O. sativa 57.60%
TABLE-US-00006 TABLE 6 Comparison of TA56540264 (SEQ ID NO: 10) to known methionine sulfoxide reductases Public Database Sequence Accession # Species Identity (%) EAY98001 O. sativa 72.90% NP_001055501 O. sativa 69.50% ABE84787 M. truncatula 65.50% NP_567639 A. thaliana 60.60% AAM62876 A. thaliana 59.60%
[0071] The full-length DNA sequence of the P. patens EST12 homeodomain leucine zipper protein (SEQ ID NO:44) was blasted against proprietary databases of canola, soybean, rice, maize, linseed, sunflower, barley, and wheat cDNAS at an e value of e-10 (Altschul et al., 1997, Nucleic Acids Res. 25: 3389-3402). All the contig hits were analyzed for the putative full length sequences, and the longest clones representing the putative full length contigs were fully sequenced. One homolog from canola, one homolog from soybean, and one homolog from wheat were identified. The degrees of amino acid identity and similarity of these sequences to the respective closest known public sequence are indicated in Tables 7 through 9 (Align 2.0).
TABLE-US-00007 TABLE 7 Comparison of BN45206322 (SEQ ID NO: 12) to known homeodomain leucine zipper proteins Public Database Sequence Accession # Species Identity (%) AAR04932 B. napus 95.20% AAF73482 Brassica rapa 91.30% AAD41726 A. thaliana 81.20% NP_195716 A. thaliana 69.90% AAK96762 A. thaliana 69.60%
TABLE-US-00008 TABLE 8 Comparison of GM48923793 (SEQ ID NO: 14) to known homeodomain leucine zipper proteins Public Database Sequence Accession # Species Identity (%) AAX98G70 G. e max 56.00% AAK84886 Phaseolus vulgaris 51.60% CAA64417 Solanum lycopersicum 47.40% BAA05624 Daucus carota 46.70% AAF01765 G. max 44.00%
TABLE-US-00009 TABLE 9 Comparison of TA55969932 (SEQ ID NO: 16) to known homeodomain leucine zipper proteins Public Database Sequence Accession # Species Identity (%) NP_001048008 O. sativa 74.00% EAY87390 O. sativa 73.80% NP_001061807 O. sativa 49.90% AAD37698 O. saliva 49.60% AAS83417 O. sativa 46.30%
[0072] The full-length DNA sequence of the P. patens EST307 A20 and AN1 domain containing zinc finger protein (SEQ ID NO:45) was blasted against proprietary databases of canola, soybean, rice, maize, linseed, sunflower, barley, and wheat cDNAS at an e value of e-10 (Altschul et al., 1997, Nucleic Acids Res. 25: 3389-3402). All the contig hits were analyzed for the putative full length sequences, and the longest clones representing the putative full length contigs were fully sequenced. Two homologs from canola, one homolog from barley, two homologs from corn, two homologs from linseed, three homolog from soybean, three homologs from rice, and one homolog from wheat were identified. The degrees of amino acid identity and similarity of these sequences to the respective closest known public sequences are indicated in Tables 10 through 22 (Align 2.0).
TABLE-US-00010 TABLE 10 Comparison of BN47310186 (SEQ ID NO: 18) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) NP_564585 A. thaliana 88.70% AAN71995 A. thaliana 88.10% ABL67658 Citrus cv. Shiranuhi 59.40% AAQ84334 O. sativa 56.00% AAD38146 Prunus armeniaca 55.20%
TABLE-US-00011 TABLE 11 Comparison of BN51359456 (SEQ ID NO: 20) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) NP_190848 A. thaliana 71.60% AAK68811 A. thaliana 71.10% NP_565844 A. thaliana 66.00% ABE93196 M. truncatula 51.10% AAN71995 A. thaliana 47.10%
TABLE-US-00012 TABLE 12 Comparison of HV62552639 (SEQ ID NO: 22) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) NP_001055132 O. sativa 70.10% AAR96005 Musa acuminata 51.80% AAA33773 P. vulgaris 42.60% EAZ09556 O. sativa 40.80% EAZ45178 O. sativa 39.40%
TABLE-US-00013 TABLE 13 Comparison of ZM61995511 (SEQ ID NO: 24) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) AAQ84334 O. sativa 79.50% AAX14637 Z. mays 77.00% EAZ01657 O. sativa 71.70% ABL67658 Citrus cv. Shiranuhi 69.50% NP_001046186 O. sativa 65.90%
TABLE-US-00014 TABLE 14 Comparison of LU61567101 (SEQ ID NO: 26) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) CAE73100 Caenorhabditis 29.30% briggsae NP_190848 A. thaliana 29.20% XP_001357850 Drosophila 29.20% pseudoobscura EAY92150 O. sativa 29.10% ABL97956 B. rapa 28.90%
TABLE-US-00015 TABLE 15 Comparison of LU61893412 (SEQ ID NO: 28) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) ABL67658 Citrus cv. Shiranuhi 67.40% AAD38146 P. armeniaca 64.60% AAQ84334 O. sativa 61.70% ABN08135 M. truncatula 61.50% AAN71995 A. thaliana 61.00%
TABLE-US-00016 TABLE 16 Comparison of OS39781852 (SEQ ID NO: 30) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) EAZ45178 O. sativa 99.40% EAZ09556 O. sativa 99.40% NP_001063521 O. sativa 65.90% ABI23728 Chrysanthemum × 58.80% morifolium AAA33773 P. vulgaris 47.30%
TABLE-US-00017 TABLE 17 Comparison of OS34701560 (SEQ ID NO: 32) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) NP_565844 A. thaliana 58.80% ABE93196 M. truncatula 56.40% NP_190848 A. thaliana 55.60% AAK68811 A. thaliana 55.60% ABL67658 Citrus cv. Shiranuhi 48.30%
TABLE-US-00018 TABLE 18 Comparison of OS36821256 (SEQ ID NO: 34) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) EAZ45178 O. sativa 63.00% EAZ09556 O. sativa 63.00% ABI23728 Chrysanthemum × 51.10% morifolium AAA33773 P. vulgaris 43.40% AAX14637 Z. mays 43.30%
TABLE-US-00019 TABLE 19 Comparison of GM51659494 (SEQ ID NO: 36) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) AAA33773 P. vulgaris 58.00% EAZ09556 O. sativa 54.20% EAZ45178 O. sativa 54.20% NP_566429 A. thaliana 51.50% ABI23728 Chrysanthemum × 44.40% morifolium
TABLE-US-00020 TABLE 20 Comparison of GM49780101 (SEQ ID NO: 38) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) ABN08135 M. truncatula 76.70% ABL67658 Citrus cv. Shiranuhi 66.30% AAD38146 P. armeniaca 64.60% AAQ84334 O. sativa 64.40% AAX14637 Z. mays 61.90%
TABLE-US-00021 TABLE 21 Comparison of GM59637305 (SEQ ID NO: 40) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) AAD38146 P. armeniaca 69.30% ABL67658 Citrus cv. Shiranuhi 68.60% AAQ84334 O. sativa 65.90% AAX14637 Z. mays 64.00% NP_564585 A. thaliana 63.70%
TABLE-US-00022 TABLE 22 Comparison of TA55974113 (SEQ ID NO: 42) to known A20 and AN1 domain containing zinc finger proteins Public Database Sequence Accession # Species Identity (%) EAZ09556 O. sativa 73.20% EAZ45178 O. sativa 72.60% ABI23728 Chrysanthemum × 54.20% morifolium NP_001063521 O. sativa 49.00% AM33773 P. vulgaris 46.10%
EXAMPLE 2
Water Stress-Tolerant Arabidopsis Plants
[0073] The polynucleotides of Table 1 are ligated into a binary vector containing a selectable marker. The resulting recombinant vector contains the corresponding gene in the sense orientation under a constitutive promoter. The recombinant vectors are transformed into an Agrobacterium tumefaciens strain according to standard conditions. A. thaliana ecotype Col-0 or C24 are grown and transformed according to standard conditions. T1 and T2 plants are screened for resistance to the selection agent conferred by the selectable marker gene. T3 seeds are used in greenhouse or growth chamber experiments.
[0074] Approximately 3-5 days prior to planting, seeds are refrigerated for stratification. Seeds are then planted, fertilizer is applied and humidity is maintained using transparent domes. For the "biomass" assay, plants are grown in a greenhouse at 22° C. with photoperiod of 16 hours light/8 hours dark and watered twice a week. For the "cycling drought" assay, plants are grown in a growth chamber at 22° C. with 55% relative humidity with photoperiod was set at 16 h light/8 h dark and watered on days 0, 18, and 25 after sowing.
[0075] At 19 and 22 days, plant area, leaf area, biomass, color distribution, color intensity, and growth rate for each plant are measured using using a commercially available imaging system. Biomass is calculated as the total plant leaf area at the last measuring time point. Growth rate is calculated as the plant leaf area at the last measuring time point minus the plant leaf area at the first measuring time point divided by the plant leaf area at the first measuring time point. Health Index is calculated as the dark green leaf area divided by the total plant leaf area. Table 23 presents the biomass, growth rate, and health index for independent transformation events (lines) of transgenic plants overexpressing polynucleotides represented as SEQ ID NOs:5, 7,and 13. The percent change of a line compared to pooled wild-type controls was calculated, and the significant (p) value was calculated using a t-test. An event was called positive if percent change was greater than 0 and p<11, not significant (NS) if p>0.1 regardless of percent change, and negative if percent change was less than 0 and p<0.1.
TABLE-US-00023 TABLE 23 Number of events called in each Result result category per assay Gene ID catagory Biomass Growth Rate Health Index OS32583643 Positive 1 2 2 SEQ ID NO: 5 NS 8 5 6 negative 1 3 2 GM53626178 Positive 6 2 1 SEQ ID NO: 7 NS 3 6 7 negative 1 2 2 GM48923793 Positive 3 1 4 SEQ ID NO: 13 NS 2 2 4 negative 5 7 2
EXAMPLE 3
Nitrogen Stress Tolerant Arabidopsis Plants
[0076] The polynucleotides of Table 1 are ligated into a binary vector containing a selectable marker. The resulting recombinant vector contains the corresponding gene in the sense orientation under a constitutive promoter. The recombinant vectors are transformed into an A. tumefaciens strain according to standard conditions. A. thaliana ecotype Col-0 or C24 are grown and transformed according to standard conditions. T1 and T2 plants are screened for resistance to the selection agent conferred by the selectable marker gene.
[0077] Plants are grown in flats using a substrate that contains no organic components. Each fiat is wet with water before seedlings resistant to the selection agent are transplanted onto substrate. Plants are grown in a growth chamber set to 22° C. with a 55% relative humidity with photoperiod set at 16 h light/8 h dark. A controlled low or high nitrogen nutrient solution is added to waterings on Days 12, 15, 22 and 29. Watering without nutrient solution occurs on Days 18, 25, and 32. Images of all plants in a tray are taken on days 26, 30, and 33 using a commercially available imaging system. At each imaging time point, biomass and plant phenotypes for each plant are measured including plant area, leaf area, biomass, color distribution, color intensity, and growth rate.
EXAMPLE 4
Stress-Tolerant Rapeseed/Canola Plants
[0078] Canola cotyledonary petioles of 4 day-old young seedlings are used as explants for tissue culture and transformed according to EP1566443. The commercial cultivar Westar (Agriculture Canada) is the standard variety used for transformation, but other varieties can be used. A. tumefaciens GV3101:pMP90RK containing a binary vector is used for canola transformation. The standard binary vector used for transformation is pSUN (WO02/00900), but many different binary vector systems have been described for plant transformation (e.g. An, G. in Agrobacterium Protocols, Methods in Molecular Biology vol 44, pp 47-62, Gartland K M A and M R Davey eds. Humana Press, Totowa, N.J.). A plant gene expression cassette comprising a selection marker gene and a plant promoter regulating the transcription of the cDNA encoding the polynucleotide is employed. Various selection marker genes can be used including the mutated acetohydroxy acid synthase (AHAS) gene disclosed in U.S. Pat. Nos. 5,767,366 and 6,225,105. A suitable promoter is used to regulate the trait gene to provide constitutive, developmental, tissue or environmental regulation of gene transcription.
[0079] Canola seeds are surface-sterilized in 70% ethanol for 2 min, incubated for 15 min in 55° C. warm tap water and then in 1.5% sodium hypochlorite for 10 minutes, followed by three rinses with sterilized distilled water. Seeds are then placed on MS medium without hormones, containing Gamborg B5 vitamins, 3% sucrose, and 0.8% Oxoidagar. Seeds are germinated at 24° C. for 4 days in low light (<50 μMol/m2s, 16 hours light). The cotyledon petiole explants with the cotyledon attached are excised from the in vitro seedlings, and inoculated with Agrobacterium by dipping the cut end of the petiole explant into the bacterial suspension. The explants are then cultured for 3 days on MS medium including vitamins containing 3.75 mg/l BAP, 3% sucrose, 0.5 g/l MES, pH 5.2, 0.5 mg/l GA3, 0.8% Oxoidagar at 24° C., 16 hours of light. After three days of co-cultivation with Agrobacterium, the petiole explants are transferred to regeneration medium containing 3.75 mg/l BAP, 0.5 mg/l GA3, 0.5 g/l MES, pH 5.2, 300 mg/l timentin and selection agent until shoot regeneration. As soon as explants start to develop shoots, they are transferred to shoot elongation medium (A6, containing full strength MS medium including vitamins, 2% sucrose, 0.5% Oxoidagar, 100 mg/l myo-inositol, 40 mg/l adenine sulfate, 0.5 g/l MES, pH 5.8, 0.0025 mg/l BAP, 0.1 mg/l IBA, 300 mg/l timentin and selection agent).
[0080] Samples from both in vitro and greenhouse material of the primary transgenic plants (T0) are analyzed by qPCR using TaqMan probes to confirm the presence of T-DNA and to determine the number of T-DNA integrations.
[0081] Seed is produced from the primary transgenic plants by self-pollination. The second-generation plants are grown in greenhouse conditions and self-pollinated. The plants are analyzed by qPCR using TaqMan probes to confirm the presence of T-DNA and to determine the number of T-DNA integrations. Homozygous transgenic, heterozygous transgenic and azygoes (null transgenic) plants are compared for their stress tolerance, for example, in assays similar to those described in Examples 2 and 3, and for yield, both in the greenhouse and in field studies.
EXAMPLE 5
Screening for Stress-Tolerant Rice Plants
[0082] Transgenic rice plants comprising a polynucleotide of Table 1 are generated using known methods. Approximately 15 to 20 independent transformants (T0) are generated. The primary transformants are transferred from tissue culture chambers to a greenhouse for growing and harvest of T1 seeds. Five events of the T1 progeny segregated 3:1 for presence/absence of the transgene are retained. For each of these events, 10 T1 seedlings containing the transgene (hetero- and homozygotes), and 10 T1 seedlings lacking the transgene (nullizygotes) are selected by visual marker screening. The selected T1 plants are transferred to a greenhouse. Each plant receives a unique barcode label to link unambiguously the phenotyping data to the corresponding plant. The selected T1 plants are grown on soil in 10 cm diameter pots under the following environmental settings: photoperiod=11.5 h, daylight intensity=30,000 lux or more, daytime temperature=28° C. or higher, night time temperature=22° C., relative humidity=60-70%. Transgenic plants and the corresponding nullizygotes are grown side-by-side at random positions. From the stage of sowing until the stage of maturity, the plants are passed several times through a digital imaging cabinet. At each time point digital, images (2048×1536 pixels, 16 million colours) of each plant are taken from at least 6 different angles.
[0083] The data obtained in the first experiment with T1 plants are confirmed in a second experiment with T2 plants. Lines that have the correct expression pattern are selected for further analysis. Seed batches from the positive plants (both hetero- and homozygotes) in T1 are screened by monitoring marker expression. For each chosen event, the heterozygote seed batches are then retained for T2 evaluation. Within each seed batch, an equal number of positive and negative plants are grown in the greenhouse for evaluation.
[0084] Transgenic plants are screened for their improved growth and/or yield and/or stress tolerance, for example, using assays similar to those described in Examples 2 and 3, and for yield, both in the greenhouse and in field studies.
EXAMPLE 6
Stress-Tolerant Soybean Plants
[0085] The polynucleotides of Table 1 are transformed into soybean using the methods described in commonly owned copending international application number WO 2005/121345, the contents of which are incorporated herein by reference.
[0086] The transgenic plants generated are then screened for their improved growth under water-limited conditions and/or drought, salt, and/or cold tolerance, for example, using assays similar to those described in Examples 2 and 3, and for yield, both in the greenhouse and in field studies.
EXAMPLE 7
Stress-Tolerant Wheat Plants
[0087] The polynucleotides of Table 1 are transformed into wheat using the method described by Ishida et al., 1996, Nature Biotech. 14745-50. Immature embryos are co-cultivated with Agrobacterium tumefaciens that carry "super binary" vectors, and transgenic plants are recovered through organogenesis. This procedure provides a transformation efficiency between 2.5% and 20%. The transgenic plants are then screened for their improved growth and/or yield under water-limited conditions and/or stress tolerance, for example, in assays similar to those described in Examples 2 and 3, and for yield, both in the greenhouse and in field studies.
EXAMPLE 8
Stress-Tolerant Corn Plants
[0088] The polynucleotides of Table 1 are transformed into immature embryos of corn using Agrobacterium. After imbibition, embryos are transferred to medium without selection agent, Seven to ten days later, embryos are transferred to medium containing selection agent and grown for 4 weeks (two 2-week transfers) to obtain transformed callus cells. Plant regeneration is initiated by transferring resistant calli to medium supplemented with selection agent and grown under light at 25-27° C. for two to three weeks. Regenerated shoots are then transferred to rooting box with medium containing selection agent. Plantiets with roots are transferred to potting mixture in small pots in the greenhouse and after acclimatization are then transplanted to larger pots and maintained in greenhouse till maturity.
[0089] Each of these plants is uniquely labeled, sampled and analyzed for transgene copy number. Transgene positive and negative plants are marked and paired with similar sizes for transplanting together to large pots. This provides a uniform and competitive environment for the transgene positive and negative plants. The large pots are watered to a certain percent-age of the field water capacity of the soil depending on the severity of water-stress desired. The soil water level is maintained by watering every other day. Plant growth and physiology traits such as height, stem diameter, leaf rolling, plant wilting, leaf extension rate, leaf water status, chlorophyll content and photosynthesis rate are measured during the growth period. After a period of growth, the above ground portion of the plants is harvested, and the fresh weight and dry weight of each plant are taken. A comparison of the drought tolerance phenotype between the transgene positive and negative plants is then made.
[0090] The pots are covered with caps that permit the seedlings to grow through but minimize water loss. Each pot is weighed periodically and water added to maintain the initial water content. At the end of the experiment, the fresh and dry weight of each plant is measured, the water consumed by each plant is calculated and WUE of each plant is computed. Plant growth and physiology traits such as WUE, height, stem diameter, leaf rolling, plant wilting, leaf extension rate, leaf water status, chlorophyll content and photosynthesis rate are measured during the experiment. A comparison of WUE phenotype between the transgene positive and negative plants is then made.
[0091] These pots are kept in an area in the greenhouse that has uniform environmental conditions, and cultivated optimally. Each of these plants is uniquely labeled, sampled and analyzed for transgene copy number. The plants are allowed to grow under theses conditions until they reach a predefined growth stage. Water is then withheld. Plant growth and physiology traits such as height, stem diameter, leaf rolling, plant wilting, leaf extension rate, leaf water status, chlorophyll content and photosynthesis rate are measured as stress intensity increases. A comparison of the dessication tolerance phenotype between transgene positive and negative plants is then made.
[0092] Segregating transgenic corn seeds for a transformation event are planted in small pots for testing in a cycling drought assay. These pots are kept in an area in the greenhouse that has uniform environmental conditions, and cultivated optimally. Each of these plants is uniquely labeled, sampled and analyzed for transgene copy number. The plants are allowed to grow under theses conditions until they reach a predefined growth stage. Plants are then repeatedly watered to saturation at a fixed interval of time. This water/drought cycle is repeated for the duration of the experiment. Plant growth and physiology traits such as height, stem diameter, leaf rolling, leaf extension rate, leaf water status, chlorophyll content and photosynthesis rate are measured during the growth period. At the end of the experiment, the plants are harvested for above-ground fresh and dry weight. A comparison of the cycling drought tolerance phenotype between transgene positive and negative plants is then made.
[0093] In order to test segregating transgenic corn for drought tolerance under rain-free conditions, managed-drought stress at a single location or multiple locations is used. Crop water availability is controlled by drip tape or overhead irrigation at a location which has less than 10 cm rainfall and minimum temperatures greater than 5° C. expected during an average 5 month season, or a location with expected in-season precipitation intercepted by an automated "rainout shelter" which retracts to provide open field conditions when not required. Standard agronomic practices in the area are followed for soil preparation, planting, fertilization and pest control. Each plot is sown with seed segregating for the presence of a single transgenic insertion event. A Taqman transgene copy number assay is used on leaf samples to differentiate the transgenics from null-segregant control plants. Plants that have been genotyped in this manner are also scored for a range of phenotypes related to drought-tolerance, growth and yield. These phenotypes include plant height, grain weight per plant, grain number per plant, ear number per plant, above ground dry-weight, leaf conductance to water vapor, leaf CO2 uptake, leaf chlorophyll content, photosynthesis-related chlorophyll fluorescence parameters, water use efficiency, leaf water potential, leaf relative water content, stem sap flow rate, stem hydraulic conductivity, leaf temperature, leaf reflectance, leaf light absorptance, leaf area, days to flowering, anthesis-silking interval, duration of grain fill, osmotic potential, osmotic adjustment, root size, leaf extension rate, leaf angle, leaf rolling and survival. All measurements are made with commercially available instrumentation for field physiology, using the standard protocols provided by the manufacturers. Individual plants are used as the replicate unit per event.
[0094] In order to test non-segregating transgenic corn for drought tolerance under rainfree conditions, managed-drought stress at a single location or multiple locations is used. Crop water availability is controlled by drip tape or overhead irrigation at a location which has less than 10 cm rainfall and minimum temperatures greater than 5° C. expected during an average 5 month season, or a location with expected in-season precipitation intercepted by an automated "rain-out shelter" which retracts to provide open field conditions when not required. Standard agronomic practices in the area are followed for soil preparation, planting, fertilization and pest control. Trial layout is designed to pair a plot containing a non-segregating transgenic event with an adjacent plot of null-segregant controls. A null segregant is progeny (or lines derived from the progeny) of a transgenic plant that does not contain the transgene due to Mendelian segregation. Additional replicated paired plots for a particular event are distributed around the trial. A range of phenotypes related to drought-tolerance, growth and yield are scored in the paired plots and estimated at the plot level. When the measurement technique could only be applied to individual plants, these are selected at random each time from within the plot. These phenotypes include plant height, grain weight per plant, grain number per plant, ear number per plant, above ground dry-weight, leaf conductance to water vapor, leaf CO2 uptake, leaf chlorophyll content, photosynthesis-related chlorophyll fluorescence parameters, water use efficiency, leaf water potential, leaf relative water content, stem sap flow rate, stem hydraulic conductivity, leaf temperature, leaf reflectance, leaf light absorptance, leaf area, days to flowering, anthesis-silking interval, duration of grain fill, osmotic potential, osmotic adjustment, root size, leaf extension rate, leaf angle, leaf rolling and survival. All measurements are made with commercially available instrumentation for field physiology, using the standard protocols provided by the manufacturers. Individual plots are used as the replicate unit per event.
[0095] To perform multi-location testing of transgenic corn for drought tolerance and yield, five to twenty locations encompassing major corn growing regions are selected. These are widely distributed to provide a range of expected crop water availabilities based on average temperature, humidity, precipitation and soil type. Crop water availability is not modified beyond standard agronomic practices. Trial layout is designed to pair a plot containing a non-segregating transgenic event with an adjacent plot of null-segregant controls. A range of phenotypes related to drought-tolerance, growth and yield are scored in the paired plots and estimated at the plot level. When the measurement technique could only be applied to individual plants, these are selected at random each time from within the plot. These phenotypes included plant height, grain weight per plant, grain number per plant, ear number per plant, above ground dry-weight, leaf conductance to water vapor, leaf CO2 uptake, leaf chlorophyll content, photosynthesis-related chlorophyll fluorescence parameters, water use efficiency, leaf water potential, leaf relative water content, stem sap flow rate, stem hydraulic conductivity, leaf temperature, leaf reflectance, leaf light absorptance, leaf area, days to flowering, anthesis-silking interval, duration of grain fill, osmotic potential, osmotic adjustment, root size, leaf extension rate, leaf angle, leaf rolling and survival. All measurements are made with commercially available instrumentation for field physiology, using the standard protocols provided by the manufacturers. Individual plots are used as the replicate unit per event.
TABLE-US-00024 APPENDIX cDNA sequence of BN51364980 for canola (SEQ ID NO: 1): atggcttcttctagttgtttcaccattcagtcacgtttcgtctcagcgagaacaaagctcgattcaatctccaa- accgagtctctccggattc gcttgtcgttctcttacaaaacccagaaacttgaatctctctgttcttcttcggtgttccatgggttcctttaa- ctcttctcagaaatcagacaac gtccaagaagctgcaaagagtgactttgcttcaataagtgaaggtgagtggaagaaacggctaacaccagaaca- gtattacatcac cagacagaagggaacagagagagctttcactggtgagtattggaatacaaagaccccaggagtatacaaatgta- tctgttgcgaca cgccactgtttgactcatcaacaaagcttgatagtggaaccgggtggccatcgtattaccaacctattggaaac- aatgtgaagtcaaa gctggacctctctatcatcttcatgcctagacaagaagttatctgtgctgtttgtaacgcccatcttggtcatg- tcttcgatgacggtccacg accaaccggaaaacgatattgcctcaacagtgctgctctgaaacttgagtcattggag agaacaagagaatga The BN51364980 cDNA is translated into the following amino acid sequence (SEQ ID NO: 2): massscftiqsrfvsartkldsiskpslsgfacrsltkprnlnlsvllrcsmgsfnssqksdnvqeaaksdfas- isegewkkrltpeqyyitr qkgteraftgeywntktpgvykciccdtplfdsstkldsgtgwpsyyqpignnvkskldlsiifmprqevicav- cnahlghvfddgprptg kryclnsaalkleslertre cDNA sequence of OS34096188 from rice (SEQ ID NO: 3): atgggcttcaatattctgagaaccacttccatctccactcctatctcttcctccaaatccaaacccattttctc- aactcttcttcgttcttctccttc caccattttccccccaaagtccgttactcccaccactcttttcgtttctgccacccccttcttcactctccatc- ccaagcttggttttcgtggtgg gattgtggccatggccgcacctggctctctccgcaaatccgaggaagagtggcgcgcaattctctcccctgaac- agtttcggatcctca ggcaaaagggcaccgagttccctggaacaggagagtatgacaagttctatgaagagggagtttacaactgtgct- ggttgtgggactc cactctacaggtccataacaaaattcaattctggttgtggctggccagccttctatgaggggattcccggagcc- ataaatcgcaatccg gatcctgatgggatgaggacagaaataacgtgtgctgcttgtgggggacatctaggtcacgtctttaaaggaga- aggatttccaacac ccactaacgaacgccattgtgtcaatagcatttcgctgaaatttgcgccagccaattcttattcttaa The OS34096188 cDNA is translated into the following amino acid sequence (SEQ ID NO: 4): mgfnilrttsistpissskskpifstllrsspstifppksvtpttlfvsatpfftlhpklgfrggivamaapgs- lrkseeewrailspeqfrilrqkg tefpgtgeydkfyeegvyncagcgtplyrsitkfnsgcgwpafyegipgainrnpdpdgmrteitcaacgghlg- hvfkgegfptptnerhc vnsislkfapansys cDNA sequence of OS32583643 from rice (SEQ ID NO: 5): atggccatgcggcaatacgcggctgctaccgctgcctcctccagtttcagagcacgtccacgggcgcgcccctc- ctgcctcccagcc gccgccctgcccttggcgccttgctgtggtgtggcgtggagccgtgctagctacaggcgagcctccgttcgtgc- catgggtgccgcttc atcgtcttcgtcgtcgtcgtcgtcgtctccgtcgccgcagggtcaagcccaagcccaagcccaaggtaaaccga- actacagtacatct ctgactgatgaggagtggaggaagcgcctgacaaaagatcagtattacattactcggcagaagggcacagaaag- agcatttactg gggaatactggaacaccaaaaccccgggcatctaccattgtgtctgctgtgacacccctctttttgagtcatcg- accaaatttgatagtg gtactgggtggccgtcatattatcaacccattggagataatgtaaagtgcaagcttgatatgtccatcatattc- atgcctcggactgaggt gctgtgtgctgtctgtgacgctcatctggggcacgtgtttgatgatgggccacgaccaacagggaaaagatact- gtatcaatagcgcat ctctcaagctgaagaagacccagtag The OS32583643 cDNA is translated into the following amino acid sequence (SEQ ID NO: 6): mamrqyaaataasssfrarprarpsclpaaalplapccgvawsrasyrrasvramgaasssssssssspspqgq- aqaqaqgkp nystsltdeewrkrltkdqyyitrqkgteraftgeywntktpglyhcvccdtplfesstkfdsgtgwpsyyqpl- gdnvkckldmslifmprt evlcavcdahlghvfddgprptgkrycinsaslklkktq cDNA sequence of GM53626178 from soybean (SEQ ID NO: 7): atgggattgagtattctgagaagcacttccatttccactcctatctcttcctccaaatccaaacccattttctc- aactcttgttcgttcatctttcg cctccatttcccccacaaagtgtgttactcccaccactcttttcgtttctgccacccccttcttcaccgcctca- cccaagcgcggctttcgtgg tgggattgtggccatggccgccgctggctcgctccgcaaatcagaggaagagtggcgcgcagttctctcccctg- aacagtttcgtattct caggcaaaagggcaccgagttccctggaacaggagagtatgacaagttctttgatgagggagtttacaactgtg- ctggttgtgggac acctctctacaggtccttaacaaaattcaattctggttgtggctggccagccttctatgaggggattcctggag- ccataaatcgcaatccg gaccctgatgggatgaggacagaaataacgtgtgctgcttgtgggggacatctaggtcacgtctttaaaggaga- aggatttccaacgc ccactaacgaacgccattgtgtcaatagcatttcactgaaatttgcgccagccaattcttaa The GM53626178 cDNA is translated into the following amino acid sequence (SEQ ID NO: 8): mglsilrstststpissskskpifstlvrssfasisptkcvtpttlfvsatpfftaspkrgfrggivamaaags- lrkseeewravlspeqfrilrqk gtefpgtgeydkffdegvyncagcgtplyrsltkfnsgcgwpafyegipgalnrnpdpdgmrteitcaacgghl- ghvlkgegfptptne rhcvnsislkfapans cDNA sequence of TA56540264 from wheat (SEQ ID NO: 9): atggcgtcgccccacgcccacccggccacgcggcccctctcatcgctcccgtccctcctcctcgcccgctcctc- ctccgccgccaccg ccgccgcgtcgtccgcccgccccgcctccctctccctctcgtgctcgcggtcgcgggcgcgggcctactgccca- gccggacgacggt tgccgggcgccgtggtggctatgtcgtcggcggcgcccacgccggggcccgtgcagaagtcggaggaggagtgg- gaggccgtcc tcacgccggagcagttccgcatcctccgccgcaagggcaccgagtatcctggaacaggtgaatatgacaagttc- ttcagtgagggta tttacggatgtgctggctgtggaacccccttgtacaaatcatctacgaagttcaactcagggtgtggttggcca- gcattctatgaaggattt cctggagccataaaacggacggcggatcctgatgggaggcgaattgagatcacatgtgctgcttgtgaaggaca- tctggggcatgtg ttcaaaggggaggggttcaacacgccgactgatgagcgacactgcgtcaacagtatctcactcaagttcgttcc- ggcctctgaagag gctagttga The TA 56540264 cDNA is translated into the following amino acid sequence (SEQ ID NO: 10) masphahpatrplsslpslllarsssaataaassarpaslslscsrsraraycpagrrlpgavvamssaaptpg- pvqkseeeweavlt peqfrilrrkgteypgtgeydkffsegiygcagcgtplyksstkfnsgcgwpafyegfpgaikrtadpdgrrie- itcaaceghlghvfkge gfntptderhcvnsislkfvpaseeas cDNA sequence of BN45206322 from canola (SEQ ID NO: 11) atgatgaagagattaagcagttcagattcagtgggtggtctcatctctttatgtcccactacttccacagatca- gccgaatccaagaaga tgcgggagagaatttcagtcgatgctcgaaggttacgaggaggaagaagaagaagccataaccgaggaaagagg- acaaaccg gtttagccgagaagaagagacggttaaacattaaccaagttaaagccttggagaaaaatttcgagttagagaac- aagcttgagcctg agaggaaagtgaagttagctcaagaacttggtctccaacctcgtcaagtagctgtttggtttcagaaccgccgt- gcgcggtggaagac aaaacagcttgagaaagattacggtgttctcaaaacgcaatacgattctctccgccataactttgattccctcc- gccgtgaaaatgaatc tcttcttcaagagatcggtaaactaaaagctaagcttaacggagaagaagaaggagatgatgttgatgaagaag- agaacaacttgg cgacgatggagagtgatgtttccgtcaaggaagaagaagtttcgttgccggagcagatcacagagccgccgtct- tctcctccgcagct tctagagcattccgacagtttcaattaccggagtttcaccgacctccgcgaccttcttccgttaaaggccgcgg- cttcctccgtcgccgcc gctggatcgtcggacagtagcgattcgagcgccgtgttgaacgaggaaagtagctctaacgttacggcggctcc- ggcgacggttccc ggcggcagtttcttgcagtttgtgaaaatggagcagacggaggatcacgacgactttctgagtggagaagaagc- gtgcgggtttttctc cgatgaacagccaccgtctctgcactggtattccaccgttgatcagtggaactga The BN45206322 cDNA is translated into the following amino acid sequence (SEQ ID NO: 12): mmkrlsssdsvgglislcpttstdqpnprrcgrefqsmlegyeeeeeeaiteergqtglaekkrrininqvkal- eknfelenkleperkv klaqelglqprqvavwfqnrrarwktkqlekdygvlktqydslrhnfdslrrenesllqeigklkaklngeeeg- ddvdeeennlatmesd vsvkeeevslpeqiteppssppqllehsdsfnyrsftdlrdllplkaaassvaaagssdssdssavlneesssn- vtaapatvpggsflqf vkmeqtedhddflsgeeacgffsdeqppslhwystvdqwn cDNA sequence of GM48923792 from soybean (SEQ ID NO: 13) atggcgggtagtggaagtgccttttccaacatcactagctttcttcgcacccaacaaccctcttctcaacctct- cgattcttctctcttcctctc tgcaccttcctctgctcctttcctcggttcgagatccatgatgagttttgatggagaaggagggaaggggtgta- acggctccttcttccgcg cgtttgacatggacgacaatggggatgagtgcatggacgagtactttcatcaacccgagaagaagcgacgtctc- tctgcgagccag gttcagtttctagagaagagcttcgaggaggagaacaagcttgaacccgagagaaagaccaaactagccaaaga- ccttggtttgca gccacggcaagttgctatttggttccagaaccgtagagctcggtggaagaacaaacagctggagaaggattacg- agactctgcatg caagttttgagaagtctcaagtccaactatgactgtcttctcaaggagaaagacaagttaaaagctgaggtggc- gagcctcactgagaa ggtgcttgcaagagggaaacaagaggggcacatgaagcaggctgaaagtgaaagtgaagaaacaaaaggattat- tgcatttgca ggaacaggaaccaccccagaggcttttactgcaatcagtttcggagggagaaggatccaaagtctcttctgtcg- ttgggggttgtaaa caggaagatatcagttcagcaaggagtgacattttggattcagatagtccacattacaccgatggagttcactc- tgcgctgctagagca tggtgattcttcttatgtgtttgagcctgatcaatcagatatgtcacaagatgaagaagataacctcagcaaga- gtctctacccttcgtacc tctttcccaaacttgaagaagatgtggattactccgacccacctgaaagttcttgtaattttggatttcctgag- gaagatcatgtcctttgga cctgggcttactactaa The GM48923793 cDNA is translated into the following amino acid sequence (SEQ ID NO: 14): magsgsafsnitsflrtqqpssqpldsslflsapssapflgsrsmmsfdgeggkgcngsffrafdmddngdecm- deyfhqpekkrrl sasqvqfleksfeeenkleperktklakdlglqprqvaiwfqnrrarwknkqlekdyetlhasfeslksnydcl- lkekdklkaevasltek vlargkqeghmkqaeseseetkgllhlqeqeppqrlllqsvsegegskvssvvggckqedissarsdlldsdsp- hytdgvhsalleh gdssyvfepdqsdmsqdeednlskslypsylfpkleedvdysdppesscnfgfpeedhvlwtwayy cDNA sequence of TA55969932 from wheat (SEQ ID NO: 15): atggagcccggccggctcatcttcaacacgtcgggctccggcaacggacagatgctcttcatggactgcggcgc- gggcggcatcgc cggcgcggccggcatgttccatcgaggggtgagaccggtcctcggcggcatggaagaagggcgcggcgtgaagc- ggcccttcttc acctcgccggatgacatgctggaggaggagtactacgacgagcagctcccggagaagaagcggcgcctcacgcc- ggagcaggt ccacctgctggagaggagcttcgaggaggagaacaagctggagccggagaggaagacggagctggcccgcaagc- tcgggctg cagccacggcaggtggccgtctggttccagaaccgccgcgcccggtggaagacaaagacgctggagcgcgactt- cgaccgcctc aaggcgtccttcgacgccctccgcgccgaccacgacgcgctcctccaggacaaccaccggctccggtcacaggt- ggtaacgttga ccgagaagatgcaagataaggaggcgccggaaggcagcttcggtgcagccgccgacgcctcggagccggagcag- gcggcgg cggaggcgaaggcttccttggccgacgccgaggagcaggccgcggcagcggaggcgttcgaggtggtgcagcag- cagctgcac gtgaaggacgaggagaggctgagcccggggagcggcgggagcgcggtgctggacgcgagggacgcgctgctcgg- gagcgga tgcggcctcgccggcgtggtggacagcagcgtggactcgtactgcttcccggggggcgccggcggcgacgagta- ccacgagtgcg tggtgggccccgtggcgggcggcatccagtcggaggaggacgacggcgcgggcagcgacgagggctgcagctac- taccccgac gacgccgccgtcttcttcgccgccgcgcaagggcacggccaccatcgcacggacgacgacgatcagcaggacga- cggccagat cagctactggatgtggaactag The TA55969932 cDNA is translated into the following amino acid sequence (SEQ ID NO: 16): mepgrlifntsgsgngqmlfmdcgaggiagaagmfhrgvrpvlggmeegrgvkrpfftspddmleeeyydeqlp- ekkrrltpeqvh llersfeeenkleperktelarklglqprqvavwfqnrrarwktktlerdfdrlkasfdalradhdallqdnhr- lrsqvvtltekmqdkeape gsfgaaadasepeqaaaeakasladaeeqaaaaeafevvqqqlhvkdeerlspgsggsavldardallgsgcgl- agvvdssvds ycfpggaggdeyhecvvgpvaggiqseeddgagsdegcsyypddaavffaaaqghghhrtddddqqddgqisyw- mwn cDNA sequence of BN47310186 from canola (SEQ ID NO: 17): atggaccacgacaaaacaggatgccaaagcccacctgaaggtcccaagctatgcatcaacaactgcggtttctt- cggaagcgctgc cacaatgaacatgtgttccaagtgtcacaaggctatcctgtttcaacaggaacagggggctaggtttgcatctg- cagtgtctggtggtac atcatcatccagcaacatcttaaaggaaacctttgctgctaccgcgctggttgatgctgaaaccaaatccgttg- agccggtggctgtctc tgtacagccatcttctgtccaagttgccgcagaggtagtagctccagaagccgctgcagcaaaactaaaggaag- gaccaagccgat gtgctacttgcaataaacgggttggtctgactggattcaaatgtcgctgtggtgacctcttctgcgggacgcac- cgttatgcagacataca caactgctccttcaattaccatgccgctgcgcaagaagctatagctaaagcaaacccggttgtgaaggcagaga- agcttgacaaaat ctga The BN47310186 cDNA is translated into the following amino acid sequence (SEQ ID NO: 18): mdhdktgcqsppegpklcinncgffgsaatmnmcskchkailfqqeqgarfasavsggtssssnilketfaata- lvdaetksvepva vsvqpssvqvaaevvapeaaaaklkegpsrcatcnkrvgltgfkcrcgdlfcgthryadihncsfnyhaaaqea-
lakanpvvkaekl dki cDNA sequence of BN51359456 from canola (SEQ ID NO: 19): atggcggaagagcatcgatgccagacgccggaaggccaccgtctctgtgctaacaactgcggcttcctcggcag- ctccgccaccat gaatctatgctccaactgctacggcgatctctgccttaagcaacagcaagcttccatgaaatccaccgtcgaat- cctctctctccgccgt atctcctccgtcgtcagagatcggctctatgcaatccaccgttgaatcctctctctccgacgtatctcctccat- caccggagaccatttcca tctcctctccaatgatccagcctctcgttcgaaacccatcagctgaattggaggtaacggcgacgaagacggtg- actccgccgccgg agcagcagcagaaacggccgaatcggtgcacgacgtgtaggaaacgggtcgggttgaccgggttcaagtgccgg- tgcgggacg actttttgcggggctcacaggtacccggaggtccatggatgcaccttcgatttcaaatcggccggtcgcgaaga- gatcgccaaggcg aacccactcgtcaaagcggcgaagcttcagaagatttga The BN51359456 cDNA is translated into the following amino acid sequence (SEQ ID NO: 20): maeehrcqtpeghrlcanncgflgssatmnlcsncygdlclkqqqasmkstvesslsavsppsseigsmqstve- sslsdvsppsp etisisspmiqplvrnpsaelevtatktvtpppeqqqkrpnrcttcrkrvgltgfkcrcgttfcgahrypevhg- ctfdfksagreelakanpl vkaaklqki cDNA sequence of HV62552639 from barley (SEQ ID NO: 21): atggcccaggagagttgtgatctcaacaaggacgaggccgagatcctgaagccatcctcctccacacaccttcg- cctccttcgccagcc acaccaccaccaccaaccgctcaaataccagaaccacaacctccacactcaccaccacaaccaccggcagctca- attcttgtcca ggccctgcgaggttgttcccatagagacttccaaaaagaggaaacatgctgatgcggtgtcaatggccattgtg- gttgagccattgtcg tctgtgctgttcgttaaccgttgcaacgtgtgccgcaagagagttggtttgaccgggttccgttgccggtgtga- gaagctcttttgtccgcgc caccggcattcagaaagccacgactgctcatttgattataaaactgtgggtcgggaggagattgcccgggcaaa- ccctctgatcagg gctgccaagatcattaggatatga The HV 62552639 cDNA is translated into the following amino acid sequence (SEQ ID NO: 22): maqescdlnkdeaeilkpssstpsppspatpppptaqipepqpphsppqppaaqflsrpcevvpletskkrkha- davsmalvvep lssvlfvnrcnvcrkrvgltgfrcrceklfcprhrhseshdcsfdyktvgreeiaranpliraakiiri cDNA sequence of ZM61995511 from corn (SEQ ID NO: 23): atggaacacaaggaggcgggctgccagcagccggagggcccaatcctatgcatcaataactgcggcttcttcgg- cagtgctgcga cgatgaacatgtgctccaagtgccacaaggagatgataacgaagcaggagcaggcccagctggctgcctccccc- atcgatagcat tgtcaatggcggtgacggcgggaaaggacctgtaattgctgcatctgtaaatgtggcagttcctcaagttgagc- agaagactattgttgt gcagcccatgcttgtagctgaaaccagcgaggctgctgctgtaatccccaaggccaaggaaggcccagaccggt- gcgcggcctgc aggaagcgtgttgggctgacgggatttagctgccgatgcgggaacatgtactgttcggtgcaccgctactccga- caaacatgactgtc agttcgactatcggactgcagcaagggacgcgattgccaaggccaatcctgtggtgagggcggagaagctcgac- aagatctga The ZM61995511 cDNA is translated into the following amino acid sequence (SEQ ID NO 24): mehkeagcqqpegpilcinncgffgsaatmnmcskchkemitkqeqaqlaaspidsivnggdggkgpviaasvn- vavpqveqk tivvqpmlvaetseaaavipkakegpdrcaacrkrvgltgfscrcgnmycsvhrysdkhdcqfdyrtaardaia- kanpvvraekldki cDNA sequence of LUL61567101 from linseed (SEQ ID NO: 25): atggctccttcaccttgcgtccacggctgcacggccaattgcccccgctgccactcttacggacaccccatctt- cgggaactcagatctc gccgctggcggcagcgatacgtccacgtcggtgtttggaaaagtaggatccgtcgtgattcagtcgcctgcgaa- gaatcacgcgttcg gccaagcttgtggcccggtttttccctcgagctcctcccctttccgccgcatcaagttcggccccaaagatggc- gaggggaaaggacc gctgaagccgatcgagaagcagccgtcgaagaagcgtccgttctgcttctctcccgacgagacgattgacgcga- cggttcctccgtc caccaaaccgttcggttcgttccgttccgtctgtgtcacggacgccgacgaggccaggttgaaggcgaaccgcg- agttcttcgctccg gtatcccgcaaacgtggcttcgatccgactgacatgaccttcggtaacgccgccgccgctgcggctaatgcgag- ggaggaagcgaa gaagtggtgcggcagttgcaagaagcgcgtggggctgttagggttcaagtgcaggtgtacgaagttcttctgtg- ggaagcatcggtat cctgaggagcatggttgtacgttcgatcatgtggcgttcgggaggcggattatcgagaaacagaatcctgttct- cgagaccgacaagc tggtggacagaatctga The LU61567101 cDNA is translated into the following amino acid sequence (SEQ ID NO: 26): mapspcvhgctancprchsyghpifgnsdlaaggsdtstsvfgkvgsvviqspaknhafgqacgpvfpsssspf- rrikfgpkdgeg kgplkpiekqpskkrpfcfspdetidatvppstkpfgsfrsvcvtdadearlkanreffapvsrkrgfdptdmt- fgnaaaaaanareeak kwcgsckkrvgllgfkcrctkffcgkhrypeehgctfdhvafgrriiekqnpvletdklvdri cDNA sequence of LU61893412 from linseed (SEQ ID NO: 27): atggaccatgacgaggcaggctgccaggctccttccgatcatcctattctgtgcgttaacaattgcggcttctt- cggaagtgctgccacc atgaacatgtgctcaaagtgccacaaggatacgatgctaaaccaagagcaatccaagcttgctgcttcatcggc- agcaagtatcctc aacggatcgtcgatgagcctcggaagggaactcgttattgctgctaagaccaattcggtagaacccaagaccat- ctccgtccaacca tcttctgcttcaagtgctgaagagagtatcgaaatgaagctgccaaaagaagggcccagtaggtgcaacacttg- caacaaacgtgtc ggtttgaccggattcaaatgtcggtgcgagaacatgttctgcgcaaaccatcgctactcggacaagcacaattg- cccctttgattaccg cactgctggccgtgaagctatctcaaaggccaatcctttggtgaaggcggagaagctcgacaaaatctga The LU61893412 cDNA is translated into the following amino acid sequence (SEQ ID NO: 28): mdhdeagcqapsdhpilcvnncgffgsaatmnmcskchkdtmlnqeqsklaassaasllngssmslgreiviaa- ktnsvepktis vqpssassaeeliemklpkegpsrcntcnkrvgltgfkcrcenmfcanhrysdkhncpfdyrtagreaiskanp- lvkaekldki cDNA sequence of OS39781852 from rice (SEQ ID NO: 29): atggcgcagcgcgacaagaaggatcaggagccgacggagctcagggcgccggagatcacgctgtgcgccaacag- ctgcggatt cccgggcaacccggccacgcagaacctctgccagaactgcttcttggcggccacggcgtccacctcgtcgccgt- cttctttgtcgtcac cggtgctcgacaagcagccgccgaggccggcggcgccgctggttgagcctcaggctcctctcccaccgcctgtg- gaggagatggc ctccgcgctcgcgacggcgccggcgccggtcgccaagacgtcggcggtgaaccggtgctccaggtgccggaagc- gtgtcggcctc accgggttccggtgccggtgcggccacctgttctgcggcgagcaccggtactccgaccgccacggctgcagcta- cgactacaagtc ggcggcgagggacgccatcgccagggacaacccggtggtgcgcgcggccaagatcgttaggttctga The OS39781852 cDNA is translated into the following amino acid sequence (SEQ ID NO: 30): maqrdkkdqeptelrapeitlcanscgfpgnpatqnlcqncflaatastsspsslsspvldkqpprpaaplvep- qaplpppveemas alatapapvaktsavnrcsrcrkrvgltgfrcrcghlfcgehrysdrhgcsydyksaardaiardnpvvraaki- vrf cDNA sequence of OS34701560 from rice (SEQ ID NO: 31): atggccgaagaacaccgatgccaagctcccgaaggtcacagactctgctccaacaactgcggtttctttggtag- ccccgccaccatg aatctctgttccaaatgctacagagacatccgtttgaaggaagaagaacaagccaaaaccaaatccacaatcga- aaccgctctttca ggatcttcctccggccaccgtcaccgcaaccgccgtcgttgcctcctccgtggaatccccttcggcgccggttg- aatccctccctcaacca ccggtgctgatttcgccggatatagccgcaccggttcaggcgaaccggtgcggcgcgtgtaggaagcgcgtggg- gttgacagggttc aagtgcaggtgcggaacaacgttttgtgggagccacaggtaccccgagaaacacgcgtgtggcttcgatttcaa- ggcggtggggag agaggagatagcacgggcgaatcccgtgatcaaaggcgagaagctacggaggatttaa The OS34701560 cDNA is translated into the following amino acid sequence (SEQ ID NO: 32): maeehrcqapeghrlcsnncgffgspatmnlcskcyrdirlkeeeqaktkstietalsgsssatvtatavvass- vespsapveslpqp pvlispdiaapvqanrcgacrkrvgltgfkcrcgttfcgshrypekhacgfdfkavgreelaranpvikgeklr- ri cDNA sequence of OS36821256 from rice (SEQ ID NO: 33): atggcgcagagggagaagaaggtggaggagccgacggagctgagggcgccggagatgacgctctgcgccaacag- ctgcgggt tcccgggcaacccggcgaccaacaacctctgccagaactgcttcttggctgcctcggcgtcttcttcttcttct- tccgccgctgcctcgcc gtcgacgacgtcgttgccggtgtttccggtggtggagaagccgaggcaggccgtacagtcgtcggcggcggcgg- cggtggcgctgg tggttgagcggccgacggcggggccggtggagtcgtcgtcgaaggcgtcgaggtcgtcgtcggtcaaccgatgc- cacagctgccg gaggcgggtgggcctgaccgggttccggtgccgctgcggcgagctctactgcggcgcgcaccggtactccgacc- gccacgactgc agcttcgactacaagtcggcggcgagggacgccatcgccagggagaaccccgtcgtccgcgccgccaagatcgt- taggttctaa The OS36821256 cDNA is translated into the following amino acid sequence (SEQ ID NO: 34): maqrekkveeptelrapemtlcanscgfpgnpatnnlcqncflaasassssssaaaspsttslpvfpvvekprq- avqssaaaaval vverptagpvessskasrsssvnrchscrrrvgltgfrcrcgelycgahrysdrhdcsfdyksaardaiarenp- vvraakivrf cDNA sequence of GM51659494 from soybean (SEQ ID NO: 35): atggctcagaaaaccgagaaagaagaaaccgacttcaaagttccggaaacgattacgctttgcgtcaacaactg- cggcgtcaccg gaaaccctgccacgaataacatgtgccagaagtgcttcactgcctctaccgccaccacttccggcgccggaggt- gccggaatagctt ctccggcgaccagatccggcgtctccgcgcgtcctcagaagagatcttttcctgaagagccctcgccggtggcg- gatcctccttcttcg gaccagacgacgccgtcggaggcgaagcgcgtggtcaaccgctgctccggatgccggcggaaggtcggactcac- cggattccgg tgccggtgcggcgagctcttctgcgccgagcaccggtactccgaccgccacgactgcagctatgactacaaagc- cgccggaagag aagccatcgcgagggagaatccggtgatcagagctgcgaagatcgtcaaagtctga The GM51659494 cDNA is translated into the following amino acid sequence (SEQ ID NO: 36): maqktekeetdfkvpetitlcvnncgvtgnpatnnmcqkcftastattsgaggagiaspatrsgvsarpqkrsf- peepspvadppss dqttpseakrvvnrcsgcrrkvgltgfrcrcgelfcaehrysdrhdcsydykaagreaiarenpviraakivkv cDNA sequence of GM49780101 from soybean (SEQ ID NO: 37): atggagcctcatgatgagactggatgccaggctcctgaacgccccattctttgcattaataattgtggcttctt- tggaagagcagctacca tgaacatgtgttccaagtgttacaaggacatgctgttgaagcaggagcaggacaaatttgcagcatcatccgtt- gaaaacattgtgaat ggcagttccaatggcaatggaaagcaggctgtggctactggtgctgttgctgtacaagttgaagctgtggaggt- caagattgtctgtgct cagagttctgtggattcgtcctccggtgatagtttggagatgaaagccaagactggtcccagtagatgtgctac- atgccggaaacgtgtt ggtttaactggtttcagctgcaaatgtggcaacctcttctgtgcaatgcatcgctattctgataaacatgattg- cccttttgattataggactgt tggtcaggatgccatagctaaagccaaccccataattaaggcagataagctcgacaaaatctag The GM49780101 cDNA is translated into the following amino acid sequence (SEQ ID NO: 38): mephdetgcqaperpilcinncgffgraatmnmcskcykdmllkqeqdkfaassvenlvngssngngkqavatg- avavqveave vkivcaqssvdsssgdslemkaktgpsrcatcrkrvgitgfsckcgnlfcamhrysdkhdcpfdrtvgqdalak- anplikadkldki cDNA sequence of GM59637305 from soybean (SEQ ID NO: 39): atggaccatgacaagactgggtgccaagctcctcctgaaggtcctatattgtgcatcaacaactgtgggttttt- tggaagtgcagctacc atgaacatgtgttctaaatgccacaaagacatattgctgaaacaggagcaggccaagcttgcagcatcatccat- tgggaatattatga atgggtcatcaagcagcactgaaaaggaacctgttgttgctgctgctgctaatattgatatcccagttattcca- gtagagcctaaaactgt ctctgtgcaacctttatttggttcaggtccagaggggagtgttgaggcaaagccgaaggatggaccaaaacgtt- gcagcagctgcaa caagcgagttggtttgacagggtttaattgtcgatgtggtgacctttttttgtgctgtacatcgctactcgaca- agcataattgcccatttgatt accgcactgccgctcaagatgctatagctaaagcaaacccagttgtcaaggctgaaaagcttgataagatctaa The GM59637305 cDNA is translated into the following amino acid sequence (SEQ ID NO: 40): mdhdktgcqappegpilcinncgffgsaatmnmcskchkdillkqeqaklaassignimngsssstekepvvaa- aanldlpvipve pktvsvqplfgsgpegsveakpkdgpkrcsscnkrvgltgfncrcgdlflcctslldkhncpfdyrtaaqdaia- kanpvvkaekldki cDNA sequence of TA55974113 from wheat (SEQ ID NO: 41): atggcgcagcgggatcacaagcaggaggagcccacggagctgcgggcgccggagatcacgctctgcgccaacag- ctgcggctt cccgggcaacccggccacgcagaacctctgccagaactgcttcttggccggcccggcgtccacgtcgccgtctt- cctcctcctcctcct cctcttctctgccgggcgtgtccgcgccgacccccgtcatcgacaggccgaggccggcgccgttggaggcggag- ctggcacgcccc gccgtcgaccttgctccggcgacggaggcgaagccggcgaggacgtcggtgaaccggtgctccagctgccggaa- gcgcgtgggg ctgacggggttccggtgccggtgcggcgacatgttctgcggcgagcaccggtactcggaccggcacgggtggca- gctacgactacaa ggccgccgccagggacgccatcgccagggacaaccccgtcgtgcgcgccgccaagatcgtcaggttctga The TA55974113 cDNA is translated into the following amino acid sequence (SEQ ID NO: 42): maqrdhkqeeptelrapietlcanscgfpgnpalqnlcqncflagpastspssssssssslpgvsaptpvidrp-
rpapleaelarpav dlapateakpartsvnrcsscrkrvgltgfrcrcgdmfcgehrysdrhgcsydykaaardaiardnpvvraaki- vrf The EST65 amino acid sequence (SEQ ID NO: 43): mvaesvlvcrssvvgaglqsfygegakresagpgrsvflgaqvqkmgagmsarsdvrpaavpkasgdvseqtdy- ktfsdeewk krlsqqqfyvarkkgterpftgeywntktagtylcvccktplfssktkfdsgtgwpsyydtigdnvkshmdwsi- pfmprtevvcavcda hlghvfddgprptgkrycinsaaidlkaekqeern The EST12 amino acid sequence (SEQ ID NO: 44): mvvpslpafggqnamlrrnidnntdtlisllqgscsprvsmqqvprsseslenmmgacgqklpyfssfdgpsve- eqedvdegidef ahhvekkrrlsleqvrslernfevenkleperkmqlakelglrprqvavwfqnrrarwktkqlehdyetlkkay- drlkadfeavtldtnalk aevsrlkglsnddvkpaefvqgkcdttshpaspaqsersdlvssrnrttptihvdpvapeeagahltmssdsns- sevmdadsprtsh tsasrstistsvvqpdeglgvaqyphfspenfvgpnmpeicadqslasqvkleeihsfnpdqtflllpnwwdwa The EST307 amino acid sequence (SEQ ID NO: 45): matervsqettsqapegpvmcknlcgffgsqatmglcskcyretvmqakmtalaeqatqaaqatsataaavqpp- apvhetkltce vertmivphqsssyqqdlvtpaaaapqavkssiaapsrpepnrcgscrkrvgltgfkcrcgnlycalhrysdkh- tctydykaagqeai akanplvvaekvvkf
Sequence CWU
1
1
451615DNABrassica napusCDS(1)..(615)methionine sulfoxide reductase family
protein (BN51364980) 1atg gct tct tct agt tgt ttc acc att cag tca
cgt ttc gtc tca gcg 48Met Ala Ser Ser Ser Cys Phe Thr Ile Gln Ser
Arg Phe Val Ser Ala 1 5 10
15 aga aca aag ctc gat tca atc tcc aaa ccg agt ctc
tcc gga ttc gct 96Arg Thr Lys Leu Asp Ser Ile Ser Lys Pro Ser Leu
Ser Gly Phe Ala 20 25
30 tgt cgt tct ctt aca aaa ccc aga aac ttg aat ctc tct
gtt ctt ctt 144Cys Arg Ser Leu Thr Lys Pro Arg Asn Leu Asn Leu Ser
Val Leu Leu 35 40 45
cgg tgt tcc atg ggt tcc ttt aac tct tct cag aaa tca gac
aac gtc 192Arg Cys Ser Met Gly Ser Phe Asn Ser Ser Gln Lys Ser Asp
Asn Val 50 55 60
caa gaa gct gca aag agt gac ttt gct tca ata agt gaa ggt gag
tgg 240Gln Glu Ala Ala Lys Ser Asp Phe Ala Ser Ile Ser Glu Gly Glu
Trp 65 70 75
80 aag aaa cgg cta aca cca gaa cag tat tac atc acc aga cag aag
gga 288Lys Lys Arg Leu Thr Pro Glu Gln Tyr Tyr Ile Thr Arg Gln Lys
Gly 85 90 95
aca gag aga gct ttc act ggt gag tat tgg aat aca aag acc cca gga
336Thr Glu Arg Ala Phe Thr Gly Glu Tyr Trp Asn Thr Lys Thr Pro Gly
100 105 110
gta tac aaa tgt atc tgt tgc gac acg cca ctg ttt gac tca tca aca
384Val Tyr Lys Cys Ile Cys Cys Asp Thr Pro Leu Phe Asp Ser Ser Thr
115 120 125
aag ctt gat agt gga acc ggg tgg cca tcg tat tac caa cct att gga
432Lys Leu Asp Ser Gly Thr Gly Trp Pro Ser Tyr Tyr Gln Pro Ile Gly
130 135 140
aac aat gtg aag tca aag ctg gac ctc tct atc atc ttc atg cct aga
480Asn Asn Val Lys Ser Lys Leu Asp Leu Ser Ile Ile Phe Met Pro Arg
145 150 155 160
caa gaa gtt atc tgt gct gtt tgt aac gcc cat ctt ggt cat gtc ttc
528Gln Glu Val Ile Cys Ala Val Cys Asn Ala His Leu Gly His Val Phe
165 170 175
gat gac ggt cca cga cca acc gga aaa cga tat tgc ctc aac agt gct
576Asp Asp Gly Pro Arg Pro Thr Gly Lys Arg Tyr Cys Leu Asn Ser Ala
180 185 190
gct ctg aaa ctt gag tca ttg gag aga aca aga gaa tga
615Ala Leu Lys Leu Glu Ser Leu Glu Arg Thr Arg Glu
195 200
2204PRTBrassica napus 2Met Ala Ser Ser Ser Cys Phe Thr Ile Gln Ser Arg
Phe Val Ser Ala 1 5 10
15 Arg Thr Lys Leu Asp Ser Ile Ser Lys Pro Ser Leu Ser Gly Phe Ala
20 25 30 Cys Arg Ser
Leu Thr Lys Pro Arg Asn Leu Asn Leu Ser Val Leu Leu 35
40 45 Arg Cys Ser Met Gly Ser Phe Asn
Ser Ser Gln Lys Ser Asp Asn Val 50 55
60 Gln Glu Ala Ala Lys Ser Asp Phe Ala Ser Ile Ser Glu
Gly Glu Trp 65 70 75
80 Lys Lys Arg Leu Thr Pro Glu Gln Tyr Tyr Ile Thr Arg Gln Lys Gly
85 90 95 Thr Glu Arg Ala
Phe Thr Gly Glu Tyr Trp Asn Thr Lys Thr Pro Gly 100
105 110 Val Tyr Lys Cys Ile Cys Cys Asp Thr
Pro Leu Phe Asp Ser Ser Thr 115 120
125 Lys Leu Asp Ser Gly Thr Gly Trp Pro Ser Tyr Tyr Gln Pro
Ile Gly 130 135 140
Asn Asn Val Lys Ser Lys Leu Asp Leu Ser Ile Ile Phe Met Pro Arg 145
150 155 160 Gln Glu Val Ile Cys
Ala Val Cys Asn Ala His Leu Gly His Val Phe 165
170 175 Asp Asp Gly Pro Arg Pro Thr Gly Lys Arg
Tyr Cys Leu Asn Ser Ala 180 185
190 Ala Leu Lys Leu Glu Ser Leu Glu Arg Thr Arg Glu 195
200 3615DNAOryza
sativaCDS(1)..(615)methionine sulfoxide reductase family protein
(OS34096188) 3atg ggc ttc aat att ctg aga acc act tcc atc tcc act cct atc
tct 48Met Gly Phe Asn Ile Leu Arg Thr Thr Ser Ile Ser Thr Pro Ile
Ser 1 5 10 15
tcc tcc aaa tcc aaa ccc att ttc tca act ctt ctt cgt tct tct cct
96Ser Ser Lys Ser Lys Pro Ile Phe Ser Thr Leu Leu Arg Ser Ser Pro
20 25 30
tcc acc att ttc ccc cca aag tcc gtt act ccc acc act ctt ttc gtt
144Ser Thr Ile Phe Pro Pro Lys Ser Val Thr Pro Thr Thr Leu Phe Val
35 40 45
tct gcc acc ccc ttc ttc act ctc cat ccc aag ctt ggt ttt cgt ggt
192Ser Ala Thr Pro Phe Phe Thr Leu His Pro Lys Leu Gly Phe Arg Gly
50 55 60
ggg att gtg gcc atg gcc gca cct ggc tct ctc cgc aaa tcc gag gaa
240Gly Ile Val Ala Met Ala Ala Pro Gly Ser Leu Arg Lys Ser Glu Glu
65 70 75 80
gag tgg cgc gca att ctc tcc cct gaa cag ttt cgg atc ctc agg caa
288Glu Trp Arg Ala Ile Leu Ser Pro Glu Gln Phe Arg Ile Leu Arg Gln
85 90 95
aag ggc acc gag ttc cct gga aca gga gag tat gac aag ttc tat gaa
336Lys Gly Thr Glu Phe Pro Gly Thr Gly Glu Tyr Asp Lys Phe Tyr Glu
100 105 110
gag gga gtt tac aac tgt gct ggt tgt ggg act cca ctc tac agg tcc
384Glu Gly Val Tyr Asn Cys Ala Gly Cys Gly Thr Pro Leu Tyr Arg Ser
115 120 125
ata aca aaa ttc aat tct ggt tgt ggc tgg cca gcc ttc tat gag ggg
432Ile Thr Lys Phe Asn Ser Gly Cys Gly Trp Pro Ala Phe Tyr Glu Gly
130 135 140
att ccc gga gcc ata aat cgc aat ccg gat cct gat ggg atg agg aca
480Ile Pro Gly Ala Ile Asn Arg Asn Pro Asp Pro Asp Gly Met Arg Thr
145 150 155 160
gaa ata acg tgt gct gct tgt ggg gga cat cta ggt cac gtc ttt aaa
528Glu Ile Thr Cys Ala Ala Cys Gly Gly His Leu Gly His Val Phe Lys
165 170 175
gga gaa gga ttt cca aca ccc act aac gaa cgc cat tgt gtc aat agc
576Gly Glu Gly Phe Pro Thr Pro Thr Asn Glu Arg His Cys Val Asn Ser
180 185 190
att tcg ctg aaa ttt gcg cca gcc aat tct tat tct taa
615Ile Ser Leu Lys Phe Ala Pro Ala Asn Ser Tyr Ser
195 200
4204PRTOryza sativa 4Met Gly Phe Asn Ile Leu Arg Thr Thr Ser Ile Ser Thr
Pro Ile Ser 1 5 10 15
Ser Ser Lys Ser Lys Pro Ile Phe Ser Thr Leu Leu Arg Ser Ser Pro
20 25 30 Ser Thr Ile Phe
Pro Pro Lys Ser Val Thr Pro Thr Thr Leu Phe Val 35
40 45 Ser Ala Thr Pro Phe Phe Thr Leu His
Pro Lys Leu Gly Phe Arg Gly 50 55
60 Gly Ile Val Ala Met Ala Ala Pro Gly Ser Leu Arg Lys
Ser Glu Glu 65 70 75
80 Glu Trp Arg Ala Ile Leu Ser Pro Glu Gln Phe Arg Ile Leu Arg Gln
85 90 95 Lys Gly Thr Glu
Phe Pro Gly Thr Gly Glu Tyr Asp Lys Phe Tyr Glu 100
105 110 Glu Gly Val Tyr Asn Cys Ala Gly Cys
Gly Thr Pro Leu Tyr Arg Ser 115 120
125 Ile Thr Lys Phe Asn Ser Gly Cys Gly Trp Pro Ala Phe Tyr
Glu Gly 130 135 140
Ile Pro Gly Ala Ile Asn Arg Asn Pro Asp Pro Asp Gly Met Arg Thr 145
150 155 160 Glu Ile Thr Cys Ala
Ala Cys Gly Gly His Leu Gly His Val Phe Lys 165
170 175 Gly Glu Gly Phe Pro Thr Pro Thr Asn Glu
Arg His Cys Val Asn Ser 180 185
190 Ile Ser Leu Lys Phe Ala Pro Ala Asn Ser Tyr Ser 195
200 5645DNAOryza
sativaCDS(1)..(645)methionine sulfoxide reductase family protein
(OS32583643) 5atg gcc atg cgg caa tac gcg gct gct acc gct gcc tcc tcc agt
ttc 48Met Ala Met Arg Gln Tyr Ala Ala Ala Thr Ala Ala Ser Ser Ser
Phe 1 5 10 15
aga gca cgt cca cgg gcg cgc ccc tcc tgc ctc cca gcc gcc gcc ctg
96Arg Ala Arg Pro Arg Ala Arg Pro Ser Cys Leu Pro Ala Ala Ala Leu
20 25 30
ccc ttg gcg cct tgc tgt ggt gtg gcg tgg agc cgt gct agc tac agg
144Pro Leu Ala Pro Cys Cys Gly Val Ala Trp Ser Arg Ala Ser Tyr Arg
35 40 45
cga gcc tcc gtt cgt gcc atg ggt gcc gct tca tcg tct tcg tcg tcg
192Arg Ala Ser Val Arg Ala Met Gly Ala Ala Ser Ser Ser Ser Ser Ser
50 55 60
tcg tcg tcg tct ccg tcg ccg cag ggt caa gcc caa gcc caa gcc caa
240Ser Ser Ser Ser Pro Ser Pro Gln Gly Gln Ala Gln Ala Gln Ala Gln
65 70 75 80
ggt aaa ccg aac tac agt aca tct ctg act gat gag gag tgg agg aag
288Gly Lys Pro Asn Tyr Ser Thr Ser Leu Thr Asp Glu Glu Trp Arg Lys
85 90 95
cgc ctg aca aaa gat cag tat tac att act cgg cag aag ggc aca gaa
336Arg Leu Thr Lys Asp Gln Tyr Tyr Ile Thr Arg Gln Lys Gly Thr Glu
100 105 110
aga gca ttt act ggg gaa tac tgg aac acc aaa acc ccg ggc atc tac
384Arg Ala Phe Thr Gly Glu Tyr Trp Asn Thr Lys Thr Pro Gly Ile Tyr
115 120 125 cat
tgt gtc tgc tgt gac acc cct ctt ttt gag tca tcg acc aaa ttt 432His
Cys Val Cys Cys Asp Thr Pro Leu Phe Glu Ser Ser Thr Lys Phe
130 135 140 gat
agt ggt act ggg tgg ccg tca tat tat caa ccc att gga gat aat 480Asp
Ser Gly Thr Gly Trp Pro Ser Tyr Tyr Gln Pro Ile Gly Asp Asn 145
150 155 160 gta aag
tgc aag ctt gat atg tcc atc ata ttc atg cct cgg act gag 528Val Lys
Cys Lys Leu Asp Met Ser Ile Ile Phe Met Pro Arg Thr Glu
165 170 175 gtg ctg tgt
gct gtc tgt gac gct cat ctg ggg cac gtg ttt gat gat 576Val Leu Cys
Ala Val Cys Asp Ala His Leu Gly His Val Phe Asp Asp
180 185 190 ggg cca cga
cca aca ggg aaa aga tac tgt atc aat agc gca tct ctc 624Gly Pro Arg
Pro Thr Gly Lys Arg Tyr Cys Ile Asn Ser Ala Ser Leu 195
200 205 aag ctg aag aag
acc cag tag 645Lys Leu Lys Lys
Thr Gln 210
6214PRTOryza sativa
6Met Ala Met Arg Gln Tyr Ala Ala Ala Thr Ala Ala Ser Ser Ser Phe 1
5 10 15 Arg Ala Arg Pro
Arg Ala Arg Pro Ser Cys Leu Pro Ala Ala Ala Leu 20
25 30 Pro Leu Ala Pro Cys Cys Gly Val Ala
Trp Ser Arg Ala Ser Tyr Arg 35 40
45 Arg Ala Ser Val Arg Ala Met Gly Ala Ala Ser Ser Ser Ser
Ser Ser 50 55 60
Ser Ser Ser Ser Pro Ser Pro Gln Gly Gln Ala Gln Ala Gln Ala Gln 65
70 75 80 Gly Lys Pro Asn Tyr
Ser Thr Ser Leu Thr Asp Glu Glu Trp Arg Lys 85
90 95 Arg Leu Thr Lys Asp Gln Tyr Tyr Ile Thr
Arg Gln Lys Gly Thr Glu 100 105
110 Arg Ala Phe Thr Gly Glu Tyr Trp Asn Thr Lys Thr Pro Gly Ile
Tyr 115 120 125 His
Cys Val Cys Cys Asp Thr Pro Leu Phe Glu Ser Ser Thr Lys Phe 130
135 140 Asp Ser Gly Thr Gly Trp
Pro Ser Tyr Tyr Gln Pro Ile Gly Asp Asn 145 150
155 160 Val Lys Cys Lys Leu Asp Met Ser Ile Ile Phe
Met Pro Arg Thr Glu 165 170
175 Val Leu Cys Ala Val Cys Asp Ala His Leu Gly His Val Phe Asp Asp
180 185 190 Gly Pro
Arg Pro Thr Gly Lys Arg Tyr Cys Ile Asn Ser Ala Ser Leu 195
200 205 Lys Leu Lys Lys Thr Gln
210 7609DNAGlycine maxCDS(1)..(609)methionine sulfoxide
reductase family protein (GM53626178) 7atg gga ttg agt att ctg aga
agc act tcc att tcc act cct atc tct 48Met Gly Leu Ser Ile Leu Arg
Ser Thr Ser Ile Ser Thr Pro Ile Ser 1 5
10 15 tcc tcc aaa tcc aaa ccc att ttc
tca act ctt gtt cgt tca tct ttc 96Ser Ser Lys Ser Lys Pro Ile Phe
Ser Thr Leu Val Arg Ser Ser Phe 20
25 30 gcc tcc att tcc ccc aca aag tgt
gtt act ccc acc act ctt ttc gtt 144Ala Ser Ile Ser Pro Thr Lys Cys
Val Thr Pro Thr Thr Leu Phe Val 35 40
45 tct gcc acc ccc ttc ttc acc gcc tca
ccc aag cgc ggc ttt cgt ggt 192Ser Ala Thr Pro Phe Phe Thr Ala Ser
Pro Lys Arg Gly Phe Arg Gly 50 55
60 ggg att gtg gcc atg gcc gcc gct ggc tcg
ctc cgc aaa tca gag gaa 240Gly Ile Val Ala Met Ala Ala Ala Gly Ser
Leu Arg Lys Ser Glu Glu 65 70
75 80 gag tgg cgc gca gtt ctc tcc cct gaa cag
ttt cgt att ctc agg caa 288Glu Trp Arg Ala Val Leu Ser Pro Glu Gln
Phe Arg Ile Leu Arg Gln 85 90
95 aag ggc acc gag ttc cct gga aca gga gag tat
gac aag ttc ttt gat 336Lys Gly Thr Glu Phe Pro Gly Thr Gly Glu Tyr
Asp Lys Phe Phe Asp 100 105
110 gag gga gtt tac aac tgt gct ggt tgt ggg aca cct
ctc tac agg tcc 384Glu Gly Val Tyr Asn Cys Ala Gly Cys Gly Thr Pro
Leu Tyr Arg Ser 115 120
125 tta aca aaa ttc aat tct ggt tgt ggc tgg cca gcc
ttc tat gag ggg 432Leu Thr Lys Phe Asn Ser Gly Cys Gly Trp Pro Ala
Phe Tyr Glu Gly 130 135 140
att cct gga gcc ata aat cgc aat ccg gac cct gat ggg
atg agg aca 480Ile Pro Gly Ala Ile Asn Arg Asn Pro Asp Pro Asp Gly
Met Arg Thr 145 150 155
160 gaa ata acg tgt gct gct tgt ggg gga cat cta ggt cac gtc
ttt aaa 528Glu Ile Thr Cys Ala Ala Cys Gly Gly His Leu Gly His Val
Phe Lys 165 170
175 gga gaa gga ttt cca acg ccc act aac gaa cgc cat tgt gtc
aat agc 576Gly Glu Gly Phe Pro Thr Pro Thr Asn Glu Arg His Cys Val
Asn Ser 180 185 190
att tca ctg aaa ttt gcg cca gcc aat tct taa
609Ile Ser Leu Lys Phe Ala Pro Ala Asn Ser
195 200
8202PRTGlycine max 8Met Gly Leu Ser Ile Leu Arg Ser Thr Ser Ile
Ser Thr Pro Ile Ser 1 5 10
15 Ser Ser Lys Ser Lys Pro Ile Phe Ser Thr Leu Val Arg Ser Ser Phe
20 25 30 Ala Ser
Ile Ser Pro Thr Lys Cys Val Thr Pro Thr Thr Leu Phe Val 35
40 45 Ser Ala Thr Pro Phe Phe Thr
Ala Ser Pro Lys Arg Gly Phe Arg Gly 50 55
60 Gly Ile Val Ala Met Ala Ala Ala Gly Ser Leu Arg
Lys Ser Glu Glu 65 70 75
80 Glu Trp Arg Ala Val Leu Ser Pro Glu Gln Phe Arg Ile Leu Arg Gln
85 90 95 Lys Gly Thr
Glu Phe Pro Gly Thr Gly Glu Tyr Asp Lys Phe Phe Asp 100
105 110 Glu Gly Val Tyr Asn Cys Ala Gly
Cys Gly Thr Pro Leu Tyr Arg Ser 115 120
125 Leu Thr Lys Phe Asn Ser Gly Cys Gly Trp Pro Ala Phe
Tyr Glu Gly 130 135 140
Ile Pro Gly Ala Ile Asn Arg Asn Pro Asp Pro Asp Gly Met Arg Thr 145
150 155 160 Glu Ile Thr Cys
Ala Ala Cys Gly Gly His Leu Gly His Val Phe Lys 165
170 175 Gly Glu Gly Phe Pro Thr Pro Thr Asn
Glu Arg His Cys Val Asn Ser 180 185
190 Ile Ser Leu Lys Phe Ala Pro Ala Asn Ser 195
200 9621DNATriticum aestivumCDS(1)..(621)methionine
sulfoxide reductase family protein (TA6540264) 9atg gcg tcg ccc cac
gcc cac ccg gcc acg cgg ccc ctc tca tcg ctc 48Met Ala Ser Pro His
Ala His Pro Ala Thr Arg Pro Leu Ser Ser Leu 1 5
10 15 ccg tcc ctc ctc ctc gcc
cgc tcc tcc tcc gcc gcc acc gcc gcc gcg 96Pro Ser Leu Leu Leu Ala
Arg Ser Ser Ser Ala Ala Thr Ala Ala Ala 20
25 30 tcg tcc gcc cgc ccc gcc tcc
ctc tcc ctc tcg tgc tcg cgg tcg cgg 144Ser Ser Ala Arg Pro Ala Ser
Leu Ser Leu Ser Cys Ser Arg Ser Arg 35
40 45 gcg cgg gcc tac tgc cca gcc
gga cga cgg ttg ccg ggc gcc gtg gtg 192Ala Arg Ala Tyr Cys Pro Ala
Gly Arg Arg Leu Pro Gly Ala Val Val 50 55
60 gct atg tcg tcg gcg gcg ccc acg
ccg ggg ccc gtg cag aag tcg gag 240Ala Met Ser Ser Ala Ala Pro Thr
Pro Gly Pro Val Gln Lys Ser Glu 65 70
75 80 gag gag tgg gag gcc gtc ctc acg ccg
gag cag ttc cgc atc ctc cgc 288Glu Glu Trp Glu Ala Val Leu Thr Pro
Glu Gln Phe Arg Ile Leu Arg 85
90 95 cgc aag ggc acc gag tat cct gga aca
ggt gaa tat gac aag ttc ttc 336Arg Lys Gly Thr Glu Tyr Pro Gly Thr
Gly Glu Tyr Asp Lys Phe Phe 100 105
110 agt gag ggt att tac gga tgt gct ggc tgt
gga acc ccc ttg tac aaa 384Ser Glu Gly Ile Tyr Gly Cys Ala Gly Cys
Gly Thr Pro Leu Tyr Lys 115 120
125 tca tct acg aag ttc aac tca ggg tgt ggt tgg
cca gca ttc tat gaa 432Ser Ser Thr Lys Phe Asn Ser Gly Cys Gly Trp
Pro Ala Phe Tyr Glu 130 135
140 gga ttt cct gga gcc ata aaa cgg acg gcg gat
cct gat ggg agg cga 480Gly Phe Pro Gly Ala Ile Lys Arg Thr Ala Asp
Pro Asp Gly Arg Arg 145 150 155
160 att gag atc aca tgt gct gct tgt gaa gga cat ctg
ggg cat gtg ttc 528Ile Glu Ile Thr Cys Ala Ala Cys Glu Gly His Leu
Gly His Val Phe 165 170
175 aaa ggg gag ggg ttc aac acg ccg act gat gag cga cac
tgc gtc aac 576Lys Gly Glu Gly Phe Asn Thr Pro Thr Asp Glu Arg His
Cys Val Asn 180 185
190 agt atc tca ctc aag ttc gtt ccg gcc tct gaa gag gct
agt tga 621Ser Ile Ser Leu Lys Phe Val Pro Ala Ser Glu Glu Ala
Ser 195 200 205
10206PRTTriticum aestivum 10Met Ala Ser Pro His Ala His
Pro Ala Thr Arg Pro Leu Ser Ser Leu 1 5
10 15 Pro Ser Leu Leu Leu Ala Arg Ser Ser Ser Ala
Ala Thr Ala Ala Ala 20 25
30 Ser Ser Ala Arg Pro Ala Ser Leu Ser Leu Ser Cys Ser Arg Ser
Arg 35 40 45 Ala
Arg Ala Tyr Cys Pro Ala Gly Arg Arg Leu Pro Gly Ala Val Val 50
55 60 Ala Met Ser Ser Ala Ala
Pro Thr Pro Gly Pro Val Gln Lys Ser Glu 65 70
75 80 Glu Glu Trp Glu Ala Val Leu Thr Pro Glu Gln
Phe Arg Ile Leu Arg 85 90
95 Arg Lys Gly Thr Glu Tyr Pro Gly Thr Gly Glu Tyr Asp Lys Phe Phe
100 105 110 Ser Glu
Gly Ile Tyr Gly Cys Ala Gly Cys Gly Thr Pro Leu Tyr Lys 115
120 125 Ser Ser Thr Lys Phe Asn Ser
Gly Cys Gly Trp Pro Ala Phe Tyr Glu 130 135
140 Gly Phe Pro Gly Ala Ile Lys Arg Thr Ala Asp Pro
Asp Gly Arg Arg 145 150 155
160 Ile Glu Ile Thr Cys Ala Ala Cys Glu Gly His Leu Gly His Val Phe
165 170 175 Lys Gly Glu
Gly Phe Asn Thr Pro Thr Asp Glu Arg His Cys Val Asn 180
185 190 Ser Ile Ser Leu Lys Phe Val Pro
Ala Ser Glu Glu Ala Ser 195 200
205 11933DNABrassica napusCDS(1)..(933)homeodomain leucine zipper
protein (BN45206322) 11atg atg aag aga tta agc agt tca gat tca gtg
ggt ggt ctc atc tct 48Met Met Lys Arg Leu Ser Ser Ser Asp Ser Val
Gly Gly Leu Ile Ser 1 5 10
15 tta tgt ccc act act tcc aca gat cag ccg aat cca
aga aga tgc ggg 96Leu Cys Pro Thr Thr Ser Thr Asp Gln Pro Asn Pro
Arg Arg Cys Gly 20 25
30 aga gaa ttt cag tcg atg ctc gaa ggt tac gag gag gaa
gaa gaa gaa 144Arg Glu Phe Gln Ser Met Leu Glu Gly Tyr Glu Glu Glu
Glu Glu Glu 35 40 45
gcc ata acc gag gaa aga gga caa acc ggt tta gcc gag aag
aag aga 192Ala Ile Thr Glu Glu Arg Gly Gln Thr Gly Leu Ala Glu Lys
Lys Arg 50 55 60
cgg tta aac att aac caa gtt aaa gcc ttg gag aaa aat ttc gag
tta 240Arg Leu Asn Ile Asn Gln Val Lys Ala Leu Glu Lys Asn Phe Glu
Leu 65 70 75
80 gag aac aag ctt gag cct gag agg aaa gtg aag tta gct caa gaa
ctt 288Glu Asn Lys Leu Glu Pro Glu Arg Lys Val Lys Leu Ala Gln Glu
Leu 85 90 95
ggt ctc caa cct cgt caa gta gct gtt tgg ttt cag aac cgc cgt gcg
336Gly Leu Gln Pro Arg Gln Val Ala Val Trp Phe Gln Asn Arg Arg Ala
100 105 110
cgg tgg aag aca aaa cag ctt gag aaa gat tac ggt gtt ctc aaa acg
384Arg Trp Lys Thr Lys Gln Leu Glu Lys Asp Tyr Gly Val Leu Lys Thr
115 120 125
caa tac gat tct ctc cgc cat aac ttt gat tcc ctc cgc cgt gaa aat
432Gln Tyr Asp Ser Leu Arg His Asn Phe Asp Ser Leu Arg Arg Glu Asn
130 135 140
gaa tct ctt ctt caa gag atc ggt aaa cta aaa gct aag ctt aac gga
480Glu Ser Leu Leu Gln Glu Ile Gly Lys Leu Lys Ala Lys Leu Asn Gly
145 150 155 160
gaa gaa gaa gga gat gat gtt gat gaa gaa gag aac aac ttg gcg acg
528Glu Glu Glu Gly Asp Asp Val Asp Glu Glu Glu Asn Asn Leu Ala Thr
165 170 175
atg gag agt gat gtt tcc gtc aag gaa gaa gaa gtt tcg ttg ccg gag
576Met Glu Ser Asp Val Ser Val Lys Glu Glu Glu Val Ser Leu Pro Glu
180 185 190
cag atc aca gag ccg ccg tct tct cct ccg cag ctt cta gag cat tcc
624Gln Ile Thr Glu Pro Pro Ser Ser Pro Pro Gln Leu Leu Glu His Ser
195 200 205
gac agt ttc aat tac cgg agt ttc acc gac ctc cgc gac ctt ctt ccg
672Asp Ser Phe Asn Tyr Arg Ser Phe Thr Asp Leu Arg Asp Leu Leu Pro
210 215 220
tta aag gcc gcg gct tcc tcc gtc gcc gcc gct gga tcg tcg gac agt
720Leu Lys Ala Ala Ala Ser Ser Val Ala Ala Ala Gly Ser Ser Asp Ser
225 230 235 240
agc gat tcg agc gcc gtg ttg aac gag gaa agt agc tct aac gtt acg
768Ser Asp Ser Ser Ala Val Leu Asn Glu Glu Ser Ser Ser Asn Val Thr
245 250 255
gcg gct ccg gcg acg gtt ccc ggc ggc agt ttc ttg cag ttt gtg aaa
816Ala Ala Pro Ala Thr Val Pro Gly Gly Ser Phe Leu Gln Phe Val Lys
260 265 270
atg gag cag acg gag gat cac gac gac ttt ctg agt gga gaa gaa gcg
864Met Glu Gln Thr Glu Asp His Asp Asp Phe Leu Ser Gly Glu Glu Ala
275 280 285
tgc ggg ttt ttc tcc gat gaa cag cca ccg tct ctg cac tgg tat tcc
912Cys Gly Phe Phe Ser Asp Glu Gln Pro Pro Ser Leu His Trp Tyr Ser
290 295 300
acc gtt gat cag tgg aac tga
933Thr Val Asp Gln Trp Asn
305 310
12310PRTBrassica napus 12Met Met Lys Arg Leu Ser Ser Ser Asp Ser Val Gly
Gly Leu Ile Ser 1 5 10
15 Leu Cys Pro Thr Thr Ser Thr Asp Gln Pro Asn Pro Arg Arg Cys Gly
20 25 30 Arg Glu Phe
Gln Ser Met Leu Glu Gly Tyr Glu Glu Glu Glu Glu Glu 35
40 45 Ala Ile Thr Glu Glu Arg Gly Gln
Thr Gly Leu Ala Glu Lys Lys Arg 50 55
60 Arg Leu Asn Ile Asn Gln Val Lys Ala Leu Glu Lys Asn
Phe Glu Leu 65 70 75
80 Glu Asn Lys Leu Glu Pro Glu Arg Lys Val Lys Leu Ala Gln Glu Leu
85 90 95 Gly Leu Gln Pro
Arg Gln Val Ala Val Trp Phe Gln Asn Arg Arg Ala 100
105 110 Arg Trp Lys Thr Lys Gln Leu Glu Lys
Asp Tyr Gly Val Leu Lys Thr 115 120
125 Gln Tyr Asp Ser Leu Arg His Asn Phe Asp Ser Leu Arg Arg
Glu Asn 130 135 140
Glu Ser Leu Leu Gln Glu Ile Gly Lys Leu Lys Ala Lys Leu Asn Gly 145
150 155 160 Glu Glu Glu Gly Asp
Asp Val Asp Glu Glu Glu Asn Asn Leu Ala Thr 165
170 175 Met Glu Ser Asp Val Ser Val Lys Glu Glu
Glu Val Ser Leu Pro Glu 180 185
190 Gln Ile Thr Glu Pro Pro Ser Ser Pro Pro Gln Leu Leu Glu His
Ser 195 200 205 Asp
Ser Phe Asn Tyr Arg Ser Phe Thr Asp Leu Arg Asp Leu Leu Pro 210
215 220 Leu Lys Ala Ala Ala Ser
Ser Val Ala Ala Ala Gly Ser Ser Asp Ser 225 230
235 240 Ser Asp Ser Ser Ala Val Leu Asn Glu Glu Ser
Ser Ser Asn Val Thr 245 250
255 Ala Ala Pro Ala Thr Val Pro Gly Gly Ser Phe Leu Gln Phe Val Lys
260 265 270 Met Glu
Gln Thr Glu Asp His Asp Asp Phe Leu Ser Gly Glu Glu Ala 275
280 285 Cys Gly Phe Phe Ser Asp Glu
Gln Pro Pro Ser Leu His Trp Tyr Ser 290 295
300 Thr Val Asp Gln Trp Asn 305 310
13996DNAGlycine maxCDS(1)..(996)homeodomain leucine zipper protein
(GM48923793) 13atg gcg ggt agt gga agt gcc ttt tcc aac atc act agc ttt
ctt cgc 48Met Ala Gly Ser Gly Ser Ala Phe Ser Asn Ile Thr Ser Phe
Leu Arg 1 5 10
15 acc caa caa ccc tct tct caa cct ctc gat tct tct ctc ttc
ctc tct 96Thr Gln Gln Pro Ser Ser Gln Pro Leu Asp Ser Ser Leu Phe
Leu Ser 20 25 30
gca cct tcc tct gct cct ttc ctc ggt tcg aga tcc atg atg agt
ttt 144Ala Pro Ser Ser Ala Pro Phe Leu Gly Ser Arg Ser Met Met Ser
Phe 35 40 45
gat gga gaa gga ggg aag ggg tgt aac ggc tcc ttc ttc cgc gcg ttt
192Asp Gly Glu Gly Gly Lys Gly Cys Asn Gly Ser Phe Phe Arg Ala Phe
50 55 60
gac atg gac gac aat ggg gat gag tgc atg gac gag tac ttt cat caa
240Asp Met Asp Asp Asn Gly Asp Glu Cys Met Asp Glu Tyr Phe His Gln
65 70 75 80
ccc gag aag aag cga cgt ctc tct gcg agc cag gtt cag ttt cta gag
288Pro Glu Lys Lys Arg Arg Leu Ser Ala Ser Gln Val Gln Phe Leu Glu
85 90 95
aag agc ttc gag gag gag aac aag ctt gaa ccc gag aga aag acc aaa
336Lys Ser Phe Glu Glu Glu Asn Lys Leu Glu Pro Glu Arg Lys Thr Lys
100 105 110
cta gcc aaa gac ctt ggt ttg cag cca cgg caa gtt gct att tgg ttc
384Leu Ala Lys Asp Leu Gly Leu Gln Pro Arg Gln Val Ala Ile Trp Phe
115 120 125
cag aac cgt aga gct cgg tgg aag aac aaa cag ctg gag aag gat tac
432Gln Asn Arg Arg Ala Arg Trp Lys Asn Lys Gln Leu Glu Lys Asp Tyr
130 135 140
gag act ctg cat gca agt ttt gag agt ctc aag tcc aac tat gac tgt
480Glu Thr Leu His Ala Ser Phe Glu Ser Leu Lys Ser Asn Tyr Asp Cys
145 150 155 160
ctt ctc aag gag aaa gac aag tta aaa gct gag gtg gcg agc ctc act
528Leu Leu Lys Glu Lys Asp Lys Leu Lys Ala Glu Val Ala Ser Leu Thr
165 170 175
gag aag gtg ctt gca aga ggg aaa caa gag ggg cac atg aag cag gct
576Glu Lys Val Leu Ala Arg Gly Lys Gln Glu Gly His Met Lys Gln Ala
180 185 190
gaa agt gaa agt gaa gaa aca aaa gga tta ttg cat ttg cag gaa cag
624Glu Ser Glu Ser Glu Glu Thr Lys Gly Leu Leu His Leu Gln Glu Gln
195 200 205
gaa cca ccc cag agg ctt tta ctg caa tca gtt tcg gag gga gaa gga
672Glu Pro Pro Gln Arg Leu Leu Leu Gln Ser Val Ser Glu Gly Glu Gly
210 215 220
tcc aaa gtc tct tct gtc gtt ggg ggt tgt aaa cag gaa gat atc agt
720Ser Lys Val Ser Ser Val Val Gly Gly Cys Lys Gln Glu Asp Ile Ser
225 230 235 240
tca gca agg agt gac att ttg gat tca gat agt cca cat tac acc gat
768Ser Ala Arg Ser Asp Ile Leu Asp Ser Asp Ser Pro His Tyr Thr Asp
245 250 255
gga gtt cac tct gcg ctg cta gag cat ggt gat tct tct tat gtg ttt
816Gly Val His Ser Ala Leu Leu Glu His Gly Asp Ser Ser Tyr Val Phe
260 265 270
gag cct gat caa tca gat atg tca caa gat gaa gaa gat aac ctc agc
864Glu Pro Asp Gln Ser Asp Met Ser Gln Asp Glu Glu Asp Asn Leu Ser
275 280 285
aag agt ctc tac cct tcg tac ctc ttt ccc aaa ctt gaa gaa gat gtg
912Lys Ser Leu Tyr Pro Ser Tyr Leu Phe Pro Lys Leu Glu Glu Asp Val
290 295 300
gat tac tcc gac cca cct gaa agt tct tgt aat ttt gga ttt cct gag
960Asp Tyr Ser Asp Pro Pro Glu Ser Ser Cys Asn Phe Gly Phe Pro Glu
305 310 315 320
gaa gat cat gtc ctt tgg acc tgg gct tac tac taa
996Glu Asp His Val Leu Trp Thr Trp Ala Tyr Tyr
325 330
14331PRTGlycine max 14Met Ala Gly Ser Gly Ser Ala Phe Ser Asn Ile Thr Ser
Phe Leu Arg 1 5 10 15
Thr Gln Gln Pro Ser Ser Gln Pro Leu Asp Ser Ser Leu Phe Leu Ser
20 25 30 Ala Pro Ser Ser
Ala Pro Phe Leu Gly Ser Arg Ser Met Met Ser Phe 35
40 45 Asp Gly Glu Gly Gly Lys Gly Cys Asn
Gly Ser Phe Phe Arg Ala Phe 50 55
60 Asp Met Asp Asp Asn Gly Asp Glu Cys Met Asp Glu Tyr
Phe His Gln 65 70 75
80 Pro Glu Lys Lys Arg Arg Leu Ser Ala Ser Gln Val Gln Phe Leu Glu
85 90 95 Lys Ser Phe Glu
Glu Glu Asn Lys Leu Glu Pro Glu Arg Lys Thr Lys 100
105 110 Leu Ala Lys Asp Leu Gly Leu Gln Pro
Arg Gln Val Ala Ile Trp Phe 115 120
125 Gln Asn Arg Arg Ala Arg Trp Lys Asn Lys Gln Leu Glu Lys
Asp Tyr 130 135 140
Glu Thr Leu His Ala Ser Phe Glu Ser Leu Lys Ser Asn Tyr Asp Cys 145
150 155 160 Leu Leu Lys Glu Lys
Asp Lys Leu Lys Ala Glu Val Ala Ser Leu Thr 165
170 175 Glu Lys Val Leu Ala Arg Gly Lys Gln Glu
Gly His Met Lys Gln Ala 180 185
190 Glu Ser Glu Ser Glu Glu Thr Lys Gly Leu Leu His Leu Gln Glu
Gln 195 200 205 Glu
Pro Pro Gln Arg Leu Leu Leu Gln Ser Val Ser Glu Gly Glu Gly 210
215 220 Ser Lys Val Ser Ser Val
Val Gly Gly Cys Lys Gln Glu Asp Ile Ser 225 230
235 240 Ser Ala Arg Ser Asp Ile Leu Asp Ser Asp Ser
Pro His Tyr Thr Asp 245 250
255 Gly Val His Ser Ala Leu Leu Glu His Gly Asp Ser Ser Tyr Val Phe
260 265 270 Glu Pro
Asp Gln Ser Asp Met Ser Gln Asp Glu Glu Asp Asn Leu Ser 275
280 285 Lys Ser Leu Tyr Pro Ser Tyr
Leu Phe Pro Lys Leu Glu Glu Asp Val 290 295
300 Asp Tyr Ser Asp Pro Pro Glu Ser Ser Cys Asn Phe
Gly Phe Pro Glu 305 310 315
320 Glu Asp His Val Leu Trp Thr Trp Ala Tyr Tyr 325
330 151023DNATriticum aestivumCDS(1)..(1023)homeodomain
leucine zipper protein (TA55969932) 15atg gag ccc ggc cgg ctc atc ttc aac
acg tcg ggc tcc ggc aac gga 48Met Glu Pro Gly Arg Leu Ile Phe Asn
Thr Ser Gly Ser Gly Asn Gly 1 5
10 15 cag atg ctc ttc atg gac tgc ggc gcg
ggc ggc atc gcc ggc gcg gcc 96Gln Met Leu Phe Met Asp Cys Gly Ala
Gly Gly Ile Ala Gly Ala Ala 20 25
30 ggc atg ttc cat cga ggg gtg aga ccg gtc
ctc ggc ggc atg gaa gaa 144Gly Met Phe His Arg Gly Val Arg Pro Val
Leu Gly Gly Met Glu Glu 35 40
45 ggg cgc ggc gtg aag cgg ccc ttc ttc acc tcg
ccg gat gac atg ctg 192Gly Arg Gly Val Lys Arg Pro Phe Phe Thr Ser
Pro Asp Asp Met Leu 50 55
60 gag gag gag tac tac gac gag cag ctc ccg gag
aag aag cgg cgc ctc 240Glu Glu Glu Tyr Tyr Asp Glu Gln Leu Pro Glu
Lys Lys Arg Arg Leu 65 70 75
80 acg ccg gag cag gtc cac ctg ctg gag agg agc ttc
gag gag gag aac 288Thr Pro Glu Gln Val His Leu Leu Glu Arg Ser Phe
Glu Glu Glu Asn 85 90
95 aag ctg gag ccg gag agg aag acg gag ctg gcc cgc aag
ctc ggg ctg 336Lys Leu Glu Pro Glu Arg Lys Thr Glu Leu Ala Arg Lys
Leu Gly Leu 100 105
110 cag cca cgg cag gtg gcc gtc tgg ttc cag aac cgc cgc
gcc cgg tgg 384Gln Pro Arg Gln Val Ala Val Trp Phe Gln Asn Arg Arg
Ala Arg Trp 115 120 125
aag aca aag acg ctg gag cgc gac ttc gac cgc ctc aag gcg
tcc ttc 432Lys Thr Lys Thr Leu Glu Arg Asp Phe Asp Arg Leu Lys Ala
Ser Phe 130 135 140
gac gcc ctc cgc gcc gac cac gac gcg ctc ctc cag gac aac cac
cgg 480Asp Ala Leu Arg Ala Asp His Asp Ala Leu Leu Gln Asp Asn His
Arg 145 150 155
160 ctc cgg tca cag gtg gta acg ttg acc gag aag atg caa gat aag
gag 528Leu Arg Ser Gln Val Val Thr Leu Thr Glu Lys Met Gln Asp Lys
Glu 165 170 175
gcg ccg gaa ggc agc ttc ggt gca gcc gcc gac gcc tcg gag ccg gag
576Ala Pro Glu Gly Ser Phe Gly Ala Ala Ala Asp Ala Ser Glu Pro Glu
180 185 190
cag gcg gcg gcg gag gcg aag gct tcc ttg gcc gac gcc gag gag cag
624Gln Ala Ala Ala Glu Ala Lys Ala Ser Leu Ala Asp Ala Glu Glu Gln
195 200 205
gcc gcg gca gcg gag gcg ttc gag gtg gtg cag cag cag ctg cac gtg
672Ala Ala Ala Ala Glu Ala Phe Glu Val Val Gln Gln Gln Leu His Val
210 215 220
aag gac gag gag agg ctg agc ccg ggg agc ggc ggg agc gcg gtg ctg
720Lys Asp Glu Glu Arg Leu Ser Pro Gly Ser Gly Gly Ser Ala Val Leu
225 230 235 240
gac gcg agg gac gcg ctg ctc ggg agc gga tgc ggc ctc gcc ggc gtg
768Asp Ala Arg Asp Ala Leu Leu Gly Ser Gly Cys Gly Leu Ala Gly Val
245 250 255
gtg gac agc agc gtg gac tcg tac tgc ttc ccg ggg ggc gcc ggc ggc
816Val Asp Ser Ser Val Asp Ser Tyr Cys Phe Pro Gly Gly Ala Gly Gly
260 265 270
gac gag tac cac gag tgc gtg gtg ggc ccc gtg gcg ggc ggc atc cag
864Asp Glu Tyr His Glu Cys Val Val Gly Pro Val Ala Gly Gly Ile Gln
275 280 285
tcg gag gag gac gac ggc gcg ggc agc gac gag ggc tgc agc tac tac
912Ser Glu Glu Asp Asp Gly Ala Gly Ser Asp Glu Gly Cys Ser Tyr Tyr
290 295 300
ccc gac gac gcc gcc gtc ttc ttc gcc gcc gcg caa ggg cac ggc cac
960Pro Asp Asp Ala Ala Val Phe Phe Ala Ala Ala Gln Gly His Gly His
305 310 315 320
cat cgc acg gac gac gac gat cag cag gac gac ggc cag atc agc tac
1008His Arg Thr Asp Asp Asp Asp Gln Gln Asp Asp Gly Gln Ile Ser Tyr
325 330 335
tgg atg tgg aac tag
1023Trp Met Trp Asn
340
16340PRTTriticum aestivum 16Met Glu Pro Gly Arg Leu Ile Phe Asn Thr Ser
Gly Ser Gly Asn Gly 1 5 10
15 Gln Met Leu Phe Met Asp Cys Gly Ala Gly Gly Ile Ala Gly Ala Ala
20 25 30 Gly Met
Phe His Arg Gly Val Arg Pro Val Leu Gly Gly Met Glu Glu 35
40 45 Gly Arg Gly Val Lys Arg Pro
Phe Phe Thr Ser Pro Asp Asp Met Leu 50 55
60 Glu Glu Glu Tyr Tyr Asp Glu Gln Leu Pro Glu Lys
Lys Arg Arg Leu 65 70 75
80 Thr Pro Glu Gln Val His Leu Leu Glu Arg Ser Phe Glu Glu Glu Asn
85 90 95 Lys Leu Glu
Pro Glu Arg Lys Thr Glu Leu Ala Arg Lys Leu Gly Leu 100
105 110 Gln Pro Arg Gln Val Ala Val Trp
Phe Gln Asn Arg Arg Ala Arg Trp 115 120
125 Lys Thr Lys Thr Leu Glu Arg Asp Phe Asp Arg Leu Lys
Ala Ser Phe 130 135 140
Asp Ala Leu Arg Ala Asp His Asp Ala Leu Leu Gln Asp Asn His Arg 145
150 155 160 Leu Arg Ser Gln
Val Val Thr Leu Thr Glu Lys Met Gln Asp Lys Glu 165
170 175 Ala Pro Glu Gly Ser Phe Gly Ala Ala
Ala Asp Ala Ser Glu Pro Glu 180 185
190 Gln Ala Ala Ala Glu Ala Lys Ala Ser Leu Ala Asp Ala Glu
Glu Gln 195 200 205
Ala Ala Ala Ala Glu Ala Phe Glu Val Val Gln Gln Gln Leu His Val 210
215 220 Lys Asp Glu Glu Arg
Leu Ser Pro Gly Ser Gly Gly Ser Ala Val Leu 225 230
235 240 Asp Ala Arg Asp Ala Leu Leu Gly Ser Gly
Cys Gly Leu Ala Gly Val 245 250
255 Val Asp Ser Ser Val Asp Ser Tyr Cys Phe Pro Gly Gly Ala Gly
Gly 260 265 270 Asp
Glu Tyr His Glu Cys Val Val Gly Pro Val Ala Gly Gly Ile Gln 275
280 285 Ser Glu Glu Asp Asp Gly
Ala Gly Ser Asp Glu Gly Cys Ser Tyr Tyr 290 295
300 Pro Asp Asp Ala Ala Val Phe Phe Ala Ala Ala
Gln Gly His Gly His 305 310 315
320 His Arg Thr Asp Asp Asp Asp Gln Gln Asp Asp Gly Gln Ile Ser Tyr
325 330 335 Trp Met
Trp Asn 340 17534DNABrassica napusCDS(1)..(534)zinc finger
protein containing an A20 domain and an AN1 domain (BN47310186)
17atg gac cac gac aaa aca gga tgc caa agc cca cct gaa ggt ccc aag
48Met Asp His Asp Lys Thr Gly Cys Gln Ser Pro Pro Glu Gly Pro Lys
1 5 10 15
cta tgc atc aac aac tgc ggt ttc ttc gga agc gct gcc aca atg aac
96Leu Cys Ile Asn Asn Cys Gly Phe Phe Gly Ser Ala Ala Thr Met Asn
20 25 30
atg tgt tcc aag tgt cac aag gct atc ctg ttt caa cag gaa cag ggg
144Met Cys Ser Lys Cys His Lys Ala Ile Leu Phe Gln Gln Glu Gln Gly
35 40 45
gct agg ttt gca tct gca gtg tct ggt ggt aca tca tca tcc agc aac
192Ala Arg Phe Ala Ser Ala Val Ser Gly Gly Thr Ser Ser Ser Ser Asn
50 55 60
atc tta aag gaa acc ttt gct gct acc gcg ctg gtt gat gct gaa acc
240Ile Leu Lys Glu Thr Phe Ala Ala Thr Ala Leu Val Asp Ala Glu Thr
65 70 75 80
aaa tcc gtt gag ccg gtg gct gtc tct gta cag cca tct tct gtc caa
288Lys Ser Val Glu Pro Val Ala Val Ser Val Gln Pro Ser Ser Val Gln
85 90 95
gtt gcc gca gag gta gta gct cca gaa gcc gct gca gca aaa cta aag
336Val Ala Ala Glu Val Val Ala Pro Glu Ala Ala Ala Ala Lys Leu Lys
100 105 110
gaa gga cca agc cga tgt gct act tgc aat aaa cgg gtt ggt ctg act
384Glu Gly Pro Ser Arg Cys Ala Thr Cys Asn Lys Arg Val Gly Leu Thr
115 120 125
gga ttc aaa tgt cgc tgt ggt gac ctc ttc tgc ggg acg cac cgt tat
432Gly Phe Lys Cys Arg Cys Gly Asp Leu Phe Cys Gly Thr His Arg Tyr
130 135 140
gca gac ata cac aac tgc tcc ttc aat tac cat gcc gct gcg caa gaa
480Ala Asp Ile His Asn Cys Ser Phe Asn Tyr His Ala Ala Ala Gln Glu
145 150 155 160
gct ata gct aaa gca aac ccg gtt gtg aag gca gag aag ctt gac aaa
528Ala Ile Ala Lys Ala Asn Pro Val Val Lys Ala Glu Lys Leu Asp Lys
165 170 175
atc tga
534Ile
18177PRTBrassica napus 18Met Asp His Asp Lys Thr Gly Cys Gln Ser Pro Pro
Glu Gly Pro Lys 1 5 10
15 Leu Cys Ile Asn Asn Cys Gly Phe Phe Gly Ser Ala Ala Thr Met Asn
20 25 30 Met Cys Ser
Lys Cys His Lys Ala Ile Leu Phe Gln Gln Glu Gln Gly 35
40 45 Ala Arg Phe Ala Ser Ala Val Ser
Gly Gly Thr Ser Ser Ser Ser Asn 50 55
60 Ile Leu Lys Glu Thr Phe Ala Ala Thr Ala Leu Val Asp
Ala Glu Thr 65 70 75
80 Lys Ser Val Glu Pro Val Ala Val Ser Val Gln Pro Ser Ser Val Gln
85 90 95 Val Ala Ala Glu
Val Val Ala Pro Glu Ala Ala Ala Ala Lys Leu Lys 100
105 110 Glu Gly Pro Ser Arg Cys Ala Thr Cys
Asn Lys Arg Val Gly Leu Thr 115 120
125 Gly Phe Lys Cys Arg Cys Gly Asp Leu Phe Cys Gly Thr His
Arg Tyr 130 135 140
Ala Asp Ile His Asn Cys Ser Phe Asn Tyr His Ala Ala Ala Gln Glu 145
150 155 160 Ala Ile Ala Lys Ala
Asn Pro Val Val Lys Ala Glu Lys Leu Asp Lys 165
170 175 Ile 19564DNABrassica
napusCDS(1)..(564)zinc finger protein containing an A20 domain and
an AN1 domain (BN51359456) 19atg gcg gaa gag cat cga tgc cag acg ccg gaa
ggc cac cgt ctc tgt 48Met Ala Glu Glu His Arg Cys Gln Thr Pro Glu
Gly His Arg Leu Cys 1 5 10
15 gct aac aac tgc ggc ttc ctc ggc agc tcc gcc acc
atg aat cta tgc 96Ala Asn Asn Cys Gly Phe Leu Gly Ser Ser Ala Thr
Met Asn Leu Cys 20 25
30 tcc aac tgc tac ggc gat ctc tgc ctt aag caa cag caa
gct tcc atg 144Ser Asn Cys Tyr Gly Asp Leu Cys Leu Lys Gln Gln Gln
Ala Ser Met 35 40 45
aaa tcc acc gtc gaa tcc tct ctc tcc gcc gta tct cct ccg
tcg tca 192Lys Ser Thr Val Glu Ser Ser Leu Ser Ala Val Ser Pro Pro
Ser Ser 50 55 60
gag atc ggc tct atg caa tcc acc gtt gaa tcc tct ctc tcc gac
gta 240Glu Ile Gly Ser Met Gln Ser Thr Val Glu Ser Ser Leu Ser Asp
Val 65 70 75
80 tct cct cca tca ccg gag acc att tcc atc tcc tct cca atg atc
cag 288Ser Pro Pro Ser Pro Glu Thr Ile Ser Ile Ser Ser Pro Met Ile
Gln 85 90 95
cct ctc gtt cga aac cca tca gct gaa ttg gag gta acg gcg acg aag
336Pro Leu Val Arg Asn Pro Ser Ala Glu Leu Glu Val Thr Ala Thr Lys
100 105 110
acg gtg act ccg ccg ccg gag cag cag cag aaa cgg ccg aat cgg tgc
384Thr Val Thr Pro Pro Pro Glu Gln Gln Gln Lys Arg Pro Asn Arg Cys
115 120 125
acg acg tgt agg aaa cgg gtc ggg ttg acc ggg ttc aag tgc cgg tgc
432Thr Thr Cys Arg Lys Arg Val Gly Leu Thr Gly Phe Lys Cys Arg Cys
130 135 140
ggg acg act ttt tgc ggg gct cac agg tac ccg gag gtc cat gga tgc
480Gly Thr Thr Phe Cys Gly Ala His Arg Tyr Pro Glu Val His Gly Cys
145 150 155 160
acc ttc gat ttc aaa tcg gcc ggt cgc gaa gag atc gcc aag gcg aac
528Thr Phe Asp Phe Lys Ser Ala Gly Arg Glu Glu Ile Ala Lys Ala Asn
165 170 175
cca ctc gtc aaa gcg gcg aag ctt cag aag att tga
564Pro Leu Val Lys Ala Ala Lys Leu Gln Lys Ile
180 185
20187PRTBrassica napus 20Met Ala Glu Glu His Arg Cys Gln Thr Pro Glu Gly
His Arg Leu Cys 1 5 10
15 Ala Asn Asn Cys Gly Phe Leu Gly Ser Ser Ala Thr Met Asn Leu Cys
20 25 30 Ser Asn Cys
Tyr Gly Asp Leu Cys Leu Lys Gln Gln Gln Ala Ser Met 35
40 45 Lys Ser Thr Val Glu Ser Ser Leu
Ser Ala Val Ser Pro Pro Ser Ser 50 55
60 Glu Ile Gly Ser Met Gln Ser Thr Val Glu Ser Ser Leu
Ser Asp Val 65 70 75
80 Ser Pro Pro Ser Pro Glu Thr Ile Ser Ile Ser Ser Pro Met Ile Gln
85 90 95 Pro Leu Val Arg
Asn Pro Ser Ala Glu Leu Glu Val Thr Ala Thr Lys 100
105 110 Thr Val Thr Pro Pro Pro Glu Gln Gln
Gln Lys Arg Pro Asn Arg Cys 115 120
125 Thr Thr Cys Arg Lys Arg Val Gly Leu Thr Gly Phe Lys Cys
Arg Cys 130 135 140
Gly Thr Thr Phe Cys Gly Ala His Arg Tyr Pro Glu Val His Gly Cys 145
150 155 160 Thr Phe Asp Phe Lys
Ser Ala Gly Arg Glu Glu Ile Ala Lys Ala Asn 165
170 175 Pro Leu Val Lys Ala Ala Lys Leu Gln Lys
Ile 180 185 21465DNAHordeum
vulgareCDS(1)..(465)zinc finger protein containing an A20 domain and
an AN1 domain (HV62552639) 21atg gcc cag gag agt tgt gat ctc aac aag gac
gag gcc gag atc ctg 48Met Ala Gln Glu Ser Cys Asp Leu Asn Lys Asp
Glu Ala Glu Ile Leu 1 5 10
15 aag cca tcc tcc tcc aca cct tcg cct cct tcg cca
gcc aca cca cca 96Lys Pro Ser Ser Ser Thr Pro Ser Pro Pro Ser Pro
Ala Thr Pro Pro 20 25
30 cca cca acc gct caa ata cca gaa cca caa cct cca cac
tca cca cca 144Pro Pro Thr Ala Gln Ile Pro Glu Pro Gln Pro Pro His
Ser Pro Pro 35 40 45
caa cca ccg gca gct caa ttc ttg tcc agg ccc tgc gag gtt
gtt ccc 192Gln Pro Pro Ala Ala Gln Phe Leu Ser Arg Pro Cys Glu Val
Val Pro 50 55 60
ata gag act tcc aaa aag agg aaa cat gct gat gcg gtg tca atg
gcc 240Ile Glu Thr Ser Lys Lys Arg Lys His Ala Asp Ala Val Ser Met
Ala 65 70 75
80 att gtg gtt gag cca ttg tcg tct gtg ctg ttc gtt aac cgt tgc
aac 288Ile Val Val Glu Pro Leu Ser Ser Val Leu Phe Val Asn Arg Cys
Asn 85 90 95
gtg tgc cgc aag aga gtt ggt ttg acc ggg ttc cgt tgc cgg tgt gag
336Val Cys Arg Lys Arg Val Gly Leu Thr Gly Phe Arg Cys Arg Cys Glu
100 105 110
aag ctc ttt tgt ccg cgc cac cgg cat tca gaa agc cac gac tgc tca
384Lys Leu Phe Cys Pro Arg His Arg His Ser Glu Ser His Asp Cys Ser
115 120 125
ttt gat tat aaa act gtg ggt cgg gag gag att gcc cgg gca aac cct
432Phe Asp Tyr Lys Thr Val Gly Arg Glu Glu Ile Ala Arg Ala Asn Pro
130 135 140
ctg atc agg gct gcc aag atc att agg ata tga
465Leu Ile Arg Ala Ala Lys Ile Ile Arg Ile
145 150
22154PRTHordeum vulgare 22Met Ala Gln Glu Ser Cys Asp Leu Asn Lys Asp Glu
Ala Glu Ile Leu 1 5 10
15 Lys Pro Ser Ser Ser Thr Pro Ser Pro Pro Ser Pro Ala Thr Pro Pro
20 25 30 Pro Pro Thr
Ala Gln Ile Pro Glu Pro Gln Pro Pro His Ser Pro Pro 35
40 45 Gln Pro Pro Ala Ala Gln Phe Leu
Ser Arg Pro Cys Glu Val Val Pro 50 55
60 Ile Glu Thr Ser Lys Lys Arg Lys His Ala Asp Ala Val
Ser Met Ala 65 70 75
80 Ile Val Val Glu Pro Leu Ser Ser Val Leu Phe Val Asn Arg Cys Asn
85 90 95 Val Cys Arg Lys
Arg Val Gly Leu Thr Gly Phe Arg Cys Arg Cys Glu 100
105 110 Lys Leu Phe Cys Pro Arg His Arg His
Ser Glu Ser His Asp Cys Ser 115 120
125 Phe Asp Tyr Lys Thr Val Gly Arg Glu Glu Ile Ala Arg Ala
Asn Pro 130 135 140
Leu Ile Arg Ala Ala Lys Ile Ile Arg Ile 145 150
23516DNAZea maysCDS(1)..(516)zinc finger protein containing an A20
domain and an AN1 domain (ZM61995511) 23atg gaa cac aag gag gcg ggc
tgc cag cag ccg gag ggc cca atc cta 48Met Glu His Lys Glu Ala Gly
Cys Gln Gln Pro Glu Gly Pro Ile Leu 1 5
10 15 tgc atc aat aac tgc ggc ttc ttc
ggc agt gct gcg acg atg aac atg 96Cys Ile Asn Asn Cys Gly Phe Phe
Gly Ser Ala Ala Thr Met Asn Met 20
25 30 tgc tcc aag tgc cac aag gag atg
ata acg aag cag gag cag gcc cag 144Cys Ser Lys Cys His Lys Glu Met
Ile Thr Lys Gln Glu Gln Ala Gln 35 40
45 ctg gct gcc tcc ccc atc gat agc att
gtc aat ggc ggt gac ggc ggg 192Leu Ala Ala Ser Pro Ile Asp Ser Ile
Val Asn Gly Gly Asp Gly Gly 50 55
60 aaa gga cct gta att gct gca tct gta aat
gtg gca gtt cct caa gtt 240Lys Gly Pro Val Ile Ala Ala Ser Val Asn
Val Ala Val Pro Gln Val 65 70
75 80 gag cag aag act att gtt gtg cag ccc atg
ctt gta gct gaa acc agc 288Glu Gln Lys Thr Ile Val Val Gln Pro Met
Leu Val Ala Glu Thr Ser 85 90
95 gag gct gct gct gta atc ccc aag gcc aag gaa
ggc cca gac cgg tgc 336Glu Ala Ala Ala Val Ile Pro Lys Ala Lys Glu
Gly Pro Asp Arg Cys 100 105
110 gcg gcc tgc agg aag cgt gtt ggg ctg acg gga ttt
agc tgc cga tgc 384Ala Ala Cys Arg Lys Arg Val Gly Leu Thr Gly Phe
Ser Cys Arg Cys 115 120
125 ggg aac atg tac tgt tcg gtg cac cgc tac tcc gac
aaa cat gac tgt 432Gly Asn Met Tyr Cys Ser Val His Arg Tyr Ser Asp
Lys His Asp Cys 130 135 140
cag ttc gac tat cgg act gca gca agg gac gcg att gcc
aag gcc aat 480Gln Phe Asp Tyr Arg Thr Ala Ala Arg Asp Ala Ile Ala
Lys Ala Asn 145 150 155
160 cct gtg gtg agg gcg gag aag ctc gac aag atc tga
516Pro Val Val Arg Ala Glu Lys Leu Asp Lys Ile
165 170
24171PRTZea mays 24Met Glu His Lys Glu Ala Gly Cys Gln Gln
Pro Glu Gly Pro Ile Leu 1 5 10
15 Cys Ile Asn Asn Cys Gly Phe Phe Gly Ser Ala Ala Thr Met Asn
Met 20 25 30 Cys
Ser Lys Cys His Lys Glu Met Ile Thr Lys Gln Glu Gln Ala Gln 35
40 45 Leu Ala Ala Ser Pro Ile
Asp Ser Ile Val Asn Gly Gly Asp Gly Gly 50 55
60 Lys Gly Pro Val Ile Ala Ala Ser Val Asn Val
Ala Val Pro Gln Val 65 70 75
80 Glu Gln Lys Thr Ile Val Val Gln Pro Met Leu Val Ala Glu Thr Ser
85 90 95 Glu Ala
Ala Ala Val Ile Pro Lys Ala Lys Glu Gly Pro Asp Arg Cys 100
105 110 Ala Ala Cys Arg Lys Arg Val
Gly Leu Thr Gly Phe Ser Cys Arg Cys 115 120
125 Gly Asn Met Tyr Cys Ser Val His Arg Tyr Ser Asp
Lys His Asp Cys 130 135 140
Gln Phe Asp Tyr Arg Thr Ala Ala Arg Asp Ala Ile Ala Lys Ala Asn 145
150 155 160 Pro Val Val
Arg Ala Glu Lys Leu Asp Lys Ile 165 170
25720DNALinum usitatissimumCDS(1)..(720)zinc finger protein containing
an A20 domain and an AN1 domain (LU61567101) 25atg gct cct tca cct
tgc gtc cac ggc tgc acg gcc aat tgc ccc cgc 48Met Ala Pro Ser Pro
Cys Val His Gly Cys Thr Ala Asn Cys Pro Arg 1 5
10 15 tgc cac tct tac gga cac
ccc atc ttc ggg aac tca gat ctc gcc gct 96Cys His Ser Tyr Gly His
Pro Ile Phe Gly Asn Ser Asp Leu Ala Ala 20
25 30 ggc ggc agc gat acg tcc acg
tcg gtg ttt gga aaa gta gga tcc gtc 144Gly Gly Ser Asp Thr Ser Thr
Ser Val Phe Gly Lys Val Gly Ser Val 35
40 45 gtg att cag tcg cct gcg aag
aat cac gcg ttc ggc caa gct tgt ggc 192Val Ile Gln Ser Pro Ala Lys
Asn His Ala Phe Gly Gln Ala Cys Gly 50 55
60 ccg gtt ttt ccc tcg agc tcc tcc
cct ttc cgc cgc atc aag ttc ggc 240Pro Val Phe Pro Ser Ser Ser Ser
Pro Phe Arg Arg Ile Lys Phe Gly 65 70
75 80 ccc aaa gat ggc gag ggg aaa gga ccg
ctg aag ccg atc gag aag cag 288Pro Lys Asp Gly Glu Gly Lys Gly Pro
Leu Lys Pro Ile Glu Lys Gln 85
90 95 ccg tcg aag aag cgt ccg ttc tgc ttc
tct ccc gac gag acg att gac 336Pro Ser Lys Lys Arg Pro Phe Cys Phe
Ser Pro Asp Glu Thr Ile Asp 100 105
110 gcg acg gtt cct ccg tcc acc aaa ccg ttc
ggt tcg ttc cgt tcc gtc 384Ala Thr Val Pro Pro Ser Thr Lys Pro Phe
Gly Ser Phe Arg Ser Val 115 120
125 tgt gtc acg gac gcc gac gag gcc agg ttg aag
gcg aac cgc gag ttc 432Cys Val Thr Asp Ala Asp Glu Ala Arg Leu Lys
Ala Asn Arg Glu Phe 130 135
140 ttc gct ccg gta tcc cgc aaa cgt ggc ttc gat
ccg act gac atg acc 480Phe Ala Pro Val Ser Arg Lys Arg Gly Phe Asp
Pro Thr Asp Met Thr 145 150 155
160 ttc ggt aac gcc gcc gcc gct gcg gct aat gcg agg
gag gaa gcg aag 528Phe Gly Asn Ala Ala Ala Ala Ala Ala Asn Ala Arg
Glu Glu Ala Lys 165 170
175 aag tgg tgc ggc agt tgc aag aag cgc gtg ggg ctg tta
ggg ttc aag 576Lys Trp Cys Gly Ser Cys Lys Lys Arg Val Gly Leu Leu
Gly Phe Lys 180 185
190 tgc agg tgt acg aag ttc ttc tgt ggg aag cat cgg tat
cct gag gag 624Cys Arg Cys Thr Lys Phe Phe Cys Gly Lys His Arg Tyr
Pro Glu Glu 195 200 205
cat ggt tgt acg ttc gat cat gtg gcg ttc ggg agg cgg att
atc gag 672His Gly Cys Thr Phe Asp His Val Ala Phe Gly Arg Arg Ile
Ile Glu 210 215 220
aaa cag aat cct gtt ctc gag acc gac aag ctg gtg gac aga atc
tga 720Lys Gln Asn Pro Val Leu Glu Thr Asp Lys Leu Val Asp Arg Ile
225 230 235
26239PRTLinum usitatissimum 26Met Ala Pro Ser Pro Cys Val His Gly
Cys Thr Ala Asn Cys Pro Arg 1 5 10
15 Cys His Ser Tyr Gly His Pro Ile Phe Gly Asn Ser Asp Leu
Ala Ala 20 25 30
Gly Gly Ser Asp Thr Ser Thr Ser Val Phe Gly Lys Val Gly Ser Val
35 40 45 Val Ile Gln Ser
Pro Ala Lys Asn His Ala Phe Gly Gln Ala Cys Gly 50
55 60 Pro Val Phe Pro Ser Ser Ser Ser
Pro Phe Arg Arg Ile Lys Phe Gly 65 70
75 80 Pro Lys Asp Gly Glu Gly Lys Gly Pro Leu Lys Pro
Ile Glu Lys Gln 85 90
95 Pro Ser Lys Lys Arg Pro Phe Cys Phe Ser Pro Asp Glu Thr Ile Asp
100 105 110 Ala Thr Val
Pro Pro Ser Thr Lys Pro Phe Gly Ser Phe Arg Ser Val 115
120 125 Cys Val Thr Asp Ala Asp Glu Ala
Arg Leu Lys Ala Asn Arg Glu Phe 130 135
140 Phe Ala Pro Val Ser Arg Lys Arg Gly Phe Asp Pro Thr
Asp Met Thr 145 150 155
160 Phe Gly Asn Ala Ala Ala Ala Ala Ala Asn Ala Arg Glu Glu Ala Lys
165 170 175 Lys Trp Cys Gly
Ser Cys Lys Lys Arg Val Gly Leu Leu Gly Phe Lys 180
185 190 Cys Arg Cys Thr Lys Phe Phe Cys Gly
Lys His Arg Tyr Pro Glu Glu 195 200
205 His Gly Cys Thr Phe Asp His Val Ala Phe Gly Arg Arg Ile
Ile Glu 210 215 220
Lys Gln Asn Pro Val Leu Glu Thr Asp Lys Leu Val Asp Arg Ile 225
230 235 27510DNALinum
usitatissimumCDS(1)..(510)zinc finger protein containing an A20 domain
and an AN1 domain (LU61893412) 27atg gac cat gac gag gca ggc tgc cag
gct cct tcc gat cat cct att 48Met Asp His Asp Glu Ala Gly Cys Gln
Ala Pro Ser Asp His Pro Ile 1 5
10 15 ctg tgc gtt aac aat tgc ggc ttc ttc
gga agt gct gcc acc atg aac 96Leu Cys Val Asn Asn Cys Gly Phe Phe
Gly Ser Ala Ala Thr Met Asn 20 25
30 atg tgc tca aag tgc cac aag gat acg atg
cta aac caa gag caa tcc 144Met Cys Ser Lys Cys His Lys Asp Thr Met
Leu Asn Gln Glu Gln Ser 35 40
45 aag ctt gct gct tca tcg gca gca agt atc ctc
aac gga tcg tcg atg 192Lys Leu Ala Ala Ser Ser Ala Ala Ser Ile Leu
Asn Gly Ser Ser Met 50 55
60 agc ctc gga agg gaa ctc gtt att gct gct aag
acc aat tcg gta gaa 240Ser Leu Gly Arg Glu Leu Val Ile Ala Ala Lys
Thr Asn Ser Val Glu 65 70 75
80 ccc aag acc atc tcc gtc caa cca tct tct gct tca
agt gct gaa gag 288Pro Lys Thr Ile Ser Val Gln Pro Ser Ser Ala Ser
Ser Ala Glu Glu 85 90
95 agt atc gaa atg aag ctg cca aaa gaa ggg ccc agt agg
tgc aac act 336Ser Ile Glu Met Lys Leu Pro Lys Glu Gly Pro Ser Arg
Cys Asn Thr 100 105
110 tgc aac aaa cgt gtc ggt ttg acc gga ttc aaa tgt cgg
tgc gag aac 384Cys Asn Lys Arg Val Gly Leu Thr Gly Phe Lys Cys Arg
Cys Glu Asn 115 120 125
atg ttc tgc gca aac cat cgc tac tcg gac aag cac aat tgc
ccc ttt 432Met Phe Cys Ala Asn His Arg Tyr Ser Asp Lys His Asn Cys
Pro Phe 130 135 140
gat tac cgc act gct ggc cgt gaa gct atc tca aag gcc aat cct
ttg 480Asp Tyr Arg Thr Ala Gly Arg Glu Ala Ile Ser Lys Ala Asn Pro
Leu 145 150 155
160 gtg aag gcg gag aag ctc gac aaa atc tga
510Val Lys Ala Glu Lys Leu Asp Lys Ile
165
28169PRTLinum usitatissimum 28Met Asp His Asp Glu Ala Gly Cys Gln
Ala Pro Ser Asp His Pro Ile 1 5 10
15 Leu Cys Val Asn Asn Cys Gly Phe Phe Gly Ser Ala Ala Thr
Met Asn 20 25 30
Met Cys Ser Lys Cys His Lys Asp Thr Met Leu Asn Gln Glu Gln Ser
35 40 45 Lys Leu Ala Ala
Ser Ser Ala Ala Ser Ile Leu Asn Gly Ser Ser Met 50
55 60 Ser Leu Gly Arg Glu Leu Val Ile
Ala Ala Lys Thr Asn Ser Val Glu 65 70
75 80 Pro Lys Thr Ile Ser Val Gln Pro Ser Ser Ala Ser
Ser Ala Glu Glu 85 90
95 Ser Ile Glu Met Lys Leu Pro Lys Glu Gly Pro Ser Arg Cys Asn Thr
100 105 110 Cys Asn Lys
Arg Val Gly Leu Thr Gly Phe Lys Cys Arg Cys Glu Asn 115
120 125 Met Phe Cys Ala Asn His Arg Tyr
Ser Asp Lys His Asn Cys Pro Phe 130 135
140 Asp Tyr Arg Thr Ala Gly Arg Glu Ala Ile Ser Lys Ala
Asn Pro Leu 145 150 155
160 Val Lys Ala Glu Lys Leu Asp Lys Ile 165
29495DNAOryza sativaCDS(1)..(495)zinc finger protein containing an A20
domain and an AN1 domain (OS39781852) 29atg gcg cag cgc gac aag aag
gat cag gag ccg acg gag ctc agg gcg 48Met Ala Gln Arg Asp Lys Lys
Asp Gln Glu Pro Thr Glu Leu Arg Ala 1 5
10 15 ccg gag atc acg ctg tgc gcc aac
agc tgc gga ttc ccg ggc aac ccg 96Pro Glu Ile Thr Leu Cys Ala Asn
Ser Cys Gly Phe Pro Gly Asn Pro 20
25 30 gcc acg cag aac ctc tgc cag aac
tgc ttc ttg gcg gcc acg gcg tcc 144Ala Thr Gln Asn Leu Cys Gln Asn
Cys Phe Leu Ala Ala Thr Ala Ser 35 40
45 acc tcg tcg ccg tct tct ttg tcg tca
ccg gtg ctc gac aag cag ccg 192Thr Ser Ser Pro Ser Ser Leu Ser Ser
Pro Val Leu Asp Lys Gln Pro 50 55
60 ccg agg ccg gcg gcg ccg ctg gtt gag cct
cag gct cct ctc cca ccg 240Pro Arg Pro Ala Ala Pro Leu Val Glu Pro
Gln Ala Pro Leu Pro Pro 65 70
75 80 cct gtg gag gag atg gcc tcc gcg ctc gcg
acg gcg ccg gcg ccg gtc 288Pro Val Glu Glu Met Ala Ser Ala Leu Ala
Thr Ala Pro Ala Pro Val 85 90
95 gcc aag acg tcg gcg gtg aac cgg tgc tcc agg
tgc cgg aag cgt gtc 336Ala Lys Thr Ser Ala Val Asn Arg Cys Ser Arg
Cys Arg Lys Arg Val 100 105
110 ggc ctc acc ggg ttc cgg tgc cgg tgc ggc cac ctg
ttc tgc ggc gag 384Gly Leu Thr Gly Phe Arg Cys Arg Cys Gly His Leu
Phe Cys Gly Glu 115 120
125 cac cgg tac tcc gac cgc cac ggc tgc agc tac gac
tac aag tcg gcg 432His Arg Tyr Ser Asp Arg His Gly Cys Ser Tyr Asp
Tyr Lys Ser Ala 130 135 140
gcg agg gac gcc atc gcc agg gac aac ccg gtg gtg cgc
gcg gcc aag 480Ala Arg Asp Ala Ile Ala Arg Asp Asn Pro Val Val Arg
Ala Ala Lys 145 150 155
160 atc gtt agg ttc tga
495Ile Val Arg Phe
30164PRTOryza sativa 30Met Ala Gln Arg Asp Lys Lys Asp Gln
Glu Pro Thr Glu Leu Arg Ala 1 5 10
15 Pro Glu Ile Thr Leu Cys Ala Asn Ser Cys Gly Phe Pro Gly
Asn Pro 20 25 30
Ala Thr Gln Asn Leu Cys Gln Asn Cys Phe Leu Ala Ala Thr Ala Ser
35 40 45 Thr Ser Ser Pro
Ser Ser Leu Ser Ser Pro Val Leu Asp Lys Gln Pro 50
55 60 Pro Arg Pro Ala Ala Pro Leu Val
Glu Pro Gln Ala Pro Leu Pro Pro 65 70
75 80 Pro Val Glu Glu Met Ala Ser Ala Leu Ala Thr Ala
Pro Ala Pro Val 85 90
95 Ala Lys Thr Ser Ala Val Asn Arg Cys Ser Arg Cys Arg Lys Arg Val
100 105 110 Gly Leu Thr
Gly Phe Arg Cys Arg Cys Gly His Leu Phe Cys Gly Glu 115
120 125 His Arg Tyr Ser Asp Arg His Gly
Cys Ser Tyr Asp Tyr Lys Ser Ala 130 135
140 Ala Arg Asp Ala Ile Ala Arg Asp Asn Pro Val Val Arg
Ala Ala Lys 145 150 155
160 Ile Val Arg Phe 31495DNAOryza sativaCDS(1)..(495)zinc finger protein
containing an A20 domain and an AN1 domain (OS34701560) 31atg gcc
gaa gaa cac cga tgc caa gct ccc gaa ggt cac aga ctc tgc 48Met Ala
Glu Glu His Arg Cys Gln Ala Pro Glu Gly His Arg Leu Cys 1
5 10 15 tcc aac aac
tgc ggt ttc ttt ggt agc ccc gcc acc atg aat ctc tgt 96Ser Asn Asn
Cys Gly Phe Phe Gly Ser Pro Ala Thr Met Asn Leu Cys
20 25 30 tcc aaa tgc
tac aga gac atc cgt ttg aag gaa gaa gaa caa gcc aaa 144Ser Lys Cys
Tyr Arg Asp Ile Arg Leu Lys Glu Glu Glu Gln Ala Lys 35
40 45 acc aaa tcc aca
atc gaa acc gct ctt tca gga tct tcc tcc gcc acc 192Thr Lys Ser Thr
Ile Glu Thr Ala Leu Ser Gly Ser Ser Ser Ala Thr 50
55 60 gtc acc gca acc gcc
gtc gtt gcc tcc tcc gtg gaa tcc cct tcg gcg 240Val Thr Ala Thr Ala
Val Val Ala Ser Ser Val Glu Ser Pro Ser Ala 65
70 75 80 ccg gtt gaa tcc ctc
cct caa cca ccg gtg ctg att tcg ccg gat ata 288Pro Val Glu Ser Leu
Pro Gln Pro Pro Val Leu Ile Ser Pro Asp Ile 85
90 95 gcc gca ccg gtt cag gcg
aac cgg tgc ggc gcg tgt agg aag cgc gtg 336Ala Ala Pro Val Gln Ala
Asn Arg Cys Gly Ala Cys Arg Lys Arg Val 100
105 110 ggg ttg aca ggg ttc aag tgc
agg tgc gga aca acg ttt tgt ggg agc 384Gly Leu Thr Gly Phe Lys Cys
Arg Cys Gly Thr Thr Phe Cys Gly Ser 115
120 125 cac agg tac ccc gag aaa cac
gcg tgt ggc ttc gat ttc aag gcg gtg 432His Arg Tyr Pro Glu Lys His
Ala Cys Gly Phe Asp Phe Lys Ala Val 130 135
140 ggg aga gag gag ata gca cgg gcg
aat ccc gtg atc aaa ggc gag aag 480Gly Arg Glu Glu Ile Ala Arg Ala
Asn Pro Val Ile Lys Gly Glu Lys 145 150
155 160 cta cgg agg att taa
495Leu Arg Arg Ile
32164PRTOryza sativa 32Met Ala Glu Glu
His Arg Cys Gln Ala Pro Glu Gly His Arg Leu Cys 1 5
10 15 Ser Asn Asn Cys Gly Phe Phe Gly Ser
Pro Ala Thr Met Asn Leu Cys 20 25
30 Ser Lys Cys Tyr Arg Asp Ile Arg Leu Lys Glu Glu Glu Gln
Ala Lys 35 40 45
Thr Lys Ser Thr Ile Glu Thr Ala Leu Ser Gly Ser Ser Ser Ala Thr 50
55 60 Val Thr Ala Thr Ala
Val Val Ala Ser Ser Val Glu Ser Pro Ser Ala 65 70
75 80 Pro Val Glu Ser Leu Pro Gln Pro Pro Val
Leu Ile Ser Pro Asp Ile 85 90
95 Ala Ala Pro Val Gln Ala Asn Arg Cys Gly Ala Cys Arg Lys Arg
Val 100 105 110 Gly
Leu Thr Gly Phe Lys Cys Arg Cys Gly Thr Thr Phe Cys Gly Ser 115
120 125 His Arg Tyr Pro Glu Lys
His Ala Cys Gly Phe Asp Phe Lys Ala Val 130 135
140 Gly Arg Glu Glu Ile Ala Arg Ala Asn Pro Val
Ile Lys Gly Glu Lys 145 150 155
160 Leu Arg Arg Ile 33513DNAOryza sativaCDS(1)..(513)zinc finger
protein containing an A20 domain and an AN1 domain (OS36821256)
33atg gcg cag agg gag aag aag gtg gag gag ccg acg gag ctg agg gcg
48Met Ala Gln Arg Glu Lys Lys Val Glu Glu Pro Thr Glu Leu Arg Ala
1 5 10 15
ccg gag atg acg ctc tgc gcc aac agc tgc ggg ttc ccg ggc aac ccg
96Pro Glu Met Thr Leu Cys Ala Asn Ser Cys Gly Phe Pro Gly Asn Pro
20 25 30
gcg acc aac aac ctc tgc cag aac tgc ttc ttg gct gcc tcg gcg tct
144Ala Thr Asn Asn Leu Cys Gln Asn Cys Phe Leu Ala Ala Ser Ala Ser
35 40 45
tct tct tct tct tcc gcc gct gcc tcg ccg tcg acg acg tcg ttg ccg
192Ser Ser Ser Ser Ser Ala Ala Ala Ser Pro Ser Thr Thr Ser Leu Pro
50 55 60
gtg ttt ccg gtg gtg gag aag ccg agg cag gcc gta cag tcg tcg gcg
240Val Phe Pro Val Val Glu Lys Pro Arg Gln Ala Val Gln Ser Ser Ala
65 70 75 80
gcg gcg gcg gtg gcg ctg gtg gtt gag cgg ccg acg gcg ggg ccg gtg
288Ala Ala Ala Val Ala Leu Val Val Glu Arg Pro Thr Ala Gly Pro Val
85 90 95
gag tcg tcg tcg aag gcg tcg agg tcg tcg tcg gtc aac cga tgc cac
336Glu Ser Ser Ser Lys Ala Ser Arg Ser Ser Ser Val Asn Arg Cys His
100 105 110
agc tgc cgg agg cgg gtg ggc ctg acc ggg ttc cgg tgc cgc tgc ggc
384Ser Cys Arg Arg Arg Val Gly Leu Thr Gly Phe Arg Cys Arg Cys Gly
115 120 125
gag ctc tac tgc ggc gcg cac cgg tac tcc gac cgc cac gac tgc agc
432Glu Leu Tyr Cys Gly Ala His Arg Tyr Ser Asp Arg His Asp Cys Ser
130 135 140
ttc gac tac aag tcg gcg gcg agg gac gcc atc gcc agg gag aac ccc
480Phe Asp Tyr Lys Ser Ala Ala Arg Asp Ala Ile Ala Arg Glu Asn Pro
145 150 155 160
gtc gtc cgc gcc gcc aag atc gtt agg ttc taa
513Val Val Arg Ala Ala Lys Ile Val Arg Phe
165 170
34170PRTOryza sativa 34Met Ala Gln Arg Glu Lys Lys Val Glu Glu Pro Thr
Glu Leu Arg Ala 1 5 10
15 Pro Glu Met Thr Leu Cys Ala Asn Ser Cys Gly Phe Pro Gly Asn Pro
20 25 30 Ala Thr Asn
Asn Leu Cys Gln Asn Cys Phe Leu Ala Ala Ser Ala Ser 35
40 45 Ser Ser Ser Ser Ser Ala Ala Ala
Ser Pro Ser Thr Thr Ser Leu Pro 50 55
60 Val Phe Pro Val Val Glu Lys Pro Arg Gln Ala Val Gln
Ser Ser Ala 65 70 75
80 Ala Ala Ala Val Ala Leu Val Val Glu Arg Pro Thr Ala Gly Pro Val
85 90 95 Glu Ser Ser Ser
Lys Ala Ser Arg Ser Ser Ser Val Asn Arg Cys His 100
105 110 Ser Cys Arg Arg Arg Val Gly Leu Thr
Gly Phe Arg Cys Arg Cys Gly 115 120
125 Glu Leu Tyr Cys Gly Ala His Arg Tyr Ser Asp Arg His Asp
Cys Ser 130 135 140
Phe Asp Tyr Lys Ser Ala Ala Arg Asp Ala Ile Ala Arg Glu Asn Pro 145
150 155 160 Val Val Arg Ala Ala
Lys Ile Val Arg Phe 165 170
35486DNAGlycine maxCDS(1)..(486)zinc finger protein containing an A20
domain and an AN1 domain (GM51659494) 35atg gct cag aaa acc gag aaa
gaa gaa acc gac ttc aaa gtt ccg gaa 48Met Ala Gln Lys Thr Glu Lys
Glu Glu Thr Asp Phe Lys Val Pro Glu 1 5
10 15 acg att acg ctt tgc gtc aac aac
tgc ggc gtc acc gga aac cct gcc 96Thr Ile Thr Leu Cys Val Asn Asn
Cys Gly Val Thr Gly Asn Pro Ala 20
25 30 acg aat aac atg tgc cag aag tgc
ttc act gcc tct acc gcc acc act 144Thr Asn Asn Met Cys Gln Lys Cys
Phe Thr Ala Ser Thr Ala Thr Thr 35 40
45 tcc ggc gcc gga ggt gcc gga ata gct
tct ccg gcg acc aga tcc ggc 192Ser Gly Ala Gly Gly Ala Gly Ile Ala
Ser Pro Ala Thr Arg Ser Gly 50 55
60 gtc tcc gcg cgt cct cag aag aga tct ttt
cct gaa gag ccc tcg ccg 240Val Ser Ala Arg Pro Gln Lys Arg Ser Phe
Pro Glu Glu Pro Ser Pro 65 70
75 80 gtg gcg gat cct cct tct tcg gac cag acg
acg ccg tcg gag gcg aag 288Val Ala Asp Pro Pro Ser Ser Asp Gln Thr
Thr Pro Ser Glu Ala Lys 85 90
95 cgc gtg gtc aac cgc tgc tcc gga tgc cgg cgg
aag gtc gga ctc acc 336Arg Val Val Asn Arg Cys Ser Gly Cys Arg Arg
Lys Val Gly Leu Thr 100 105
110 gga ttc cgg tgc cgg tgc ggc gag ctc ttc tgc gcc
gag cac cgg tac 384Gly Phe Arg Cys Arg Cys Gly Glu Leu Phe Cys Ala
Glu His Arg Tyr 115 120
125 tcc gac cgc cac gac tgc agc tat gac tac aaa gcc
gcc gga aga gaa 432Ser Asp Arg His Asp Cys Ser Tyr Asp Tyr Lys Ala
Ala Gly Arg Glu 130 135 140
gcc atc gcg agg gag aat ccg gtg atc aga gct gcg aag
atc gtc aaa 480Ala Ile Ala Arg Glu Asn Pro Val Ile Arg Ala Ala Lys
Ile Val Lys 145 150 155
160 gtc tga
486Val
36161PRTGlycine max 36Met Ala Gln Lys Thr Glu Lys Glu Glu
Thr Asp Phe Lys Val Pro Glu 1 5 10
15 Thr Ile Thr Leu Cys Val Asn Asn Cys Gly Val Thr Gly Asn
Pro Ala 20 25 30
Thr Asn Asn Met Cys Gln Lys Cys Phe Thr Ala Ser Thr Ala Thr Thr
35 40 45 Ser Gly Ala Gly
Gly Ala Gly Ile Ala Ser Pro Ala Thr Arg Ser Gly 50
55 60 Val Ser Ala Arg Pro Gln Lys Arg
Ser Phe Pro Glu Glu Pro Ser Pro 65 70
75 80 Val Ala Asp Pro Pro Ser Ser Asp Gln Thr Thr Pro
Ser Glu Ala Lys 85 90
95 Arg Val Val Asn Arg Cys Ser Gly Cys Arg Arg Lys Val Gly Leu Thr
100 105 110 Gly Phe Arg
Cys Arg Cys Gly Glu Leu Phe Cys Ala Glu His Arg Tyr 115
120 125 Ser Asp Arg His Asp Cys Ser Tyr
Asp Tyr Lys Ala Ala Gly Arg Glu 130 135
140 Ala Ile Ala Arg Glu Asn Pro Val Ile Arg Ala Ala Lys
Ile Val Lys 145 150 155
160 Val 37519DNAGlycine maxCDS(1)..(519)zinc finger protein containing an
A20 domain and an AN1 domain (GM49780101) 37atg gag cct cat gat gag
act gga tgc cag gct cct gaa cgc ccc att 48Met Glu Pro His Asp Glu
Thr Gly Cys Gln Ala Pro Glu Arg Pro Ile 1 5
10 15 ctt tgc att aat aat tgt ggc
ttc ttt gga aga gca gct acc atg aac 96Leu Cys Ile Asn Asn Cys Gly
Phe Phe Gly Arg Ala Ala Thr Met Asn 20
25 30 atg tgt tcc aag tgt tac aag gac
atg ctg ttg aag cag gag cag gac 144Met Cys Ser Lys Cys Tyr Lys Asp
Met Leu Leu Lys Gln Glu Gln Asp 35 40
45 aaa ttt gca gca tca tcc gtt gaa aac
att gtg aat ggc agt tcc aat 192Lys Phe Ala Ala Ser Ser Val Glu Asn
Ile Val Asn Gly Ser Ser Asn 50 55
60 ggc aat gga aag cag gct gtg gct act ggt
gct gtt gct gta caa gtt 240Gly Asn Gly Lys Gln Ala Val Ala Thr Gly
Ala Val Ala Val Gln Val 65 70
75 80 gaa gct gtg gag gtc aag att gtc tgt gct
cag agt tct gtg gat tcg 288Glu Ala Val Glu Val Lys Ile Val Cys Ala
Gln Ser Ser Val Asp Ser 85 90
95 tcc tcc ggt gat agt ttg gag atg aaa gcc aag
act ggt ccc agt aga 336Ser Ser Gly Asp Ser Leu Glu Met Lys Ala Lys
Thr Gly Pro Ser Arg 100 105
110 tgt gct aca tgc cgg aaa cgt gtt ggt tta act ggt
ttc agc tgc aaa 384Cys Ala Thr Cys Arg Lys Arg Val Gly Leu Thr Gly
Phe Ser Cys Lys 115 120
125 tgt ggc aac ctc ttc tgt gca atg cat cgc tat tct
gat aaa cat gat 432Cys Gly Asn Leu Phe Cys Ala Met His Arg Tyr Ser
Asp Lys His Asp 130 135 140
tgc cct ttt gat tat agg act gtt ggt cag gat gcc ata
gct aaa gcc 480Cys Pro Phe Asp Tyr Arg Thr Val Gly Gln Asp Ala Ile
Ala Lys Ala 145 150 155
160 aac ccc ata att aag gca gat aag ctc gac aaa atc tag
519Asn Pro Ile Ile Lys Ala Asp Lys Leu Asp Lys Ile
165 170
38172PRTGlycine max 38Met Glu Pro His Asp Glu Thr Gly Cys Gln
Ala Pro Glu Arg Pro Ile 1 5 10
15 Leu Cys Ile Asn Asn Cys Gly Phe Phe Gly Arg Ala Ala Thr Met
Asn 20 25 30 Met
Cys Ser Lys Cys Tyr Lys Asp Met Leu Leu Lys Gln Glu Gln Asp 35
40 45 Lys Phe Ala Ala Ser Ser
Val Glu Asn Ile Val Asn Gly Ser Ser Asn 50 55
60 Gly Asn Gly Lys Gln Ala Val Ala Thr Gly Ala
Val Ala Val Gln Val 65 70 75
80 Glu Ala Val Glu Val Lys Ile Val Cys Ala Gln Ser Ser Val Asp Ser
85 90 95 Ser Ser
Gly Asp Ser Leu Glu Met Lys Ala Lys Thr Gly Pro Ser Arg 100
105 110 Cys Ala Thr Cys Arg Lys Arg
Val Gly Leu Thr Gly Phe Ser Cys Lys 115 120
125 Cys Gly Asn Leu Phe Cys Ala Met His Arg Tyr Ser
Asp Lys His Asp 130 135 140
Cys Pro Phe Asp Tyr Arg Thr Val Gly Gln Asp Ala Ile Ala Lys Ala 145
150 155 160 Asn Pro Ile
Ile Lys Ala Asp Lys Leu Asp Lys Ile 165
170 39525DNAGlycine maxCDS(1)..(525)zinc finger protein
containing an A20 domain and an AN1 domain (GM59637305) 39atg gac
cat gac aag act ggg tgc caa gct cct cct gaa ggt cct ata 48Met Asp
His Asp Lys Thr Gly Cys Gln Ala Pro Pro Glu Gly Pro Ile 1
5 10 15 ttg tgc atc
aac aac tgt ggg ttt ttt gga agt gca gct acc atg aac 96Leu Cys Ile
Asn Asn Cys Gly Phe Phe Gly Ser Ala Ala Thr Met Asn
20 25 30 atg tgt tct
aaa tgc cac aaa gac ata ttg ctg aaa cag gag cag gcc 144Met Cys Ser
Lys Cys His Lys Asp Ile Leu Leu Lys Gln Glu Gln Ala 35
40 45 aag ctt gca gca
tca tcc att ggg aat att atg aat ggg tca tca agc 192Lys Leu Ala Ala
Ser Ser Ile Gly Asn Ile Met Asn Gly Ser Ser Ser 50
55 60 agc act gaa aag gaa
cct gtt gtt gct gct gct gct aat att gat atc 240Ser Thr Glu Lys Glu
Pro Val Val Ala Ala Ala Ala Asn Ile Asp Ile 65
70 75 80 cca gtt att cca gta
gag cct aaa act gtc tct gtg caa cct tta ttt 288Pro Val Ile Pro Val
Glu Pro Lys Thr Val Ser Val Gln Pro Leu Phe 85
90 95 ggt tca ggt cca gag ggg
agt gtt gag gca aag ccg aag gat gga cca 336Gly Ser Gly Pro Glu Gly
Ser Val Glu Ala Lys Pro Lys Asp Gly Pro 100
105 110 aaa cgt tgc agc agc tgc aac
aag cga gtt ggt ttg aca ggg ttt aat 384Lys Arg Cys Ser Ser Cys Asn
Lys Arg Val Gly Leu Thr Gly Phe Asn 115
120 125 tgt cga tgt ggt gac ctt ttt
ttg tgc tgt aca tcg cta ctc gac aag 432Cys Arg Cys Gly Asp Leu Phe
Leu Cys Cys Thr Ser Leu Leu Asp Lys 130 135
140 cat aat tgc cca ttt gat tac cgc
act gcc gct caa gat gct ata gct 480His Asn Cys Pro Phe Asp Tyr Arg
Thr Ala Ala Gln Asp Ala Ile Ala 145 150
155 160 aaa gca aac cca gtt gtc aag gct gaa
aag ctt gat aag atc taa 525Lys Ala Asn Pro Val Val Lys Ala Glu
Lys Leu Asp Lys Ile 165
170 40174PRTGlycine max 40Met Asp His Asp
Lys Thr Gly Cys Gln Ala Pro Pro Glu Gly Pro Ile 1 5
10 15 Leu Cys Ile Asn Asn Cys Gly Phe Phe
Gly Ser Ala Ala Thr Met Asn 20 25
30 Met Cys Ser Lys Cys His Lys Asp Ile Leu Leu Lys Gln Glu
Gln Ala 35 40 45
Lys Leu Ala Ala Ser Ser Ile Gly Asn Ile Met Asn Gly Ser Ser Ser 50
55 60 Ser Thr Glu Lys Glu
Pro Val Val Ala Ala Ala Ala Asn Ile Asp Ile 65 70
75 80 Pro Val Ile Pro Val Glu Pro Lys Thr Val
Ser Val Gln Pro Leu Phe 85 90
95 Gly Ser Gly Pro Glu Gly Ser Val Glu Ala Lys Pro Lys Asp Gly
Pro 100 105 110 Lys
Arg Cys Ser Ser Cys Asn Lys Arg Val Gly Leu Thr Gly Phe Asn 115
120 125 Cys Arg Cys Gly Asp Leu
Phe Leu Cys Cys Thr Ser Leu Leu Asp Lys 130 135
140 His Asn Cys Pro Phe Asp Tyr Arg Thr Ala Ala
Gln Asp Ala Ile Ala 145 150 155
160 Lys Ala Asn Pro Val Val Lys Ala Glu Lys Leu Asp Lys Ile
165 170 41498DNATriticum
aestivumCDS(1)..(498)zinc finger protein containing an A20 domain
and an AN1 domain (TA55974113) 41atg gcg cag cgg gat cac aag cag gag gag
ccc acg gag ctg cgg gcg 48Met Ala Gln Arg Asp His Lys Gln Glu Glu
Pro Thr Glu Leu Arg Ala 1 5 10
15 ccg gag atc acg ctc tgc gcc aac agc tgc ggc
ttc ccg ggc aac ccg 96Pro Glu Ile Thr Leu Cys Ala Asn Ser Cys Gly
Phe Pro Gly Asn Pro 20 25
30 gcc acg cag aac ctc tgc cag aac tgc ttc ttg gcc
ggc ccg gcg tcc 144Ala Thr Gln Asn Leu Cys Gln Asn Cys Phe Leu Ala
Gly Pro Ala Ser 35 40
45 acg tcg ccg tct tcc tcc tcc tcc tcc tcc tct tct
ctg ccg ggc gtg 192Thr Ser Pro Ser Ser Ser Ser Ser Ser Ser Ser Ser
Leu Pro Gly Val 50 55 60
tcc gcg ccg acc ccc gtc atc gac agg ccg agg ccg gcg
ccg ttg gag 240Ser Ala Pro Thr Pro Val Ile Asp Arg Pro Arg Pro Ala
Pro Leu Glu 65 70 75
80 gcg gag ctg gca cgc ccc gcc gtc gac ctt gct ccg gcg acg
gag gcg 288Ala Glu Leu Ala Arg Pro Ala Val Asp Leu Ala Pro Ala Thr
Glu Ala 85 90
95 aag ccg gcg agg acg tcg gtg aac cgg tgc tcc agc tgc cgg
aag cgc 336Lys Pro Ala Arg Thr Ser Val Asn Arg Cys Ser Ser Cys Arg
Lys Arg 100 105 110
gtg ggg ctg acg ggg ttc cgg tgc cgg tgc ggc gac atg ttc tgc
ggc 384Val Gly Leu Thr Gly Phe Arg Cys Arg Cys Gly Asp Met Phe Cys
Gly 115 120 125
gag cac cgg tac tcg gac cgg cac ggg tgc agc tac gac tac aag gcc
432Glu His Arg Tyr Ser Asp Arg His Gly Cys Ser Tyr Asp Tyr Lys Ala
130 135 140
gcc gcc agg gac gcc atc gcc agg gac aac ccc gtc gtg cgc gcc gcc
480Ala Ala Arg Asp Ala Ile Ala Arg Asp Asn Pro Val Val Arg Ala Ala
145 150 155 160
aag atc gtc agg ttc tga
498Lys Ile Val Arg Phe
165
42165PRTTriticum aestivum 42Met Ala Gln Arg Asp His Lys Gln Glu Glu Pro
Thr Glu Leu Arg Ala 1 5 10
15 Pro Glu Ile Thr Leu Cys Ala Asn Ser Cys Gly Phe Pro Gly Asn Pro
20 25 30 Ala Thr
Gln Asn Leu Cys Gln Asn Cys Phe Leu Ala Gly Pro Ala Ser 35
40 45 Thr Ser Pro Ser Ser Ser Ser
Ser Ser Ser Ser Ser Leu Pro Gly Val 50 55
60 Ser Ala Pro Thr Pro Val Ile Asp Arg Pro Arg Pro
Ala Pro Leu Glu 65 70 75
80 Ala Glu Leu Ala Arg Pro Ala Val Asp Leu Ala Pro Ala Thr Glu Ala
85 90 95 Lys Pro Ala
Arg Thr Ser Val Asn Arg Cys Ser Ser Cys Arg Lys Arg 100
105 110 Val Gly Leu Thr Gly Phe Arg Cys
Arg Cys Gly Asp Met Phe Cys Gly 115 120
125 Glu His Arg Tyr Ser Asp Arg His Gly Cys Ser Tyr Asp
Tyr Lys Ala 130 135 140
Ala Ala Arg Asp Ala Ile Ala Arg Asp Asn Pro Val Val Arg Ala Ala 145
150 155 160 Lys Ile Val Arg
Phe 165 43207PRTPhyscomitrella
patensMISC_FEATURE(1)..(207)methionine sulfoxide reductase family protein
(EST65) 43Met Val Ala Glu Ser Val Leu Val Cys Arg Ser Ser Val Val
Gly Ala 1 5 10 15
Gly Leu Gln Ser Phe Val Gly Glu Gly Ala Lys Arg Glu Ser Ala Gly
20 25 30 Pro Gly Arg Ser Val
Phe Leu Gly Ala Gln Val Gln Lys Met Gly Ala 35
40 45 Gly Met Ser Ala Arg Ser Asp Val Arg
Pro Ala Ala Val Pro Lys Ala 50 55
60 Ser Gly Asp Val Ser Glu Gln Thr Asp Tyr Lys Thr Phe
Ser Asp Glu 65 70 75
80 Glu Trp Lys Lys Arg Leu Ser Gln Gln Gln Phe Tyr Val Ala Arg Lys
85 90 95 Lys Gly Thr Glu
Arg Pro Phe Thr Gly Glu Tyr Trp Asn Thr Lys Thr 100
105 110 Ala Gly Thr Tyr Leu Cys Val Cys Cys
Lys Thr Pro Leu Phe Ser Ser 115 120
125 Lys Thr Lys Phe Asp Ser Gly Thr Gly Trp Pro Ser Tyr Tyr
Asp Thr 130 135 140
Ile Gly Asp Asn Val Lys Ser His Met Asp Trp Ser Ile Pro Phe Met 145
150 155 160 Pro Arg Thr Glu Val
Val Cys Ala Val Cys Asp Ala His Leu Gly His 165
170 175 Val Phe Asp Asp Gly Pro Arg Pro Thr Gly
Lys Arg Tyr Cys Ile Asn 180 185
190 Ser Ala Ala Ile Asp Leu Lys Ala Glu Lys Gln Glu Glu Arg Asn
195 200 205
44339PRTPhyscomitrella patensMISC_FEATURE(1)..(339)homeodomain leucine
zipper protein (EST12) 44Met Val Val Pro Ser Leu Pro Ala Phe Gly Gly Gln
Asn Ala Met Leu 1 5 10
15 Arg Arg Asn Ile Asp Asn Asn Thr Asp Thr Leu Ile Ser Leu Leu Gln
20 25 30 Gly Ser Cys
Ser Pro Arg Val Ser Met Gln Gln Val Pro Arg Ser Ser 35
40 45 Glu Ser Leu Glu Asn Met Met Gly
Ala Cys Gly Gln Lys Leu Pro Tyr 50 55
60 Phe Ser Ser Phe Asp Gly Pro Ser Val Glu Glu Gln Glu
Asp Val Asp 65 70 75
80 Glu Gly Ile Asp Glu Phe Ala His His Val Glu Lys Lys Arg Arg Leu
85 90 95 Ser Leu Glu Gln
Val Arg Ser Leu Glu Arg Asn Phe Glu Val Glu Asn 100
105 110 Lys Leu Glu Pro Glu Arg Lys Met Gln
Leu Ala Lys Glu Leu Gly Leu 115 120
125 Arg Pro Arg Gln Val Ala Val Trp Phe Gln Asn Arg Arg Ala
Arg Trp 130 135 140
Lys Thr Lys Gln Leu Glu His Asp Tyr Glu Thr Leu Lys Lys Ala Tyr 145
150 155 160 Asp Arg Leu Lys Ala
Asp Phe Glu Ala Val Thr Leu Asp Thr Asn Ala 165
170 175 Leu Lys Ala Glu Val Ser Arg Leu Lys Gly
Ile Ser Asn Asp Asp Val 180 185
190 Lys Pro Ala Glu Phe Val Gln Gly Lys Cys Asp Thr Thr Ser His
Pro 195 200 205 Ala
Ser Pro Ala Gln Ser Glu Arg Ser Asp Ile Val Ser Ser Arg Asn 210
215 220 Arg Thr Thr Pro Thr Ile
His Val Asp Pro Val Ala Pro Glu Glu Ala 225 230
235 240 Gly Ala His Leu Thr Met Ser Ser Asp Ser Asn
Ser Ser Glu Val Met 245 250
255 Asp Ala Asp Ser Pro Arg Thr Ser His Thr Ser Ala Ser Arg Ser Thr
260 265 270 Leu Ser
Thr Ser Val Val Gln Pro Asp Glu Gly Leu Gly Val Ala Gln 275
280 285 Tyr Pro His Phe Ser Pro Glu
Asn Phe Val Gly Pro Asn Met Pro Glu 290 295
300 Ile Cys Ala Asp Gln Ser Leu Ala Ser Gln Val Lys
Leu Glu Glu Ile 305 310 315
320 His Ser Phe Asn Pro Asp Gln Thr Phe Leu Leu Leu Pro Asn Trp Trp
325 330 335 Asp Trp Ala
45188PRTPhyscomitrella patensMISC_FEATURE(1)..(188)zinc finger protein
containing an A20 domain and an AN1 domain (EST307) 45Met Ala Thr
Glu Arg Val Ser Gln Glu Thr Thr Ser Gln Ala Pro Glu 1 5
10 15 Gly Pro Val Met Cys Lys Asn Leu
Cys Gly Phe Phe Gly Ser Gln Ala 20 25
30 Thr Met Gly Leu Cys Ser Lys Cys Tyr Arg Glu Thr Val
Met Gln Ala 35 40 45
Lys Met Thr Ala Leu Ala Glu Gln Ala Thr Gln Ala Ala Gln Ala Thr 50
55 60 Ser Ala Thr Ala
Ala Ala Val Gln Pro Pro Ala Pro Val His Glu Thr 65 70
75 80 Lys Leu Thr Cys Glu Val Glu Arg Thr
Met Ile Val Pro His Gln Ser 85 90
95 Ser Ser Tyr Gln Gln Asp Leu Val Thr Pro Ala Ala Ala Ala
Pro Gln 100 105 110
Ala Val Lys Ser Ser Ile Ala Ala Pro Ser Arg Pro Glu Pro Asn Arg
115 120 125 Cys Gly Ser Cys
Arg Lys Arg Val Gly Leu Thr Gly Phe Lys Cys Arg 130
135 140 Cys Gly Asn Leu Tyr Cys Ala Leu
His Arg Tyr Ser Asp Lys His Thr 145 150
155 160 Cys Thr Tyr Asp Tyr Lys Ala Ala Gly Gln Glu Ala
Ile Ala Lys Ala 165 170
175 Asn Pro Leu Val Val Ala Glu Lys Val Val Lys Phe 180
185
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