CN104911191B - A kind of sterility changing gene FG3 and its application - Google Patents

Abstract

The present invention relates to a kind of fertile gene and its applications, belong to biotechnology, it is prepared more particularly to plant hybridization breeding method, including sterile line propagation and hybrid seed, relates more specifically to a sterility changing gene FG3 and its mutant and its application in crossbreeding.

Description

A kind of fertility regulating gene FG3 and its application

technical field

The invention belongs to the field of biotechnology, and specifically relates to a plant hybrid breeding method, including sterile line propagation and hybrid seed preparation, and more specifically relates to a fertility gene FG3 and its mutant and its application in hybrid breeding.

technical background

Hybrid breeding is an effective way to improve crop production. Hybrids usually have obvious yield, resistance and adaptability advantages over conventional varieties. The breeding cycle of hybrids is usually shorter and quicker than conventional breeding. At present, hybrid breeding has become the main breeding method for many crops.

The selection of crop male sterile lines is a key link in hybrid breeding. The male sterile line is a plant individual with defective male gamete development and normal female gamete development, and receives pollen from the male parent as the female parent. Several factors need to be considered in the breeding of male sterile lines: 1. Hybrid matching: the combination of sterile lines and corresponding male plants produces hybrid offspring with excellent traits; Restoring fertility under certain conditions so that it can be maintained; 3. CMS self-propagation and hybrid seed production efficiency: a good sterile line must be easy to reproduce, and the yield of hybrid seed production is high. Male sterile lines currently used in rice hybrid breeding include cytoplasmic sterile lines and nuclear sterile lines. The cytoplasmic male sterile hybrid breeding system involves male sterile lines, restorer lines and maintainer lines, the so-called three-line method. Three-line hybridization requires specific restorer lines and maintainer lines, which not only complicates the seed production process, but also greatly limits the utilization of heterosis for different varieties of resources. The two-line method corresponding to the three-line method utilizes the sterile line controlled by the nuclear gene and the characteristics of the sterile line to restore fertility under specific growth and environmental conditions, so that the restorer line and the maintainer line are combined into one. Compared with the "three-line method", the "two-line method" has obvious advantages. It not only saves the maintainer line and simplifies the production procedure of hybrid seeds, but also greatly expands the selection range of male parents, and can combine various excellent traits. combined into hybrid offspring.

The key characteristics of the sterile line used for two-line hybridization are: under certain conditions, the sterile line maintains sterility and can be used for hybrid seed production; and when the conditions change, the sterile line can restore its fertility and reproduce itself, so as to maintain the sterile line. CMS purpose. At present, most of the sterile lines used in the two-line hybridization are light-temperature-sensitive sterile lines, and their fertility is affected by changes in temperature and light. The instability of these environmental factors will affect the stability of the sterile lines, resulting in self-fertilization and reduced fertility. The purity of hybrid seeds is reduced, which increases the risk of seed production. And the sterile lines that can be used for two-line hybridization are very limited by the current technology, for example, there are almost no good two-line hybrid combinations in japonica rice varieties, which limits the full utilization of variety resources.

In order to solve the defects in the current rice hybrid breeding methods, such as the stability of the sterile line, the limitation of hybrid variety resources, the complexity of seed production technology, the high cost of seed production and other technical bottlenecks, people are trying new hybrid breeding techniques, new The hybrid breeding technology will make full use of the male sterility gene controlled by the recessive nuclear gene, construct a sterile line with stable fertility and not affected by the environment, remove the constraints of environmental factors on hybrid breeding, and eliminate potential risks in production; at the same time, the The recessive sterility gene used should be applicable to the vast majority of varieties, so that the utilization of heterosis resources can be greatly improved, and the problem of resource utilization of heterosis can be solved; The male sterile line of the male sterile line has stable fertility, is not affected by environmental conditions, and can be restored by wild-type transgene. The gene and the sterile line produced by the gene mutation provide the necessary elements for the construction of a new hybrid breeding system.

Brief description of the invention

The present invention provides a DNA sequence, which has the function of regulating plant fertility, and is characterized in that the DNA sequence is selected from one of the following groups of sequences:

a) have the nucleotide sequence shown in SEQ ID NO: 1 or 5;

b) have the nucleotide sequence shown in SEQ ID NO: 10 or 11;

c) have the nucleotide sequence shown in SEQ ID NO: 13 or 14;

d) have the nucleotide sequence shown in SEQ ID NO: 16 or 17;

e) have the nucleotide sequence shown in SEQ ID NO: 19;

f) have the nucleotide sequence shown in SEQ ID NO: 21 or 22;

g) a DNA sequence capable of hybridizing to the DNA of any of the sequences described in (a)-(f) under stringent conditions; or

h) A DNA sequence complementary to any one of (a)-(g).

The amino acid sequence encoded by the above DNA sequence is shown in SEQ ID NO: 2, 6, 12, 15, 18, 20 or 23.

The present invention also provides an expression cassette, which is characterized in that the expression cassette comprises the above-mentioned DNA sequence.

The present invention also provides an expression vector, which is characterized in that the expression vector comprises the above-mentioned expression cassette.

The present invention also provides an engineering bacterium, which is characterized in that the engineering bacterium contains the above-mentioned expression vector.

The present invention also provides an application of a gene in plant fertility regulation, characterized in that the nucleotide sequence of the fertility regulation gene is selected from one of the following groups of sequences:

a) have the nucleotide sequence shown in SEQ ID NO: 1 or 5;

b) have the nucleotide sequence shown in SEQ ID NO: 10 or 11;

c) have the nucleotide sequence shown in SEQ ID NO: 13 or 14;

d) have the nucleotide sequence shown in SEQ ID NO: 16 or 17;

e) have the nucleotide sequence shown in SEQ ID NO: 19;

f) have the nucleotide sequence shown in SEQ ID NO: 21 or 22;

g) a DNA sequence capable of hybridizing to the DNA of any of the sequences described in (a)-(f) under stringent conditions; or

h) A DNA sequence complementary to any one of (a)-(g).

The present invention also includes a method for obtaining male sterile material by mutating the fertility regulating gene SEQ ID NO: 1, 5, 10, 11, 13, 14, 16, 17, 19, 21 or 22.

The "mutation" mentioned in the present invention includes substitution, deletion or addition of one or more nucleotides in the nucleotide sequence of the fertility regulation gene.

The present invention also provides a method for restoring male sterility caused by mutations in genes shown in corresponding SEQ ID NO: 1, 5, 10, 11, 13, 14, 16, 17, 19, 21 or 22, so that male sterility Fertile mutants are restored to fertile methods.

The present invention also includes the application of a mutant material, characterized in that the mutant material is caused by a mutation of a nucleotide sequence, such as SEQ ID NO: 1, 5, 10, 11, 13 , 14, 16, 17, 19, 21 or 22.

The above-mentioned "mutation" may be a point mutation, or a DNA deletion or insertion mutation, or may be generated by gene silencing methods such as RNAi and site-directed mutation.

The present invention also provides a method for applying the above-mentioned materials and DNA sequences to breeding, more specifically, the application refers to using the mutant plants as the female parent of the sterile line and crossing with the restorer line to produce hybrid seeds.

The present invention also provides a promoter, which has anther-specific expression characteristics, and its nucleotide sequence is shown in SEQ ID NO: 3 or 9. The present invention also includes expression cassettes and vectors comprising the above-mentioned promoter and/or engineered strains.

The present invention also provides a method for expressing a target nucleotide sequence in a plant, the method comprising introducing a DNA construct into a plant, the DNA construct containing a promoter and a target sequence operably linked to the promoter Nucleotide sequence, wherein the nucleotide sequence of the promoter is shown in SEQ ID NO: 3 or 9.

The "target nucleotide sequence" can be a structural gene, a regulatory gene, an antisense gene of a structural gene, an antisense gene of a regulatory gene, or a small RNA that can interfere with the expression of an endogenous gene, and its specificity in the late stage of pollen development Expression can regulate pollen fertility and pollen germination.

The present invention also includes the application of the above-mentioned DNA sequence and or promoter in any of the following (a) to (d):

(a) cultivating plant varieties or strains;

(b) Breeding plant varieties or lines with enhanced pollination and fertilization capabilities;

(c) Breeding plant varieties or strains with reduced ability to pollinate and fertilize;

(d) Breeding male sterile plant varieties or lines.

The present invention also provides a method for maintaining the homozygous recessive state of male sterile plants (for detailed methods, refer to PCT patent PCT/CN2013/086657), the method comprising:

a) providing a first plant comprising a homozygous recessive allele of the FG3 gene and which is male sterile;

b) introducing a construct into a first plant to form a second plant comprising a homozygous recessive allele of the FG3 gene and the construct, and the construct is half in the second plant Combined state, the construct comprises:

i) a first nucleotide sequence comprising a FG3 nucleotide sequence which, when expressed in the first plant, will restore male fertility;

ii) a second nucleotide sequence which, when expressed, inhibits the formation or function of fertile male gametes in said second plant, in particular the pollen inactivation gene ZM-PA; and

c) fertilizing said first plant with the male gametes of said second plant to produce offspring that retain the homozygous recessive state of said first plant.

Description of drawings

Figure 1 Morphology of florets of Osfg3 mutant and wild type Huanghuazhan.

Fig. 2 Anther morphology of Osfg3 mutant and wild type Huanghuazhan.

Fig. 3 Pollen staining analysis of Osfg3 mutant and wild type Huanghuazhan.

Figure 4 Comparison of female organ morphology between Osfg3 mutant and wild type Huanghuazhan.

Figure 5 The exposed stigma of the Osfg3 mutant, the arrow points to the exposed stigma.

Fig. 6 is a comparison of the leaf trichomes of the Osfg3 mutant and the wild type Huanghuazhan, and the arrows point to the leaf trichomes.

Fig. 7 is a comparison of the lemma trichomes of the Osfg3 mutant and the wild type Huanghuazhan, and the arrow points to the lemma trichomes.

Figure 8 shows the cDNA sequence alignment results of Huanghuazhan OsFG3, Osfg3 mutant and Nipponbare Osfg3, where HHZ indicates the sequence of Huanghuazhan OsFG3, Mutant indicates the sequence of the Osfg3 mutant, and Nip indicates the sequence of Osfg3 in Nipponbare.

Figure 9 shows the protein sequence alignment results of Huanghuazhan OsFG3, Osfg3 mutant and Nipponbare Osfg3, where HHZ indicates the sequence of Huanghuazhan Osfg3, Mutant indicates the sequence of the Osfg3 mutant, and Nip indicates the sequence of OsFG3 in Nipponbare.

Figure 10 Analysis of the expression level of OsFG3 in different rice tissues and organs.

Figure 11 Analysis of the differential expression of the OsFG3 gene between the mutant and the wild type.

Figure 12 The promoter expression vector of OsFG3 gene.

Figure 13 Transgenic functional complementation vector of rice male sterile mutant (Osfg3).

Fig. 14 RNAi interference vector of OsFG3 gene.

Figure 15 The protein encoded by the OsFG3 gene is compared with the homologous protein in the genomes of barley (Hordeum vulgare), sorghum (Sorghumbicolor), millet (millet) (Setaria italica), two spike grass (Brachypodium distachyon), corn (Zea mays) Sequence alignment diagram, 1 is rice (Oryza sativa), 2 is barley (Hordeum vulgare), 3 is sorghum (Sorghum bicolor), 4 is millet (millet) (Setaria italica), 5 is Brachypodium distachyon ), 6 is corn (Zea mays).

Detailed description of the invention

All references mentioned herein are hereby incorporated by reference.

Unless defined to the contrary, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Unless indicated to the contrary, the techniques used or referred to herein are standard techniques well known to those of ordinary skill in the art. The materials, methods, and examples are illustrative only and not limiting.

The invention includes a fertility-related gene and its nucleotide and protein sequences, and also includes the application of the gene in regulating the male fertility of plants through manipulation. By way of non-limiting example, any method described below can be used together with the corresponding nucleotide sequence provided by the present invention, for example, introducing the mutant sequence of the fertility gene into a plant to cause male sterility in the plant, Mutate the endogenous sequence of the plant, introduce the antisense sequence of the sequence into the plant, use the hairpin form, or link it with other nucleotide sequences to regulate the phenotype of the plant, or those known by those skilled in the art can be used Any of a variety of methods of affecting the male fertility of a plant.

The fertility gene FG3 provided by the present invention is a gene related to pollen development. In rice, the fertility gene is located on chromosome 10, its nucleotide sequence in indica rice is shown in SEQ ID NO: 1 or 4, and its amino acid sequence is shown in SEQ ID NO: 2; its nucleotide sequence in japonica rice The acid sequence is shown in SEQ ID NO:5, and its amino acid sequence is shown in SEQ ID NO:6; The nucleotide sequence of the fertility gene in barley is shown in SEQ ID NO:10 or 11, and its amino acid sequence is shown in Shown in SEQ ID NO:12; The nucleotide sequence of the fertility gene in sorghum is shown in SEQ ID NO:13 or 14, and its amino acid sequence is shown in SEQ ID NO:15; The fertility gene ZmFG3 in maize The nucleotide sequence is shown in SEQ ID NO: 16 or 17, and its amino acid sequence is shown in SEQ ID NO: 18; The nucleotide sequence of the fertility gene in millet is shown in SEQ ID NO: 19, and its The amino acid sequence is shown in SEQ ID NO:20; the nucleotide sequence of the fertility gene in Brachypodium distachyon is shown in SEQ ID NO:21 or 22, and its amino acid sequence is shown in SEQ ID NO:23.

The present invention also includes one of the sequences of the following groups: a) a DNA sequence having at least 90% (preferably at least 95%) sequence similarity with the above-mentioned FG3 gene sequence and having the same function; b) under stringent conditions, it can be compared with ( a) a DNA sequence of DNA hybridization of said sequence; c) a DNA sequence complementary to any of the above-mentioned sequences.

The aforementioned fertility genes can be isolated from various plants. Those skilled in the art should know that the fertility restoration gene of the present invention includes a highly homologous functional equivalent body sequence with the FG3 gene and having the same fertility regulation function. The highly homologous functional equivalent body sequence includes a DNA sequence capable of hybridizing with the nucleotide sequence of the FG3 gene disclosed in the present invention under stringent conditions. The "stringent conditions" used in the present invention are well known, including, for example, hybridization at 60° C. for 12-16 hours in a hybridization solution containing 400 mM NaCl, 40 mM PIPES (pH 6.4) and 1 mM EDTA, and then at 65° C. Wash with washing solution containing 0.1 SDS and 0.1% SSC for 15-60 minutes.

Functional equivalent sequences also include those that have at least 90%, 95%, 96%, 97%, 98%, or 99% sequence similarity with the sequence shown by the FG3 gene disclosed in the present invention, and have fertility regulation function A DNA sequence that can be isolated from any plant. Wherein, the percentage of sequence similarity can be obtained by known bioinformatics algorithms, including Myers and Miller algorithm (Bioinformatics, 4 (1): 11-17, 1988), Needleman-Wunsch global comparison method (J.Mol. Biol., 48 (3): 443-53, 1970), Smith-Waterman local comparison method (J.Mol.Biol., 147: 195-197, 1981), Pearson and Lipman similarity search method (PNAS, 85 (8): 2444-2448, 1988), the algorithm of Karlin and Altschul (Altschul et al., J. Mol. Biol., 215(3): 403-410, 1990; PNAS, 90: 5873-5877, 1993). This is familiar to those skilled in the art.

The gene sequence described in the present invention can be isolated from any plant, including but not limited to Brassica, maize, wheat, sorghum, Sorghum, white mustard, castor bean, sesame, cottonseed, linseed, soybean, Arabidopsis, Phaseolus, Peanut, Alfalfa, Oats, Rapeseed, Barley, Oats, Rye, Millet, Milo, Triticale, Einkorn, Spelt, Emmer , flax, Gramma grass, rubbing grass, false sorghum, fescue, perennial wheatgrass, sugar cane, cranberry moss, papaya, banana, safflower, oil palm, melon, apple, cucumber, dendrobium, Gladiolus, chrysanthemum, liliaceae, cotton, eucalyptus, sunflower, brassica, sugar beet, coffee, ornamental plants and pines, etc. Preferably, plants include corn, soybean, safflower, mustard, wheat, barley, rye, rice, cotton and sorghum.

The invention also provides a method for affecting plant fertility by affecting the nucleotide sequence of FG3 or regulating the transcription and expression of FG3 gene. The affecting the fertility of the plant refers to changing the fertility of the plant by regulating the expression of the FG3 gene, such as causing male sterility in the plant. Specifically, depending on specific application requirements, various methods can be used to influence the expression of the FG3 gene in plants, so as to achieve the effect of regulating the male fertility of plants. More specifically, regulating the expression of the FG3 gene can be carried out using many tools available to those of ordinary skill in the art, for example, through mutation, mutagenesis, introduction of antisense genes, co-suppression or introduction of hairpin structures, etc., can be It is used to destroy the normal expression of FG3 gene to obtain male sterile plants. On the other hand, the present invention also includes restoring the male fertility of plants whose expression of FG3 has been disrupted by introducing the nucleotide sequence of wild-type FG3 into the plants.

The invention also provides a sterile mutant sequence of the FG3 gene and the material of the male sterile mutant. More specifically, the male sterile mutant material is a process in which the plant body loses male fertility by mutating the rice endogenous FG3 gene, or mutating the nucleotide sequence of a gene highly homologous to it. The "mutation" includes but is not limited to the following methods, such as gene mutation caused by physical or chemical methods, chemical methods include mutagenesis caused by treatment with mutagens such as EMS, and the mutation can also be a point mutation, or It can be DNA deletion or insertion mutation, and can also be generated by gene silencing methods such as RNAi and site-directed mutation.

Specifically, the present invention also provides a rice male sterile mutant, which contains a mutated male sterile gene, the nucleotide sequence of the mutated male sterile gene is shown in SEQ ID NO:7, The amino acid sequence is shown in SEQ ID NO:8. Compared with the wild type, in the sterile mutant, the splicing sites of the eighth and ninth exons of the gene were mutated from GT...AG to AT...AG (Figure 8), so that the corresponding introns could not is spliced out (Figure 8), resulting in premature termination of the gene. . Those skilled in the art should know that the nucleotide sequence of SEQ ID NO: 7 can be constructed into a plant expression vector for plant transformation to obtain new transgenic male sterile mutant materials.

The present invention also provides a promoter of FG3 gene, said promoter has the function of specific expression in pollen, and the sequence of the corresponding promoter is the nucleotide sequence of about 700bp to 2500bp from ATG to upstream of FG3 gene. More specifically, in rice, the nucleotide sequence of the OsFG3 gene promoter is shown in SEQ ID NO:3 or SEQ ID NO:9. Connect SEQ ID NO:3 or SEQ ID NO:9 to the reporter gene GUS, transform the plants, detect and analyze the GUS expression activity and expression pattern in the transgenic plants, and perform GUS in the roots, stems, leaves and flowers of the transgenic plants Staining analysis showed that the OsFG3 promoter-driven GUS gene was mainly expressed in rice anthers, more specifically, it was highly expressed specifically in the P9 stage of anther development. It shows that the promoter of SEQ ID NO:3 or SEQ ID NO:9 provided by the present invention is an anther-specific expression promoter.

The plant pollen-specific expression promoter provided by the present invention contains the nucleotide sequence shown in SEQ ID NO: 3 or SEQ ID NO: 9 in the sequence listing, or contains the nucleotide sequence shown in SEQ ID NO: 3 or SEQ ID NO: 9. The sequence of nucleotide sequences has more than 90% similarity of nucleotide sequences, or contains 100 and more than 100 continuous nucleotide fragments derived from the sequence of SEQ ID NO:3 or SEQ ID NO:9, and can drive Expression of the nucleotide sequence operably linked to the promoter in plant pollen. Expression vectors, transgenic cell lines and host bacteria containing the above sequences all belong to the protection scope of the present invention. A pair of primers for amplifying any nucleotide fragment of the promoter of SEQ ID NO:3 or SEQ ID NO:9 disclosed in the present invention is also within the protection scope of the present invention.

The promoter nucleotide sequence provided by the present invention can also be used to isolate corresponding sequences from other plants than rice, especially for homologous cloning from other monocotyledonous plants. Based on the sequence homology between these corresponding sequences and the promoter sequences listed herein, or the homology with this promoter gene, techniques such as PCR and hybridization are used to identify and isolate these corresponding sequences. Accordingly, corresponding fragments isolated based on their sequence similarity to the promoter sequences of SEQ ID NO: 3 or SEQ ID NO: 9 (or fragments thereof) listed in the present invention are also included in the embodiments.

The "promoter" in the present invention refers to a DNA regulatory region, which usually includes a TATA box that can direct RNA polymerase II to initiate RNA synthesis at a suitable transcription initiation site of a specific coding sequence. The promoter may also contain other recognition sequences, usually located upstream or 5' of the TATA box, often referred to as upstream promoter elements, which function to regulate transcription efficiency. Those skilled in the art should know that although the nucleotide sequence for the promoter region disclosed in the present invention has been identified, other regulatory elements that are isolated and identified in the upstream region of the TATA box of the specific promoter region identified in the present invention are also included in the present invention. within the scope of the invention. Accordingly, the promoter regions disclosed herein are generally further defined to include upstream regulatory elements, such as those used to regulate the tissue and temporal expression function of the coding sequence, enhancers, and the like. In the same manner, promoter elements enabling expression in tissues of interest (eg, male tissues) can be identified, isolated and used with other core promoters to verify male-tissue-preferred expression. A core promoter refers to the minimal sequence required to initiate transcription, such as a sequence known as a TATA box, which is commonly found in promoters of genes encoding proteins. Thus, alternatively, the upstream promoter of the FG3 gene can be used in association with its own core promoter or from other sources.

The core promoter can be any known core promoter, such as cauliflower mosaic virus 35S or 19S promoter (US Patent No. 5,352,605), ubiquitin promoter (US Patent No. 5,510,474), IN2 core promoter ( US Patent No. 5,364,780) or the Scrophulariaceae mosaic virus promoter.

The function of the gene promoter can be analyzed by the following method: the promoter sequence is operably connected with the reporter gene to form a transformable construct, and then the construct is transferred into the plant, and in obtaining the transgenic offspring, by Observe the expression of the reporter gene in various tissues and organs of the plant to confirm its expression characteristics; or subclone the above construct into the expression vector for transient expression experiments, and detect the function of the promoter or its regulatory region through transient expression experiments

Selection of an appropriate expression vector for testing the function of a promoter or regulatory region will depend on the host and the method of introducing the expression vector into the host, such methods are well known to those of ordinary skill in the art. For eukaryotes, regions in the vector include regions that control transcription initiation and control processing. These regions are operably linked to reporter genes including YFP, UidA, GUS gene or luciferase. Expression vectors containing putative regulatory regions located in genomic fragments can be introduced into intact tissues, such as staged pollen, or into callus tissue for functional validation.

In addition, the promoter of the present invention can be linked to a nucleotide sequence other than the FG3 gene to express other heterologous nucleotide sequences. The promoter nucleotide sequence of the present invention and fragments and variants thereof can be assembled together with heterologous nucleotide sequences in an expression cassette for expression in a plant of interest, more specifically, in the male organ of the plant Express. The expression cassette has suitable restriction sites for insertion of the promoter and heterologous nucleotide sequence. These expression cassettes can be used to genetically manipulate any plant to obtain the desired corresponding phenotype.

The FG3 promoter disclosed in the present invention, more specifically the FG3 promoter in rice, can be used to drive the expression of the following heterologous nucleotide sequences, so that transformed plants can obtain male sterile phenotypes. The heterologous nucleotide sequence can encode enzymes or modifying enzymes, amylases, debranching enzymes and pectinases that promote carbohydrate degradation, more specifically such as alpha amylase genes, auxin (auxin), rot B, cell Toxin gene, diphtheria toxin, DAM methylase, avidin, or can be selected from prokaryotic regulatory system, and can also be a dominant male sterility gene.

In some embodiments, the nucleic acid mentioned in the present invention is operably linked downstream of the promoter of the present invention, wherein the "nucleic acid" can be a structure operably linked to the promoter disclosed herein genes, regulatory genes, antisense genes to structural genes, antisense genes to regulatory genes, or small RNAs capable of interfering with the expression of endogenous genes.

More specifically, the fertility regulation genes SEQ ID NO: 1 and SEQ ID NO: 5 provided by the present invention can be constructed downstream of the promoter SEQ ID NO: 3 or SEQ ID NO: 9, thereby driving the fertility regulation The specific expression of the gene in pollen, or through the technical principle of RNAi, construct the RNAi vector that can silence the gene of SEQ ID NO:1 initiated by SEQ ID NO:3 or SEQ ID NO:9, so as to obtain the gene of SEQ ID NO:1 male sterile mutants.

The promoter sequence provided by the present invention can be isolated from any plant, including but not limited to Brassica, maize, wheat, sorghum, genus Aurina, white mustard, castor bean, sesame, cottonseed, linseed, soybean, Arabidopsis Brassica, Phaseolus, Peanut, Alfalfa, Oats, Rapeseed, Barley, Oats, Rye, Millet, Milo, Triticale, Einkorn, Spelt, Emmer, Flax, Gramma grass, Fried Grass, Milo, Fescue, Perennial Wheatgrass, Sugar Cane, Cranberry, Papaya, Banana, Safflower, Oil Palm, Cantaloupe, Apple, Cucumber, Dendrobium, Sword Orchid, chrysanthemum, liliaceae, cotton, eucalyptus, sunflower, brassica, sugar beet, coffee, ornamental plants and pines, etc. Preferably, plants include corn, soybean, safflower, mustard, wheat, barley, rye, rice, cotton and sorghum.

The present invention also includes constructs containing the FG3 gene and/or its promoter, said constructs comprising so-called vectors or expression cassettes. The promoter in the construct may be the native promoter or a substituted promoter which will drive the expression of the attached nucleotide sequence in the plant. The promoter in the construct may be an inducible promoter. When the nucleotide sequence of the FG3 gene is connected with another promoter, preferably, the promoter can fully drive the expression of the sequence in the early stage of pollen development, for example, it can be specifically expressed in the P9 stage of pollen development. Specifically, the types of promoters that can be used include constitutive viral promoters, such as cauliflower mosaic virus (CaMV) 19S and 35S promoters, or Scrophulariaceae mosaic virus 35S promoter, or ubiquitin promoters.

Tissue-specific expression promoters can be used to target specific plant tissues for enhanced transcription and/or expression. A promoter may be expressed in the target tissue as well as in other plant tissues, may be strongly expressed in the target tissue and to a much lesser extent than other tissues, or may be highly preferentially expressed in the target tissue. In one embodiment, the promoter is of a type that prefers expression specifically in male or female tissues of the plant. The present invention does not require any particular male tissue-preferred promoter to be used in the methods, any of the many such promoters known to those skilled in the art may be used. The native FG3 promoter described herein is one example of a promoter that can be used. Another such promoter is the 5126 promoter, the MS45 promoter, the MS26 promoter, the BS92-7 promoter, the SGB6 regulatory element, and the TA29 promoter, among others, which preferentially direct the expression of the gene to which it is linked in male plant tissues . A gamete tissue-preferred expression promoter may also be included in some constructs. Male gametes preferentially express promoters including PG47 promoter and ZM13 promoter.

The above-mentioned constructs may also include other components, which mainly depend on the purpose and use of vector construction, for example, may further include selectable marker genes, targeting or regulatory sequences, stabilizing sequences or guide sequences, introns, and the like. The expression cassette will also include a transcriptional and translational terminator functional in plants at the 3' end of the heterologous nucleotide sequence of interest. The terminator can be the terminator of the gene provided by the present invention, or a terminator from an external source. More specifically, the above terminator may be a nopaline synthase or octopine synthase termination region.

When it is desired to direct the expression product of a heterologous nucleotide sequence to a specific organelle, such as a plastid, amyloplast, or to the endoplasmic reticulum, or to secrete it on the cell surface or extracellularly, the expression cassette may also contain a Nucleotide sequence encoding the transit peptide. Such transit peptides are well known in the art, and include but not limited to the small subunit of Rubisco, plant EPSP synthase, maize Brittle-1 chloroplast transit peptide, and the like.

In preparing the expression cassette, various DNA fragments can be manipulated to provide the DNA sequence in the proper orientation, or in the correct reading frame. For this purpose, adapters or adapters may be used to connect the DNA fragments, or further manipulations may be included to provide convenient restriction enzyme sites and the like.

Furthermore, the construct provided by the present invention may also include a selectable marker gene for selection of transformed cells or tissues. The selectable marker genes include genes that confer antibiotic resistance or resistance to herbicides. Suitable selectable marker genes include, but are not limited to: chloramphenicol resistance gene, hygromycin resistance gene, streptomycin resistance gene, spectinomycin resistance gene, sulfonamide resistance gene, glyphosate resistance gene , the grasshopper resistance gene. The selection marker gene can also be red fluorescent gene, cyan fluorescent protein gene, yellow fluorescent protein gene, luciferase gene, green fluorescent protein gene, anthocyanin p1 and other genes.

The expression cassette or vector provided by the present invention can be inserted into a plasmid, a cosmid, a yeast artificial chromosome, a bacterial artificial chromosome or any other vector suitable for transformation into a host cell. Preferred host cells are bacterial cells, especially for cloning or storing polynucleotides, or for transforming plant cells, such as Escherichia coli, Agrobacterium tumefaciens and Agrobacterium rhizogenes. When the host cell is a plant cell, the expression cassette or vector can be inserted into the genome of the transformed plant cell. Insertion can be positional or random insertion. Preferably, insertion is achieved such as by homologous recombination. Alternatively, the expression cassette or vector can be maintained extrachromosomally. The expression cassette or vector of the invention may be present in the nucleus, chloroplast, mitochondria and/or plastid of plant cells. Preferably, the expression cassette or vector of the invention is inserted into the chromosomal DNA of the plant cell nucleus.

The present invention also includes the application of the disclosed FG3 gene and its promoter. In some application embodiments, the FG3 gene or its promoter provided by the present invention can be used to realize FG3 or other similar fertility-related gene mutations. Propagation and maintenance of acquired male sterile lines.

Specifically, the reproduction and maintenance of the above-mentioned male sterile line refers to using a homozygous recessive nuclear male sterile mutant as a transformed recipient material, and transforming three closely linked target genes into the sterile mutant recipient plant . The three target genes are fertility restoration gene, pollen inactivation gene and color marker screening gene respectively. Among them, the fertility restoration gene can restore the fertility of the sterile transformed subject, the pollen inactivation gene can inactivate the pollen containing the transformed foreign gene, that is, lose the fertilization ability, and the screening gene can be used for transgenic seeds and non-transgenic seeds Sorting, the sorted non-transgenic seeds are used as sterile lines to produce hybrids, and the transgenic seeds are used as maintainers to continuously and stably produce sterile lines.

More specifically, according to an embodiment of the present invention, the rice nuclear recessive sterile fg3/fg3 mutant can be used as the transformation recipient material, and three closely linked target genes can be transformed into the sterile line: wherein, the fertility The restorer gene OsFG3 can make the transformation subject recover; the pollen inactivation gene Zm-PA can inactivate the pollen containing the exogenous gene, that is, lose the ability of fertilization; the fluorescent color sorting gene RFP(r) is used for transgenic and non-transgenic seeds Sorting, the sorted non-transgenic seeds are used as sterile lines to produce hybrids, and the transgenic seeds are used as maintainers to continuously and stably produce sterile lines. Because this technology uses biotechnology to produce non-transgenic products, it solves the bottleneck problem faced in the process of rice hybrid seed production, that is, the low utilization rate of resources in the three-line method and the unstable fertility of the sterile line in the two-line method (detailed methods can be found in See PCT patent PCT/CN2013/086657).

The pollen-specific expression promoter provided by the present invention can be used for the specific expression of exogenous genes in pollen, thereby avoiding the adverse effects brought by the continuous expression of the exogenous gene in other tissues of plants, and can also be used for plant pollen Functional analysis and identification of growth and development-related genes; can be used to create male sterile lines and restorer lines; and can be applied to pollen abortion experiments to avoid biosafety problems caused by plant transgene drift or pollen escape. The creation of plant male sterile lines and restorer lines is of great significance.

The present invention also provides a plant production method, which includes:

(1) constructing the expression cassette provided by the second aspect or the fifth aspect of the present invention;

(2) introducing the expression cassette obtained in step (1) into plant cells;

(3) regenerate the transgenic plant; and

(4) selecting transgenic plants; and

(5) Optionally, propagating the plants obtained in step (4) to obtain progeny.

The transgenic plants of the present invention are prepared using transformation methods known to those skilled in the art of plant biotechnology. Any method can be used to transform the recombinant expression vector into plant cells to produce the transgenic plants of the invention. Transformation methods can include direct and indirect transformation methods. Suitable direct methods include polyethylene glycol-induced DNA uptake, liposome-mediated transformation, introduction using a gene gun, electroporation, and microinjection, among others. In a specific embodiment of the invention, the present invention uses Agrobacterium-based transformation technology (see Horsch RB et al. (1985) Science 225:1229; WhiteFF, Vectors for Gene Transfer in Higher Plants, Transgenic Plants, Vol. 1, Engineering and Utilization, Academic Press, 1993, pp.15-38; Jenes B et al. Techniques for Gene Transfer, Transgenic Plants, Vol. 1, Engineering and Utilization, Academic Press, 1993, pp.128-143, etc.). Agrobacterium strains (such as A. tumefaciens or A. rhizogenes) contain a plasmid (Ti or Ri plasmid) and T-DNA elements that are transferred to plants after transfection with Agrobacterium, and the T-DNA is integrated into the genome of plant cells. The T-DNA can be located on a Ri-plasmid or a Ti-plasmid, or contained independently in so-called binary vectors. Agrobacterium-mediated transformation methods are described in eg. Agrobacterium-mediated transformation is most suitable for dicots, but is also suitable for monocots. Transformation of plants with Agrobacterium is described, eg, in . Transformation can result in transient or stable transformation and expression. Although the nucleotide sequences of the present invention can be inserted into any plant and plant cell falling within these broad categories, they are especially useful in crop plant cells.

Compared with the prior art, the present invention has the following beneficial effects: the present invention provides a rice pollen development gene and a male sterile line produced based on the mutation of the gene. The fertility of the sterile line is stable and is not affected by the environment. Conditional effects, able to be restored by wild-type transgenes. The gene and the sterile line produced by the gene mutation provide the necessary elements for the construction of the third-generation hybrid breeding system. The male sterile line produced by the gene mutation is used to produce hybrid seeds. Line" and "two-line" hybridization techniques are of great significance.

Detailed ways

The embodiments of the present invention are described in detail below. This embodiment is implemented on the premise of the technical solution of the present invention, and detailed implementation methods and specific operating procedures are provided, but the protection scope of the present invention is not limited to the following implementation example.

Embodiment 1, rice male sterile mutant (osfg3) screening

The mutant was obtained by mutagenizing the indica rice Huanghuazhan seeds (M0) with EMS, the concentration and time of EMS mutagenesis were 0.7%, 12 hours, and the mutant pool (M1) was obtained from the seed plants of the M0 generation after they were fruited. Plants from M1 generation seeds were used for screening at the seed maturity stage, and sterile plants were obtained through phenotype observation. Sterile plants were regenerated by cutting rice piles, and pollen development and staining reactions of regenerated plants were detected by I2-KI staining during the reproductive period. One of the mutants exhibited no pollen sterility (no pollen) and was named osfg3.

Embodiment 2, rice male sterile mutant (osfg3) genetic analysis

The sterile plants of the osfg3 mutant were crossed with the wild type Huanghuazhan, and the F1 plants of the three hybrid populations were all fertile. Then the F1 generation was selfed, and the segregation ratio of sterile and fertile plants in the F2 generation was close to 1:3 (Table 1), indicating that the mutation was controlled by a recessive single gene.

Table 1: Segregation ratio of F2 generation of rice male sterile mutant (osfg3) hybrid

Embodiment 3, rice male sterile mutant (osfg3) reproductive organ phenotype analysis

Compared with the wild type, the mutant plants grew and developed normally, and flowered at the same time. The size, shape, opening size and opening time of palea and palea were not different from wild type (Fig. 1). However, the mutant anthers are thin and small, white (Fig. 2), the anthers do not dehisce, and there is no pollen. Further, the pollen of the mutant is stained and detected with I2-KI solution. As shown in Fig. 3, the pollen staining of the wild type is normal, while Mutants have no pollen. The female organs (including ovary, style, and stigma) of the mutant were slightly larger than those of the wild type (Fig. 4). The stigma exposure rate of the mutant was over 85% (Fig. 5), while the stigma of the wild type Huanghua was rarely exposed. Compared with the wild type Huanghuazhan, the trichomes on the leaves of the mutant became more and smaller (Fig. 6). Compared with the trichomes of the lemma, compared with the wild type Huanghuazhan, the trichomes on the mutant lemma became more and smaller (Fig. 7).

Embodiment 4, the cloning of rice male sterile mutant gene

Mutmap method was adopted for mutant gene cloning, that is, the F2 population was constructed by crossing mutants with original wild parents, and gene positioning was carried out by resequencing. The sterile plants were crossed with the wild-type Huanghuazhan, and 30 F2 generation sterile plants were selected, and the leaves were taken to extract genomic DNA, which was mixed in equal amounts and used for high-throughput genome sequencing. A total of about 18.5Gb of genome sequence data was obtained, which is equivalent to 43x rice Genome (Table 2). Compared with the genome sequence of wild type Huanghuazhan, the mutant gene may be the Os10g35180 allele on chromosome 10.

Table 2 Resequencing data of rice male sterile mutants

The coding region of the gene in the wild type Huanghuazhan is 2181bp in length, and its nucleotide sequence is shown in SEQ ID NO:1. The protein encoded by SEQ ID NO:1 contains 727 amino acids, and its amino acid sequence is shown in SEQ ID NO:2. In the sterile mutant, the splicing sites of the eighth and ninth exons of the gene were mutated from GT...AG to AT...AG (Fig. 8), so that the corresponding introns could not be spliced out (Fig. 8), leading to premature termination of the gene. Using the latest SNP (single nucleotide polymorphism) research tool HRM (High Resolution Melt, high-resolution melting) analysis, it was further verified that all pollen-free plants carried homozygous mutant sites, while fertile plants carried Homogeneous and wild-type or heterozygous loci (Table 3). The selfed offspring of the pure and wild-type plants at this site are all fertile, and the selfed offspring of the heterozygous plants at this site are sterile: fertile 1:3 segregation.

Table 3 HRM typing results of the mutant gene (Os10g35180)

There is a nucleotide sequence polymorphism in the cDNA coding region of the gene between the japonica rice Nipponbare and the wild type Huanghuazhan (Figure 8). Mutation to C, C mutation at position 1371 to T, A mutation at position 1422 to G, G mutation at position 1515 to A, a fragment deletion at position 2298-2307, the specific missing fragment is CGCCCGCGG, and The G at position 2324 was mutated to T, resulting in a polymorphism of 3 amino acid residues at positions 696-698 of the corresponding amino acid sequence of the gene, and the amino acid residue at position 705 was mutated from alanine A to serine S (FIG. 9), specifically in the japonica rice Nipponbare, the nucleotide sequence of the gene is shown in SEQ ID NO:5, and the encoded amino acid sequence is shown in SEQ ID NO:6. Further analysis showed that there was no polymorphism in this gene between the indica variety 9311 and the wild type Huanghuazhan.

Embodiment 5, the expression analysis of OsFG3 gene in each organ of rice

Primers were designed according to the cDNA sequence of OsFG3, the upstream primer was F15'GGCTGATCACCTACTACAAGAACTC3'(SEQ ID NO:24), the downstream primer was R15'CTCCTGCATCCAGCACTGTC 3'(SEQ ID NO:25), and the rice Actin gene was used as an internal control to design primers, The upstream primer is 5'GCTATGTACGTCGCCATCCA' (SEQ ID NO:26), and the downstream primer is 5'GGACAGTGTGGCTGACACCAT' (SEQ ID NO:27). Huanghuazhan rice material was used to extract total RNA and synthesize cDNA template. Real-time quantitative PCR method was used to analyze the OsFG3 gene in rice root, stem, leaf, lemma, palea, glume, pistil and young panicle primordia differentiation stage (stage6), young panicle pollen mother cell meiosis stage ( stage7) and tetrad formation stage (stage8), microspore early stage (stage9), microspore middle and late stage (stage10) and mature pollen stage (stage12) expression profiles, the results are shown in Figure 10, the gene in microspore It is specifically expressed in the early stage (stage9) and the expression level is very high, the expression level begins to decrease in the middle and late stages of microspores (stage10), and the expression level is very low in the developmental stages of roots, stems, leaves, seeds and other young spikes. The expression of this gene was significantly down-regulated in pistil and young spike pollen mother cell meiosis stage (stage7), tetrad formation stage (stage8), early microspore (stage9) and late microspore stage (stage10) Trend (Figure 11).

Embodiment 6, construction and functional analysis of the promoter expression vector of OsFG3 gene

The construction of the promoter expression vector of OsFG3 gene (Figure 12): using primers OsFG3-Pro-F (ACTGGTACCCTTAATCCCTTCTATACC, SEQ ID NO:28) and OsFG3-Pro-R (TCTGTCGACTCATCTCTCCCTCTCTCGC, SEQ ID NO:29) to amplify the rice genome to obtain The 2010bp OsFG3 gene promoter fragment (SEQ ID NO: 3), the fragment was digested with Kpn1 and Sal 1 and then ligated into the promoter detection vector pHPG to obtain the pOsfg3-gus vector. The wild-type rice callus is transformed into the wild-type rice callus by the method mediated by Agrobacterium, and the transgenic plants are obtained through selection and regeneration.

The regenerated transgenic rice plants were screened, the expression pattern of the OsFG3 promoter was analyzed by analyzing the activity of B-galactosidase, and the roots, stems, leaves and flowers of the transgenic plants were analyzed by GUS staining, and it was found that the OsFG3 promoter drives GUS is mainly expressed in rice pollen. In addition, when the promoter SEQ ID NO:9 was connected to GUS for functional verification, it was found that the staining results were consistent with the results of SEQ ID NO:3, both of which were anther-specific expression promoters.

Example 7, Transgenic function complementation of rice male sterile mutant (osfg3)

Construction of complementary vector (Figure 13): use primers Osfg3-Res-F (ACTGGTACCCTTAATCCCTTCTATACC, SEQ ID NO:30) and Osfg3-Res-R (AGTGGATCCGGATCATGGAAATATCAAGTAC, SEQ ID NO:31) to amplify the rice genome to obtain the start codon containing Osfg3 The 2010bp base upstream of the sub-ATG and the 748-base full-length sequence fragment (SEQ ID NO: 4) after the stop codon TGA, the fragment is double-digested with Kpn1 and BamHI and then ligated into the genome complementary expression vector The pOsfg3-complementation vector was obtained in . The seed-induced callus of Huanghuazhan osfg3 mutant was transformed by Agrobacterium-mediated method, and transgenic plants were obtained by screening and regeneration.

All obtained transgene-positive plants were analyzed, and it was found that all transgene-positive plants were fertile. These analyzes further demonstrated that the gene OsFG3 is involved in the regulation of pollen development, and mutations in this gene lead to the pollenless phenotype.

Example 8, Obtaining and Phenotype Analysis of Transgenic Lines Interfering with RNAi of OsFG3 Gene

The construction of the RNAi interference vector (Figure 14) of the OsFG3 gene: use primers Osfg3-Flag-F1 (ctcagatctGATCCAAGTGGCTGGACCTC, SEQ ID NO:32) and Osfg3-Flag-R1 (ctcactagtCAGCTGCTGATATACACCTTGA, SEQ ID NO:33) to amplify rice cDNA to obtain 244bp The OsFG3 gene fragments were amplified with two pairs of amplification primers with Bgl2 and Spe1 restriction sites respectively to obtain forward and reverse OsFG3 gene fragment 1 (SEQ ID NO: 34) and OsFG3 gene fragment 2 (SEQ ID NO: 35), digested and ligated into pRNAi vector to obtain pOsfg3-RNAi 1. At the same time, another RNAi vector was constructed, using primers Osfg3-Flag-F2 (ctcagatctGATCCAAGTGGCTGGACCTC, SEQ ID NO:36), Osfg3-Flag-R2 (ctcactagtCAGCTGCTGATATACACCTTGA, SEQ ID NO:37) to amplify rice cDNA to obtain a 480bp OsFG3 gene fragment , using two pairs of amplification primers with Bgl2 and Spe1 restriction sites to amplify forward and reverse OsFG3 gene fragment 3 (SEQ ID NO: 38) and OsFG3 gene fragment 4 (SEQ ID NO: 39), the enzyme Cut into the pRNAi vector to obtain pOsfg3-RNAi2. The callus of Nipponbare was transformed by Agrobacterium-mediated method, and the transgenic plants were obtained by screening and regeneration.

Example 9, the sequence alignment of the protein encoded by the OsFG3 gene and the homologous protein predicted in the genomes of barley, sorghum, and maize

In the NCBI database, using the protein blast tool, the complete sequence of the protein encoded by the rice OsFG3 gene was searched in the protein database, and barley (Hordeum vulgare), sorghum (Sorghum bicolor), millet (millet) (Setaria italica), The homologous proteins predicted in the genomes of Brachypodium distachyon and Zea mays were compared and analyzed. The results showed that the homologous proteins from different plants had very similar conserved sequences, and The homology among them is very high (Fig. 15), indicating that the biological function of this protein is conserved during the development of male organs of plant flowers and plays a very important role.

Wherein, the nucleotide sequence of the fertility gene in barley is shown in SEQ ID NO: 10 or 11, and its amino acid sequence is shown in SEQ ID NO: 12; the nucleotide sequence of the fertility gene in sorghum is shown in Shown in SEQ ID NO:13 or 14, its amino acid sequence is shown in SEQ ID NO:15; The nucleotide sequence of this fertility gene in corn is shown in SEQ ID NO:16 or 17, and its amino acid sequence is shown in SEQ ID NO:17 Shown in ID NO: 18; The nucleotide sequence of the fertility gene in millet is shown in SEQ ID NO: 19, and its amino acid sequence is shown in SEQ ID NO: 20; The fertility gene in Brachypodium distachyon The nucleotide sequence is shown in SEQ ID NO: 21 or 22, and the amino acid sequence is shown in SEQ ID NO: 23.

SEQUENCE LISTING

<110> Shenzhen Institute of Crop Molecular Design and Breeding

Shenzhen Xingwang Biological Seed Industry Co., Ltd.

thriving investment co., ltd.

<120> A Fertility Regulation Gene FG3 and Its Application

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ctagctggaa aaccctgggc atttctctta ttttaggata actgaaaata aaccataaat 900

gatattattc tgaaagaggg aatactctct tctgcaaatg aaaaaaaaaa actataacaa 960

gaaaccatga tcgggatagc atttgtgcag cagaaagaac ctaaaccgtg tgtggctgca 1020

ttgcatgcat gatagtgatg agataccata tggctgttca caacccaaca acttgcaatc 1080

cagtgcatgc attacatggc caagaatcca gcggttgagc cactgacatg tatattccac 1140

tttcgttatg acccgcatgt cagtggctca attgtacgta cagctaagtc agagaatttc 1200

cttcatctct accaatctat tctctctctt tggcctggtt tagtcccaaa acatcatatc 1260

gaatctttga aaacatgcat gtagcattaa atatataata aaaaaaacta attgcatagt 1320

taggcggaaa tcacgagaca aatcttttga gcctaattag tccattatta gccgtaagtg 1380

ctacagtaac ccacatacgc taatgacgga ttaattaagc tcaaaagatt cgtcttgtga 1440

ttttcaggca agttatgaaa ttagtttttt cattcgtgtc cgaaaacccc taccgatatc 1500

cagtcaaaca tccgatgtga cacccaaaca tttcttttct tgaattaaac atgccctttc 1560

tcttgtgctt gcattgcata tgcatatgca caatgcattg cttctagatc acaccaaaca 1620

gctactagcc cccccttatg acatgccatt catctccat gcatgcttgc atattgcatg 1680

catgctgctg ctgcttctac tcctcctcct ctctagagct agctaccttg accctaggtc 1740

aaacccctac acccttcatt acttctgcac ctgatctcct tctctctcta gcttctctcc 1800

ctagattttc tctctctaac ctgctcctcc attackggcc tttattcca ccacctcctc 1860

ctcttctcct ccctcatagg tggtgtttgc ttagctagct agctagctcc atcactagct 1920

ctcgcatcgc atgccacggt gagccggtgt cgccggcgtc gtacggacgt agcaagcgag 1980

agaggggagag atgaagaagc agggaggagg aggaggagta gcggcggcgg cggtggcggc 2040

ggcggggatg gtgaaggcgg agctggagga cgtggggatc aaggcggcgg gaggggcggt 2100

ggcggcgctg agcccgctga gcgagacgct gtggcgggag aaggcggcgg cggagttcct 2160

cggcgacgtg tcggcgcgcc tggcgtggcg cgacctcacc gtcaccgtcg tgctcggcgg 2220

cggcggcggc ggcggcggag gagggacgac gcaggacgtg ctccaggggc tcacggggca 2280

cgccgagccg ggcaccatca cggcgctcat gggcccctcc ggctccggca agtccaccct 2340

gctcgacgcc ctcgccggcc gcctcgccgc caacgccttc ctcgacggca ccgtcctcct 2400

caacggccgc aaggccaacc tctccttcgg cgccgcggta cgcacgcaca cctcgccgcc 2460

ggcgacccgt cgccactgtg ctgaactgct gaccgcaacg ctaagttaat cagttattag 2520

tagtagtaca atagtactaa atactagtaa atactaatca gctttgcttt tatatgggat 2580

tggattttgc ttcagaaaat ggttttctgg attaaaagct gcacattaaa ctaaagaagt 2640

actccctccg tttcacgatg taaatcattc tgtcatttcg cacattcata ttgatgttaa 2700

tgaatctaaa catatataca tcaatattaa tatgggaaaa gttagaatga tttacattgt 2760

gaaacggagg aagtagttca tttcttgaag tataactgca cacagggaga atacttgtag 2820

tacaaaagaa aaaaaaacat ggaatatgtg tgtgttcttt ttgcagtact tcattttctt 2880

gaagaagatt tatttttttt tgtgtgtgtt tgattagaat gtgtattgta tatttttttt 2940

tcctttctgt tgctttcatt acttggtctt tatctgaatc acctgaacaa ccgatttcag 3000

aatgagaaag aaatgggtca tttttttgtt gttgccacgt gccatttctt ggaaccttgc 3060

tgctttctgg cttctgcatg tgtttggaca gactgcctct ctgcagatgt aatttactgc 3120

attaataact ccctttcatg gatatagatc accatttct ggccaagatt aattggggga 3180

tctggctcaa tcaattaatt agtttctgct tattaatttt tagtaggaa aaaaaaaagc 3240

tctttcaggt atgtgtttgg taatggagtt aacactcatt cagtcatcta gcttgttggc 3300

aaggaaaact aggaacttcc ccagcagatg ggatgtggtt taccagtttt ttcagaaact 3360

atgctcatca aaaactcatg atctgttgac aatcctgatt atttgttgta catgtagcgt 3420

gtgtcgccat gttgtccaaa atcatttctt tatgacttaa caggagcttt tttaaaagaa 3480

aaaacatagt ccaaaagaag gttgttgctg atatttttca tgacttgtgc agttgtgctt 3540

catttgtttc tgtcaaaact gtcaacctaa tatgtaggaa gtagtttaac ttctgtacca 3600

atcaataact ataaagtatt atcaaggaaa aaaagagcaa aatttctgta tgtaaaaagt 3660

tctaccttgt caaaaatgat aaggtgatac ctgaagttat gtgcatatat tatcttactt 3720

cgtctgatgc agtaataatc tcaaggtaac aaaagaagtt acctccaggc tccagcttac 3780

tgaaagtttc agtaaaatat gtaatttcag gcctatgtga cccaagatga caacttgatc 3840

ggcacattga cggtgaggga gaccatctca tactcggcgc ggcttcgcct tcccgacaag 3900

atgcccatgg aagagaagcg cgcccttgtc gagggcacaa tcgtcgagat ggggctgcag 3960

gactgcgccg acacggtggt cggcaattgg cacctgaggg gaatcagtgg tggtgagaag 4020

aggagggtca gcattgccct tgagatactg atgaggccca ggctgctctt cctggatgaa 4080

ccaaccagtg gccttgacag gtaggtactg ctattatatt tatggcaaat catttgttat 4140

tacattaatt aatggatcat gcaagcatgc cgggtcggcc aaaggctggc cgaccaattt 4200

cattaagatt gggagaaaat aaagtaaaca tttagttaca tccaggccca gcatgatagc 4260

aatctagcta tctagcaagc atgatagcaa tctagctgaa cttgagatgt ggagatttga 4320

tttgcacagc tctctacctt gctttgcttt caaccagtag tgtttatcaa actcattcct 4380

ggtataacta tcagaaggtt actatgcctt gtcatttgtc aagtgattgt aattgttaca 4440

gtagcaatta tctgtctaag taactgatgt gcagatgatt aatctgttgc agtgcttcag 4500

cgttcttcgt gacacagaca ctgcgtgggc tagcgaggga tggcaggaca gtgatagcat 4560

cgatccatca gccgagcagc gaggtttttg agcttttcga ccgcttgtat ctcctttcag 4620

gaggcaaaac tgtttacttt gggctggcct ctgaagcttg tcaggtatgt cagttttcac 4680

aaactctatt aagttgtttc tgcaatgcta atatctatat gggttccctc aatcatgtac 4740

tgttctttgc attttgcagt tctttgccca agccggcttc ccttgcccac cgctgcgcaa 4800

tccatcggat catttcctca gatgcataaa cgcagacttc gacaaggtga aggccactct 4860

gaaaggatca atgaagagaa gagtaagttt caggattcag tcagtctcct cctagtacct 4920

acaactggca ctcttacctg tgcatgcctt gactgaaatt gtgctcacat tttgcataca 4980

tagtttgaaa gatccgatga cccacttgat cggattatga cttcggaggc catcagaagg 5040

ctgatcacct actacaagaa ctcacagtac tactttgctg cgcagcagaa agtcaatgag 5100

atggcacggg ttgtaagtaa ctagattact acagcaactt ccccaaacat tcaaccacag 5160

ctagaccatt gtaatgcaag ctaagaggac tcacatggta tttttgttgc agaaagggac 5220

agtgctggat gcaggaggga gccaggctag cttttggatg caggctttca cactcacgaa 5280

gcgatcgttc atcaacatgt cgcgggactt cgggtactac tggctgaggc ttatcatcta 5340

cattgttgtg acagtctgca tcgggacaat ctacctcaat gttggcacca gatacagctc 5400

aattctggta aaaatttcct tcttcagttg tcacttctat ttggatgcaa acttacactg 5460

aaaagtatct agctacccctg aactgaatac ttgatgttaa gcttggtgat ctgataaaac 5520

tgtgaaaaca attaggcaag aggtgcatgc gcttctttcg tcttcggctt tgtcacgttc 5580

atgtcgattg gaggattccc atcttttgtg gaagacatga aggtttgtgt caccactgaa 5640

tcaacaaatg ggttagaaaa gttcttcaga ttacagcaaa ccatgctcac agaatctttt 5700

ttatgctctt tcaggtgttt cagagagaga ggctcaatgg gcactatggc gtgctggcat 5760

tcgtgataag caacacgatc tcggcgatgc cattcttgat cttgatcacc ttcatctccg 5820

ggacaatgtg ctacttcatg gtgcgcctcc acccgggctt cacgcactac ctcttcttcg 5880

tcctctgcct ctacgcaagc gtcaccgtcg tcgagagctt gatgatggcc attgccagtg 5940

tcatccccaa cttcctcatg ggcatcatca tcggtgcagg aattcaggtg aattaattac 6000

catccatcac aacaacttga tcctgtccat ctttatcagg cactgtttgg agtcctgcaa 6060

cgcatacaaa ctgggattaa attttgacac tgtacttgga actgcacaac ataatgttga 6120

tgtttgttaa tattctctca tgatcatgac atgcaacaac atttattgaa gagaaaaata 6180

tattttggaa atggacaggg gatattcatg ctggtttcag gatacttcag gcttccccat 6240

gacatcccaa agccattctg gaggtatccc atgtcataca tcagcttcca ctactgggca 6300

ttgcaggtaa aaatttcaac tttttttaca gctcataaac tgaagcaacc agtgaactca 6360

aatcagggtt taaaacaaca aatcaggaaa atttaaaccc aagaccaaaa aatcaaaatc 6420

aaaattttgt ccatgtgtta ctgctgttac ttctgttgtc acaagtacat gtatagaaca 6480

tgacatgaga aacaacgaat ttgagaaatt tgaagcaaaa atgggccaat ttcaatcttg 6540

acaggtttcg tcgatatttt ccataatttt tgtcaatctc ttaaactggt tatacctctg 6600

ttgtcagttt tcactagtat atgtaatact agaacataaa acaactattg ataattatac 6660

atgaggaaca ataaattcgg aaaaactacc agcaaaaaag cggcaaattt gaattttgat 6720

gggtttgatc gatatttttc ataatttttg tcaatctctt aaactgctta ttatacatct 6780

gttatcagct ttcaccacca catgtaatag aacataaaac aactatggag aattacacac 6840

gagaagcaac aaattcaagc aaaaaatgtg cagattcgaa tgtttgactg gttttttttt 6900

tccatctgac attgttgcag ggtcagtacc agaacgacct gaagggcctg gtgttcgaca 6960

accaggacga cgagctcccc aagatccccg gcgagtacat cctggagaac gtgttccaga 7020

tcgacgtgag ccgatccaag tggctggacc tcgccgtgct cttcagcatg atcttcatct 7080

accgcctcct cttcttcgcc atgatcaagg tcagcgagga cgtcaccccg tgggtgcgcg 7140

gctacgtcgc gcggcgccgg gtgcagggca agggcgcccg cggcgcccgc ggccgcgggg 7200

cggacctcgc cgccgcgcgc tcgccgtcgc tccgcgccta cgtcgtcgac gccgccgacg 7260

acctgccgcc ggcctgacca cccgtgagac agacacggct tctgaaaact ctgaaactct 7320

gaactcctca actgaacaag tggagacaga ggaagaaaaa gagagagagg gagaggctaa 7380

aactgaacta aaaaaaaaag aacctgaata tctgatatga taattgtgtg ttatagtagt 7440

agcaatgctt aattaatcga tcaagctgtg ttgttgtatt gtattgataa ttactaatat 7500

caaggtgtat atcagcagct gaatgcaagt ttagctagtg cttagtttca ggtctctgtc 7560

ttatctctga tcttgctgca gttagtagta acaacttatt caagttggaa atttgatggt 7620

tccattttgt ttggatggca tcttgcagca gtgggtttgt ccaaaatgga gctacagctg 7680

ctacagattg gcaatgatgc atatggcagc aaggtactga aagattttcac aaggatcttg 7740

tcaggacagt atgcagtttg gttgtgcaag agaaaaacat ccaagaaagg acatatgata 7800

tttatgagta ttgaagagca aactgttcgc tacttgcttg taccatgatc catcatttag 7860

tctaaacaca ttgcaagatg gtgtggcctg ttaaagaagc aattttgcaa tgttgataag 7920

taattaatga caagctgttt tatcatgtga ggagagttta tttgtatgca gacaagagca 7980

acaaactgta tatatgctag tagtacttga tattccatg atcc 8024

<210> 5

<211> 2172

<212>DNA

<213> Rice (Oryza sativa Japonica)

<400> 5

atgaagaagc agggaggagg aggaggagta gcggcggcgg cggtggcggc ggcggggatg 60

gtgaaggcgg agctggagga cgtggggatc aaggcggcgg gaggggcggt ggcggcgctg 120

agcccgctga gcgagacgct gtggcgggag aaggcggcgg cggagttcct cggcgacgtg 180

tcggcgcgcc tggcgtggcg cgacctcacc gtcaccgtcg tgctcggcgg cggcggcggc 240

ggcggcggag gagggacgac gcaggacgtg ctccaggggc tcacggggca cgccgagccg 300

ggcaccatca cggcgctcat gggcccctcc ggctccggca agtccaccct gctcgacgcc 360

ctcgccggcc gcctcgccgc caacgccttc ctcgacggca ccgtcctcct caacggccgc 420

aaggccaacc tctccttcgg cgccgcggcc tatgtgaccc aagatgacaa cttgatcggc 480

acattgacgg tgagggagac catctcatac tcggcgcggc ttcgccttcc cgacaagatg 540

cccatggaag agaagcgcgc ccttgtcgag ggcacaatcg tcgagatggg gctgcaggac 600

tgcgccgaca cggtggtcgg caattggcac ctgaggggaa tcagtggtgg tgagaagagg 660

agggtcagca ttgcccttga gatactgatg aggcccaggc tgctcttcct ggatgaacca 720

accagtggcc ttgacagtgc ttcagcgttc ttcgtgacac agacactgcg tgggctagcg 780

agggatggca ggacagtgat agcatcgatc catcagccga gcagcgaggt tttcgagctt 840

ttcgaccgct tgtatctcct ttcaggaggc aaaactgttt actttgggct ggcctctgaa 900

gcttgtcagt tctttgccca agccggcttc ccttgcccac cgctgcgcaa tccatcggat 960

catttcctca gatgcataaa cgcagacttc gacaaggtga aggccactct gaaaggatca 1020

atgaagagaa gatttgaaag atccgatgac ccacttgatc ggattatgac ttcggaggcc 1080

atcagaaggc tgatcaccta ctacaagaac tcacagtact actttgctgc gcagcagaaa 1140

gtcaatgaga tggcacgggt taaagggaca gtgctggatg caggagggag ccaggctagc 1200

ttttggatgc aggctttcac actcacgaag cgatcgttca tcaacatgtc gcgggacttc 1260

gggtactact ggctgaggct tatcatctac attgttgtga cagtctgcat cgggacaatc 1320

tacctcaatg ttggcaccag atacagctca attctggcaa gaggtgcatg tgcttctttc 1380

gtcttcggct ttgtcacgtt catgtcgatt ggaggattcc cgtcttttgt ggaagacatg 1440

aaggtgtttc agagagagag gctcaatggg cactatggcg tgctggcatt cgtgataagc 1500

aacacgatct cggcaatgcc attcttgatc ttgatcacct tcatctccgg gacaatgtgc 1560

tacttcatgg tgcgcctcca cccgggcttc acgcactacc tcttcttcgt cctctgcctc 1620

tacgcaagcg tcaccgtcgt cgagagcttg atgatggcca ttgccagtgt catccccaac 1680

ttcctcatgg gcatcatcat cggtgcagga attcaggggga tattcatgct ggtttcagga 1740

tacttcaggc ttccccatga catcccaaag ccattctgga ggtatcccat gtcatacatc 1800

agcttccact actgggcatt gcagggtcag taccagaacg acctgaaggg cctggtgttc 1860

gacaaccagg acgacgagct ccccaagatc cccggcgagt acatcctgga gaacgtgttc 1920

cagatcgacg tgagccgatc caagtggctg gacctcgccg tgctcttcag catgatcttc 1980

atctaccgcc tcctcttctt cgccatgatc aaggtcagcg aggacgtcac cccgtgggtg 2040

cgcggctacg tcgcgcggcg ccgggtgcag ggcaagggcg cccgcggccg cggggcggac 2100

ctctccgccg cgcgctcgcc gtcgctccgc gcctacgtcg tcgacgccgc cgacgacctg 2160

ccgccggcct ga 2172

<210> 6

<211> 723

<212> PRT

<213> Rice (Oryza sativa Japonica)

<400> 6

Met Lys Lys Gln Gly Gly Gly Gly Gly Val Ala Ala Ala Ala Val Ala

1 5 10 15

Ala Ala Gly Met Val Lys Ala Glu Leu Glu Asp Val Gly Ile Lys Ala

20 25 30

Ala Gly Gly Ala Val Ala Ala Leu Ser Pro Leu Ser Glu Thr Leu Trp

35 40 45

Arg Glu Lys Ala Ala Ala Glu Phe Leu Gly Asp Val Ser Ala Arg Leu

50 55 60

Ala Trp Arg Asp Leu Thr Val Thr Val Val Leu Gly Gly Gly Gly Gly

65 70 75 80

Gly Gly Gly Gly Gly Thr Thr Gln Asp Val Leu Gln Gly Leu Thr Gly

85 90 95

His Ala Glu Pro Gly Thr Ile Thr Ala Leu Met Gly Pro Ser Gly Ser

100 105 110

Gly Lys Ser Thr Leu Leu Asp Ala Leu Ala Gly Arg Leu Ala Ala Asn

115 120 125

Ala Phe Leu Asp Gly Thr Val Leu Leu Asn Gly Arg Lys Ala Asn Leu

130 135 140

Ser Phe Gly Ala Ala Ala Tyr Val Thr Gln Asp Asp Asn Leu Ile Gly

145 150 155 160

Thr Leu Thr Val Arg Glu Thr Ile Ser Tyr Ser Ala Arg Leu Arg Leu

165 170 175

Pro Asp Lys Met Pro Met Glu Glu Lys Arg Ala Leu Val Glu Gly Thr

180 185 190

Ile Val Glu Met Gly Leu Gln Asp Cys Ala Asp Thr Val Val Gly Asn

195 200 205

Trp His Leu Arg Gly Ile Ser Gly Gly Glu Lys Arg Arg Val Ser Ile

210 215 220

Ala Leu Glu Ile Leu Met Arg Pro Arg Leu Leu Phe Leu Asp Glu Pro

225 230 235 240

Thr Ser Gly Leu Asp Ser Ala Ser Ala Phe Phe Val Thr Gln Thr Leu

245 250 255

Arg Gly Leu Ala Arg Asp Gly Arg Thr Val Ile Ala Ser Ile His Gln

260 265 270

Pro Ser Ser Glu Val Phe Glu Leu Phe Asp Arg Leu Tyr Leu Leu Ser

275 280 285

Gly Gly Lys Thr Val Tyr Phe Gly Leu Ala Ser Glu Ala Cys Gln Phe

290 295 300

Phe Ala Gln Ala Gly Phe Pro Cys Pro Pro Leu Arg Asn Pro Ser Asp

305 310 315 320

His Phe Leu Arg Cys Ile Asn Ala Asp Phe Asp Lys Val Lys Ala Thr

325 330 335

Leu Lys Gly Ser Met Lys Arg Arg Phe Glu Arg Ser Asp Asp Pro Leu

340 345 350

Asp Arg Ile Met Thr Ser Glu Ala Ile Arg Arg Leu Ile Thr Tyr Tyr

355 360 365

Lys Asn Ser Gln Tyr Tyr Phe Ala Ala Gln Gln Lys Val Asn Glu Met

370 375 380

Ala Arg Val Lys Gly Thr Val Leu Asp Ala Gly Gly Ser Gln Ala Ser

385 390 395 400

Phe Trp Met Gln Ala Phe Thr Leu Thr Lys Arg Ser Phe Ile Asn Met

405 410 415

Ser Arg Asp Phe Gly Tyr Tyr Trp Leu Arg Leu Ile Ile Tyr Ile Val

420 425 430

Val Thr Val Cys Ile Gly Thr Ile Tyr Leu Asn Val Gly Thr Arg Tyr

435 440 445

Ser Ser Ile Leu Ala Arg Gly Ala Cys Ala Ser Phe Val Phe Gly Phe

450 455 460

Val Thr Phe Met Ser Ile Gly Gly Phe Pro Ser Phe Val Glu Asp Met

465 470 475 480

Lys Val Phe Gln Arg Glu Arg Leu Asn Gly His Tyr Gly Val Leu Ala

485 490 495

Phe Val Ile Ser Asn Thr Ile Ser Ala Met Pro Phe Leu Ile Leu Ile

500 505 510

Thr Phe Ile Ser Gly Thr Met Cys Tyr Phe Met Val Arg Leu His Pro

515 520 525

Gly Phe Thr His Tyr Leu Phe Phe Val Leu Cys Leu Tyr Ala Ser Val

530 535 540

Thr Val Val Glu Ser Leu Met Met Ala Ile Ala Ser Val Ile Pro Asn

545 550 555 560

Phe Leu Met Gly Ile Ile Ile Gly Ala Gly Ile Gln Gly Ile Phe Met

565 570 575

Leu Val Ser Gly Tyr Phe Arg Leu Pro His Asp Ile Pro Lys Pro Phe

580 585 590

Trp Arg Tyr Pro Met Ser Tyr Ile Ser Phe His Tyr Trp Ala Leu Gln

595 600 605

Gly Gln Tyr Gln Asn Asp Leu Lys Gly Leu Val Phe Asp Asn Gln Asp

610 615 620

Asp Glu Leu Pro Lys Ile Pro Gly Glu Tyr Ile Leu Glu Asn Val Phe

625 630 635 640

Gln Ile Asp Val Ser Arg Ser Lys Trp Leu Asp Leu Ala Val Leu Phe

645 650 655

Ser Met Ile Phe Ile Tyr Arg Leu Leu Phe Phe Ala Met Ile Lys Val

660 665 670

Ser Glu Asp Val Thr Pro Trp Val Arg Gly Tyr Val Ala Arg Arg Arg

675 680 685

Val Gln Gly Lys Gly Ala Arg Gly Arg Gly Ala Asp Leu Ser Ala Ala

690 695 700

Arg Ser Pro Ser Leu Arg Ala Tyr Val Val Asp Ala Ala Asp Asp Leu

705 710 715 720

Pro Pro Ala

<210> 7

<211> 2392

<212>DNA

<213> Rice (Oryza sativa)

<400> 7

atgaagaagc agggaggagg aggaggagta gcggcggcgg cggtggcggc ggcggggatg 60

gtgaaggcgg agctggagga cgtggggatc aaggcggcgg gaggggcggt ggcggcgctg 120

agcccgctga gcgagacgct gtggcgggag aaggcggcgg cggagttcct cggcgacgtg 180

tcggcgcgcc tggcgtggcg cgacctcacc gtcaccgtcg tgctcggcgg cggcggcggc 240

ggcggcggag gagggacgac gcaggacgtg ctccaggggc tcacggggca cgccgagccg 300

ggcaccatca cggcgctcat gggcccctcc ggctccggca agtccaccct gctcgacgcc 360

ctcgccggcc gcctcgccgc caacgccttc ctcgacggca ccgtcctcct caacggccgc 420

aaggccaacc tctccttcgg cgccgcggcc tatgtgaccc aagatgacaa cttgatcggc 480

acattgacgg tgagggagac catctcatac tcggcgcggc ttcgccttcc cgacaagatg 540

cccatggaag agaagcgcgc ccttgtcgag ggcacaatcg tcgagatggg gctgcaggac 600

tgcgccgaca cggtggtcgg caattggcac ctgaggggaa tcagtggtgg tgagaagagg 660

agggtcagca ttgcccttga gatactgatg aggcccaggc tgctcttcct ggatgaacca 720

accagtggcc ttgacagtgc ttcagcgttc ttcgtgacac agacactgcg tgggctagcg 780

agggatggca ggacagtgat agcatcgatc catcagccga gcagcgaggt ttttgagctt 840

ttcgaccgct tgtatctcct ttcaggaggc aaaactgttt actttgggct ggcctctgaa 900

gcttgtcagt tctttgccca agccggcttc ccttgcccac cgctgcgcaa tccatcggat 960

catttcctca gatgcataaa cgcagacttc gacaaggtga aggccactct gaaaggatca 1020

atgaagagaa gatttgaaag atccgatgac ccacttgatc ggattatgac ttcggaggcc 1080

atcagaaggc tgatcaccta ctacaagaac tcacagtact actttgctgc gcagcagaaa 1140

gtcaatgaga tggcacgggt taaagggaca gtgctggatg caggagggag ccaggctagc 1200

ttttggatgc aggctttcac actcacgaag cgatcgttca tcaacatgtc gcgggacttc 1260

gggtactact ggctgaggct tatcatctac attgttgtga cagtctgcat cgggacaatc 1320

tacctcaatg ttggcaccag atacagctca attctggcaa gaggtgcatg cgcttctttc 1380

gtcttcggct ttgtcacgtt catgtcgatt ggaggattcc catcttttgt ggaagacatg 1440

aaggtgtttc agagagagag gctcaatggg cactatggcg tgctggcatt cgtgataagc 1500

aacacgatct cggcgatgcc attcttgatc ttgatcacct tcatctccgg gacaatgtgc 1560

tacttcatgg tgcgcctcca cccgggcttc acgcactacc tcttcttcgt cctctgcctc 1620

tacgcaagcg tcaccgtcgt cgagagcttg atgatggcca ttgccagtgt catccccaac 1680

ttcctcatgg gcatcatcat cggtgcagga attcagatga attaattacc atccatcaca 1740

acaacttgat cctgtccatc tttatcaggc actgtttgga gtcctgcaac gcatacaaac 1800

tgggattaaa ttttgacact gtacttggaa ctgcacaaca taatgttgat gtttgttaat 1860

attctctcat gatcatgaca tgcaacaaca tttattgaag agaaaaatat cttttggaaa 1920

tggacagggg atattcatgc tggtttcagg atacttcagg cttccccatg acatcccaaa 1980

gccattctgg aggtatccca tgtcatacat cagcttccac tactgggcat tgcagggtca 2040

gtaccagaac gacctgaagg gcctggtgtt cgacaaccag gacgacgagc tccccaagat 2100

ccccggcgag tacatcctgg agaacgtgtt ccagatcgac gtgagccgat ccaagtggct 2160

ggacctcgcc gtgctcttca gcatgatctt catctaccgc ctcctcttct tcgccatgat 2220

caaggtcagc gaggacgtca ccccgtgggt gcgcggctac gtcgcgcggc gccgggtgca 2280

gggcaagggc gcccgcggcg cccgcggccg cggggcggac ctcgccgccg cgcgctcgcc 2340

gtcgctccgc gcctacgtcg tcgacgccgc cgacgacctg ccgccggcct ga 2392

<210> 8

<211> 574

<212> PRT

<213> Rice (Oryza sativa)

<400> 8

Met Lys Lys Gln Gly Gly Gly Gly Gly Val Ala Ala Ala Ala Val Ala

1 5 10 15

Ala Ala Gly Met Val Lys Ala Glu Leu Glu Asp Val Gly Ile Lys Ala

20 25 30

Ala Gly Gly Ala Val Ala Ala Leu Ser Pro Leu Ser Glu Thr Leu Trp

35 40 45

Arg Glu Lys Ala Ala Ala Glu Phe Leu Gly Asp Val Ser Ala Arg Leu

50 55 60

Ala Trp Arg Asp Leu Thr Val Thr Val Val Leu Gly Gly Gly Gly Gly

65 70 75 80

Gly Gly Gly Gly Gly Thr Thr Gln Asp Val Leu Gln Gly Leu Thr Gly

85 90 95

His Ala Glu Pro Gly Thr Ile Thr Ala Leu Met Gly Pro Ser Gly Ser

100 105 110

Gly Lys Ser Thr Leu Leu Asp Ala Leu Ala Gly Arg Leu Ala Ala Asn

115 120 125

Ala Phe Leu Asp Gly Thr Val Leu Leu Asn Gly Arg Lys Ala Asn Leu

130 135 140

Ser Phe Gly Ala Ala Ala Tyr Val Thr Gln Asp Asp Asn Leu Ile Gly

145 150 155 160

Thr Leu Thr Val Arg Glu Thr Ile Ser Tyr Ser Ala Arg Leu Arg Leu

165 170 175

Pro Asp Lys Met Pro Met Glu Glu Lys Arg Ala Leu Val Glu Gly Thr

180 185 190

Ile Val Glu Met Gly Leu Gln Asp Cys Ala Asp Thr Val Val Gly Asn

195 200 205

Trp His Leu Arg Gly Ile Ser Gly Gly Glu Lys Arg Arg Val Ser Ile

210 215 220

Ala Leu Glu Ile Leu Met Arg Pro Arg Leu Leu Phe Leu Asp Glu Pro

225 230 235 240

Thr Ser Gly Leu Asp Ser Ala Ser Ala Phe Phe Val Thr Gln Thr Leu

245 250 255

Arg Gly Leu Ala Arg Asp Gly Arg Thr Val Ile Ala Ser Ile His Gln

260 265 270

Pro Ser Ser Glu Val Phe Glu Leu Phe Asp Arg Leu Tyr Leu Leu Ser

275 280 285

Gly Gly Lys Thr Val Tyr Phe Gly Leu Ala Ser Glu Ala Cys Gln Phe

290 295 300

Phe Ala Gln Ala Gly Phe Pro Cys Pro Pro Leu Arg Asn Pro Ser Asp

305 310 315 320

His Phe Leu Arg Cys Ile Asn Ala Asp Phe Asp Lys Val Lys Ala Thr

325 330 335

Leu Lys Gly Ser Met Lys Arg Arg Phe Glu Arg Ser Asp Asp Pro Leu

340 345 350

Asp Arg Ile Met Thr Ser Glu Ala Ile Arg Arg Leu Ile Thr Tyr Tyr

355 360 365

Lys Asn Ser Gln Tyr Tyr Phe Ala Ala Gln Gln Lys Val Asn Glu Met

370 375 380

Ala Arg Val Lys Gly Thr Val Leu Asp Ala Gly Gly Ser Gln Ala Ser

385 390 395 400

Phe Trp Met Gln Ala Phe Thr Leu Thr Lys Arg Ser Phe Ile Asn Met

405 410 415

Ser Arg Asp Phe Gly Tyr Tyr Trp Leu Arg Leu Ile Ile Tyr Ile Val

420 425 430

Val Thr Val Cys Ile Gly Thr Ile Tyr Leu Asn Val Gly Thr Arg Tyr

435 440 445

Ser Ser Ile Leu Ala Arg Gly Ala Cys Ala Ser Phe Val Phe Gly Phe

450 455 460

Val Thr Phe Met Ser Ile Gly Gly Phe Pro Ser Phe Val Glu Asp Met

465 470 475 480

Lys Val Phe Gln Arg Glu Arg Leu Asn Gly His Tyr Gly Val Leu Ala

485 490 495

Phe Val Ile Ser Asn Thr Ile Ser Ala Met Pro Phe Leu Ile Leu Ile

500 505 510

Thr Phe Ile Ser Gly Thr Met Cys Tyr Phe Met Val Arg Leu His Pro

515 520 525

Gly Phe Thr His Tyr Leu Phe Phe Val Leu Cys Leu Tyr Ala Ser Val

530 535 540

Thr Val Val Glu Ser Leu Met Met Ala Ile Ala Ser Val Ile Pro Asn

545 550 555 560

Phe Leu Met Gly Ile Ile Ile Gly Ala Gly Ile Gln Met Asn

565 570

<210> 9

<211> 950

<212>DNA

<213> Rice (Oryza sativa)

<400> 9

accatatggc tgttcacaac ccaacaactt gcaatccagt gcatgcatta catggccaag 60

aatccagcgg ttgagccact gacatgtata ttccactttc gttatgaccc gcatgtcagt 120

ggctcaattg cacatacagc taagtcagag aatttccttc atctctacca atctattctc 180

tctctttggc ctggtttagt cccaaaacat catatcgaat ctttgaagac atgcatgtag 240

cattaaatat ataataaaaa aaactaattg catagttagg cggaaatcac gagacgaatc 300

ttttgagcct aataagtcca ttattagccg taagtgctac agtaacccac atacgctaat 360

gacggattaa ttaagctcaa aagattcgtc ttgcgatttt caggcaagtt atgaaattag 420

ttttttcatt cgtgtccgaa aacccctacc gatatccagt caaacatccg atgtgacacc 480

caaacatttc ttttcttgaa ttaaacatgc cctttctctt gtgcttgcat tgcatatgca 540

tatgcacaat gcattgcttc tagatcacac caaacagcta ctagcccccc cttatgacat 600

gccattcatc tccattgcat gcttgcatat tgcatgcatg ctgctgctgc ttctactcct 660

ccaactctct agagctagct accttgaccc taggtcaaac ccctacaccc ctcattactt 720

ctgcacctga tctccttctc tctctagctt ctctccctag attttctctc tctaacctgc 780

tcctccatta ctggccttta tatccaccac ctcctcctct tctcctccct cataggtggt 840

gtttgcttag ctagctagct agctccatca ctagctctcg catcgcatgc cacggtgagc 900

cggtgtcgcc ggcgtcgtac ggacgtagca agcgagagag ggagagatga 950

<210> 10

<211> 2631

<212>DNA

<213> Barley (Hordeum vulgare)

<400> 10

agagagagag agagcgcaca gagcagccta tagttgcagc ttggacacgc tgcccaagaa 60

acgccattct tctcccgcgc cggcctggct tgtacggtca gctagcttcc tccccacacgc 120

agccgcggcg aggaggaagg cgccgtgtgc gagcgttccc accaccttgt taccaagaag 180

aagccccggc ttgccctcgt ctgccctgct ctgtcacggc ctcccaaggt taggtcgtgg 240

tgtctggccg gcgaggctat gaagaaggga ggggcgggga cggcgaagga tgcggcggcg 300

gtggggcagg tgaaggccga gctggaggac gtggggaagg cggcggggcg ggcggtggcg 360

gtggcggcgg cgccggcgct gagcccgctg agcgagacgc tgtggcggga gaagacggcg 420

gtggaggttcc tcggggacgt gtcagcgcgg ctggcctgga gggacctcac ggtcaccgtc 480

gcgctcggca gcggcgacac gcagaccgtg ctcgaggggc tcacggggta cgccgagccg 540

ggcaccatca ccgcgctcat gggcccctcc ggctccggca agtccacgct gctcgacgcc 600

ctcgccggcc gcctcgccgc caacgccttc ctctccggga ccatcctgct caacggacgc 660

aaggccaatc tttccttcgg cgccgcggcc tatgtgacgc aagacgacaa cttgattggc 720

actttgacgg tgagggaaac gatctcgtac tcggcacgcc ttcgactccc cgacaacatg 780

cccatggaag agaagcgtgc cttggtggag ggcaccatcg tggagatggg gcttcaggac 840

tgcgcagaca cggtcgtcgg caattggcac ctgagggggg tcagcggcgg cgagaagagg 900

agggtcagca ttgccctgga gatactgatg aggcctaggc tgctcttcct ggatgagccc 960

accagtggcc tcgacagtgc ttcggcgttc ttcgtgacac aaactctgcg tgggctggcc 1020

agagatggcc ggactgtgat cgcgtcgatc caccagccca gcagcgaggt cttcgagctt 1080

ttcgatcgcc tctatcttct ctcagggggc aaaacagtct actttgggca ggcttccgag 1140

gcttgcgagt tctttgccca agctggcttc ccgtgcccgc cgctgcgcaa cccgtcggat 1200

catttcctcc ggtgcataaa cgcggacttt gacaaggtga aggccaacact gaaaggatca 1260

atgaagataa ggtttgagag gtccgacgat cccctcgagc acaccactac ttcggacgcg 1320

atcagaaggc tgttcagcta ctaccagcac tcgcagcact acttgacggc actgcagaag 1380

gtcgacgaga tggcacgggt caaaggaacg gtcctggacg caggggggag ccaggccagc 1440

ttcgggatgc aggccttcac gcttaccaag cgatcgttcg tcaacatgtc gagggacttt 1500

ggatactact ggctgaggct tgtcatctac attttggtca cggtctgcat tgggtccatc 1560

tacctcaacg tcggcaccaa atacagctcc atcctggcaa ggggcgcgtg cgcgtccttc 1620

atcttcggct tcgtcacgtt catgtcgatc ggagggttcc cgtctttcgt ggaggacatg 1680

aaggtgttcc aggggagcg gctgaacggg cactacggcg tgctggcgtt cgtgatgagc 1740

aacacgctgt cagcgatgcc gttcctgatc ctcatcacgt tcctgtcggg gacgctgtgc 1800

tacttcatgg tgcgcctcca cccgggcttc acgcactacc tcttcttcct cctctgcctt 1860

tacgccagcg tcaccgtcgt cgagagcctt atgatggcca tcgccagcat catccccaac 1920

ttcctaatgg gcatcatcat cggcgcaggg atacagggga tatcatgct ggtgtcgggg 1980

tacttcaggc tcccacatga catcccgaag cccttctgga ggtaccccat gtcctacatc 2040

agcttccact actgggcact gcaggggcag taccagaacg acctggtggg gctggtgttc 2100

gacaaccagg acgacgagct gcccaagatc ccgggggagt acatcctgga gaacgtgttc 2160

cagatcgacg tgagccggtc caagtggctg gacctgtcgg tgctcttcgg catgatcgtc 2220

atctaccgcc tgctcttctt tgccatgatc aaggtcagcg aggacgtgac gccgtgggtg 2280

cgcggctacg tcgccaggag gagggtgcag cacaagcgca gagaggcgga gctcgccatg 2340

gtccggacgc cgtcgctgcg cggctacgtc atcgacgcgg cgccggagct gccggccgat 2400

catccgtgat ccatcatcga gaatggatga tgaatggcct gagctgaatc ggtatacatg 2460

agccgtgatg tgtgatcctt gtgtggtgtg actaccatct gacgatgcat cattgaaatg 2520

gcatcttgta gcagtaaggt tgtcactcaa atagtgctcc gaatggctac aacttttatc 2580

aagtgtagta ctacaatctg atggttcatc attaaggtgg catcttgtag c 2631

<210> 11

<211> 2151

<212>DNA

<213> Barley (Hordeum vulgare)

<400> 11

atgaagaagg gaggggcggg gacggcgaag gatgcggcgg cggtggggca ggtgaaggcc 60

gagctggagg acgtggggaa ggcggcgggg cgggcggtgg cggtggcggc ggcgccggcg 120

ctgagcccgc tgagcgagac gctgtggcgg gagaagacgg cggtggagtt cctcggggac 180

gtgtcagcgc ggctggcctg gagggacctc acggtcaccg tcgcgctcgg cagcggcgac 240

acgcagaccg tgctcgaggg gctcacgggg tacgccgagc cgggcaccat caccgcgctc 300

atgggcccct ccggctccgg caagtccacg ctgctcgacg ccctcgccgg ccgcctcgcc 360

gccaacgcct tcctctccgg gaccatcctg ctcaacggac gcaaggccaa tctttccttc 420

ggcgccgcgg cctatgtgac gcaagacgac aacttgattg gcactttgac ggtgagggaa 480

acgatctcgt actcggcacg ccttcgactc cccgacaaca tgcccatgga agagaagcgt 540

gccttggtgg agggcaccat cgtggagatg gggcttcagg actgcgcaga cacggtcgtc 600

ggcaattggc acctgagggg ggtcagcggc ggcgagaaga ggagggtcag cattgccctg 660

gagatactga tgaggcctag gctgctcttc ctggatgagc ccaccagtgg cctcgacagt 720

gcttcggcgt tcttcgtgac acaaactctg cgtggggctgg ccagagatgg ccggactgtg 780

atcgcgtcga tccaccagcc cagcagcgag gtcttcgagc ttttcgatcg cctctatctt 840

ctctcagggg gcaaaacagt ctactttgggg caggcttccg aggcttgcga gttctttgcc 900

caagctggct tcccgtgccc gccgctgcgc aacccgtcgg atcatttcct ccggtgcata 960

aacgcggact ttgacaaggt gaaggccaca ctgaaaggat caatgaagat aaggtttgag 1020

aggtccgacg atcccctcga gcacaccact acttcggacg cgatcagaag gctgttcagc 1080

tactaccagc actcgcagca ctacttgacg gcactgcaga aggtcgacga gatggcacgg 1140

gtcaaaggaa cggtcctgga cgcagggggg agccaggcca gcttcgggat gcaggccttc 1200

acgcttacca agcgatcgtt cgtcaacatg tcgagggact ttggatacta ctggctgagg 1260

cttgtcatct aattttggt cacggtctgc attgggtcca tctacctcaa cgtcggcacc 1320

aaatacagct ccatcctggc aaggggcgcg tgcgcgtcct tcatcttcgg cttcgtcacg 1380

ttcatgtcga tcggagggtt cccgtctttc gtggaggaca tgaaggtgtt ccagaggggag 1440

cggctgaacg ggcactacgg cgtgctggcg ttcgtgatga gcaacacgct gtcagcgatg 1500

ccgttcctga tcctcatcac gttcctgtcg gggacgctgt gctacttcat ggtgcgcctc 1560

cacccgggct tcacgcacta cctcttcttc ctcctctgcc tttacgccag cgtcaccgtc 1620

gtcgagagcc ttatgatggc catcgccagc atcatcccca acttcctaat gggcatcatc 1680

atcggcgcag ggatacagggg gatattcatg ctggtgtcgg ggtacttcag gctcccacat 1740

gacatcccga agcccttctg gaggtacccc atgtcctaca tcagcttcca ctactgggca 1800

ctgcaggggc agtaccagaa cgacctggtg gggctggtgt tcgacaacca ggacgacgag 1860

ctgcccaaga tcccggggga gtacatcctg gagaacgtgt tccagatcga cgtgagccgg 1920

tccaagtggc tggacctgtc ggtgctcttc ggcatgatcg tcatctaccg cctgctcttc 1980

tttgccatga tcaaggtcag cgaggacgtg acgccgtggg tgcgcggcta cgtcgccagg 2040

aggagggtgc agcacaagcg cagagaggcg gagctcgcca tggtccggac gccgtcgctg 2100

cgcggctacg tcatcgacgc ggcgccggag ctgccggccg atcatccgtg a 2151

<210> 12

<211> 716

<212> PRT

<213> Barley (Hordeum vulgare)

<400> 12

Met Lys Lys Gly Gly Ala Gly Thr Ala Lys Asp Ala Ala Ala Val Gly

1 5 10 15

Gln Val Lys Ala Glu Leu Glu Asp Val Gly Lys Ala Ala Gly Arg Ala

20 25 30

Val Ala Val Ala Ala Ala Pro Ala Leu Ser Pro Leu Ser Glu Thr Leu

35 40 45

Trp Arg Glu Lys Thr Ala Val Glu Phe Leu Gly Asp Val Ser Ala Arg

50 55 60

Leu Ala Trp Arg Asp Leu Thr Val Thr Val Ala Leu Gly Ser Gly Asp

65 70 75 80

Thr Gln Thr Val Leu Glu Gly Leu Thr Gly Tyr Ala Glu Pro Gly Thr

85 90 95

Ile Thr Ala Leu Met Gly Pro Ser Gly Ser Gly Lys Ser Thr Leu Leu

100 105 110

Asp Ala Leu Ala Gly Arg Leu Ala Ala Asn Ala Phe Leu Ser Gly Thr

115 120 125

Ile Leu Leu Asn Gly Arg Lys Ala Asn Leu Ser Phe Gly Ala Ala Ala

130 135 140

Tyr Val Thr Gln Asp Asp Asn Leu Ile Gly Thr Leu Thr Val Arg Glu

145 150 155 160

Thr Ile Ser Tyr Ser Ala Arg Leu Arg Leu Pro Asp Asn Met Pro Met

165 170 175

Glu Glu Lys Arg Ala Leu Val Glu Gly Thr Ile Val Glu Met Gly Leu

180 185 190

Gln Asp Cys Ala Asp Thr Val Val Gly Asn Trp His Leu Arg Gly Val

195 200 205

Ser Gly Gly Glu Lys Arg Arg Val Ser Ile Ala Leu Glu Ile Leu Met

210 215 220

Arg Pro Arg Leu Leu Phe Leu Asp Glu Pro Thr Ser Gly Leu Asp Ser

225 230 235 240

Ala Ser Ala Phe Phe Val Thr Gln Thr Leu Arg Gly Leu Ala Arg Asp

245 250 255

Gly Arg Thr Val Ile Ala Ser Ile His Gln Pro Ser Ser Glu Val Phe

260 265 270

Glu Leu Phe Asp Arg Leu Tyr Leu Leu Ser Gly Gly Lys Thr Val Tyr

275 280 285

Phe Gly Gln Ala Ser Glu Ala Cys Glu Phe Phe Ala Gln Ala Gly Phe

290 295 300

Pro Cys Pro Pro Leu Arg Asn Pro Ser Asp His Phe Leu Arg Cys Ile

305 310 315 320

Asn Ala Asp Phe Asp Lys Val Lys Ala Thr Leu Lys Gly Ser Met Lys

325 330 335

Ile Arg Phe Glu Arg Ser Asp Asp Pro Leu Glu His Thr Thr Thr Ser

340 345 350

Asp Ala Ile Arg Arg Leu Phe Ser Tyr Tyr Gln His Ser Gln His Tyr

355 360 365

Leu Thr Ala Leu Gln Lys Val Asp Glu Met Ala Arg Val Lys Gly Thr

370 375 380

Val Leu Asp Ala Gly Gly Ser Gln Ala Ser Phe Gly Met Gln Ala Phe

385 390 395 400

Thr Leu Thr Lys Arg Ser Phe Val Asn Met Ser Arg Asp Phe Gly Tyr

405 410 415

Tyr Trp Leu Arg Leu Val Ile Tyr Ile Leu Val Thr Val Cys Ile Gly

420 425 430

Ser Ile Tyr Leu Asn Val Gly Thr Lys Tyr Ser Ser Ile Leu Ala Arg

435 440 445

Gly Ala Cys Ala Ser Phe Ile Phe Gly Phe Val Thr Phe Met Ser Ile

450 455 460

Gly Gly Phe Pro Ser Phe Val Glu Asp Met Lys Val Phe Gln Arg Glu

465 470 475 480

Arg Leu Asn Gly His Tyr Gly Val Leu Ala Phe Val Met Ser Asn Thr

485 490 495

Leu Ser Ala Met Pro Phe Leu Ile Leu Ile Thr Phe Leu Ser Gly Thr

500 505 510

Leu Cys Tyr Phe Met Val Arg Leu His Pro Gly Phe Thr His Tyr Leu

515 520 525

Phe Phe Leu Leu Cys Leu Tyr Ala Ser Val Thr Val Val Glu Ser Leu

530 535 540

Met Met Ala Ile Ala Ser Ile Ile Pro Asn Phe Leu Met Gly Ile Ile

545 550 555 560

Ile Gly Ala Gly Ile Gln Gly Ile Phe Met Leu Val Ser Gly Tyr Phe

565 570 575

Arg Leu Pro His Asp Ile Pro Lys Pro Phe Trp Arg Tyr Pro Met Ser

580 585 590

Tyr Ile Ser Phe His Tyr Trp Ala Leu Gln Gly Gln Tyr Gln Asn Asp

595 600 605

Leu Val Gly Leu Val Phe Asp Asn Gln Asp Asp Glu Leu Pro Lys Ile

610 615 620

Pro Gly Glu Tyr Ile Leu Glu Asn Val Phe Gln Ile Asp Val Ser Arg

625 630 635 640

Ser Lys Trp Leu Asp Leu Ser Val Leu Phe Gly Met Ile Val Ile Tyr

645 650 655

Arg Leu Leu Phe Phe Ala Met Ile Lys Val Ser Glu Asp Val Thr Pro

660 665 670

Trp Val Arg Gly Tyr Val Ala Arg Arg Arg Val Gln His Lys Arg Arg

675 680 685

Glu Ala Glu Leu Ala Met Val Arg Thr Pro Ser Leu Arg Gly Tyr Val

690 695 700

Ile Asp Ala Ala Pro Glu Leu Pro Ala Asp His Pro

705 710 715

<210> 13

<211> 5321

<212>DNA

<213> Sorghum (Sorghum bicolor)

<400> 13

accgcatcga ccggccggtt cctctctccc gacacgacgc gccccagccc agtgcacggc 60

cccgccacgc tagcctccca gcagctccca gccccagctc cgctcagggc gcgcctgcag 120

ttccggccat gagacggggcc ggaggaggag acctgcaggg cgaccacggc acgacgcctc 180

ggtccgcggc ggcagggcag gccatggtgg agctgcaggc caacgggtcg gcggccgccg 240

ccgggggcgc catggtggtg ggtctcagcc cgctcagcga gaccctgtgg cgggacagca 300

aggcgctccc gccgggtgcc ggccccgccg cgctcatcgg cgacgtgtcc gccaggctca 360

cgtggaagga cctctgcgtc accgtggcgc tcggccccgg caagacgcag accgtgctgg 420

acgaactcac cgggtacgcg gagcccggct cgctcaccgc gctcatgggc ccctccgggt 480

ccggcaagtc caccctgctc gacgccctcg ccggccgcct cgccgccaac gccttcctct 540

ccggcgacgt gctcctcaac ggccgcaagg ccaagctctc cttcggcgcc gcggtacgtg 600

acctttgctg cttcctcagt cctcgcttaa gctcctagct atagatcatg cagcactact 660

ctctctcaac tgggaaaaaa tagaatcact gttctatttt tgagcaccaa aaacactccg 720

tactgctact acttcagcca gatcatccat atataggagc tagcctagca gttccgttaa 780

gggtttctta ggtagcatct gttcaagttc gaaactatct tccatatata actggcatct 840

cgcaacatgc aaatcatatt tgaaggtatc cggatgaccg gatctggtaa tctacttggt 900

gtgtcagtaa aacaatcgtt gctggtaatt aaagtgtggt ttgtctatgg tttttccaaa 960

tgaaatatac tatatatatg ctgcgtcctg ttattatcac tataatcaat gacataacta 1020

gtactctcat tctgttcttt tttaaaagac ctaccacaac acagctacgc tttgaccact 1080

atagtagtgc tataattaag cagaggtgcg agctgcttat ttcatgtcgg gcagtaagca 1140

gggagatcgg ctagccggca tagtgttatt agcgacagat cacagataca cgtccaaatc 1200

caatcgattc gggtcgattt ttggcgtccg gccaagatgt cgaaagcacg tacggctagg 1260

ctttgcatag tagagctccg gcgtgtaatg ttgttaggtg cgcggattta attcgccctc 1320

gatcggttgg tcgatggcgc ctccactgga attcattcaa acacataggt cgtactgcgt 1380

actgctgcta gtatgattag tatcgtgatt ttgatctctc acatcgatag atccagaggc 1440

gaaaaacatg gcgatcgata tcgtcctgct ctgctgtcaa tacaatacac atggcgatcg 1500

atatccacaa cggctgaagc cacctacaca gcataaatac tgacagctgg gtttttctct 1560

ctgatcctgc tgcgaattta tggtgatacg aaccgtccgg gctgtggcgt ttgggtacgt 1620

ctcaccagca gttacacgca acgggatcct gcaggcgcat caaacgcatc agttttctgg 1680

ctaatcttgc tctgatccag agtagaaagt tttaagtgga cgagtttttt tttttatggg 1740

agagtttgtg ctactactgc cggaggagga gactgatcca gaggcgttgt tggtgccgtg 1800

tgtatacgcg cgcgtacgtg cttcttctgc ttgcaggcgt acgtgacgca ggacgacaac 1860

ctgatcggga cgctgacggt gcgcgagacg atcggctact cggcgctgct gcggctgccg 1920

gacaagatgc cgcgggagga caagcgcgcg ctggtggagg gcaccatcgt cgagatgggg 1980

ctccaggact gcgccgacac cgtcatcggg aactggcacc tccgcggggt cagcggcggc 2040

gagaagcgcc gcgtcagcat cgcgctcgag ctcctcatgc gcccgcgcct cctcttcctc 2100

gacgagccca ccagcggcct cgacaggtga gcagcacggt ggcggccgtg gtgcacggcg 2160

gcaaattgtg tgagcctgca atgtcagttg gtggtggtga ggctaactta tttagctgcg 2220

ggcgtggctg tatgtgtgca gctcctcggc gttcttcgtg acgcagacgc tgcgaggcct 2280

ggccagggac ggcaggacag tgattgcttc catccaccag cccagcagcg aggtgttcga 2340

gctcttcgac atgctcttcc tgctgtccgg gggcaaaacc gtctacttcg gccaagcatc 2400

gcaagcatgt gaggtagtaa tcctgtattc ttgccggtgt taccgtgaga tgccgaatgc 2460

atgcttcggc gacaatgtga tcgtgcgact gactctcctt gcattgctcg gttgcagttc 2520

tttgctcaag gcggtttccc ttgcccgcct ctgaggaacc catcggacca tttcctgagg 2580

tgcgtcaact cggacttcga caaggtgaag gccaacactga aaggatcaat gaaggcaagg 2640

gtaagaagaa gaaggtccag gggcaaatca gaggccacca aaacatgaaa aaaaaagcct 2700

tgttctctgc tgtgaataaa atactaatac gaccttttat tttgtgttca tgaaaaggct 2760

gagaggtccg acgatccact cgataggatg actacatcag aagccatcag gaaattggtt 2820

ggatcctaca gcaggtccca atactactac gctgcgaggg agaaagttaa tgatatctcg 2880

agaatggtac gtattctgag tcagcaaaca caacattatg cattgatatg ttgtatctgt 2940

ttcctgtgtc cacataataa ataccgcatc tggggctggg aaaaactaac cataaccatc 3000

ttgttcctta cagaaaggaa ctgtgctgga ttcaggtggc agccaggcta gcttcctgat 3060

gcaggcttgc accctgacca agcgctcgtt catcaacatg tcacgggact ttgggtatta 3120

ctggctcagg ctcctgatct atctcctcgt gactgtttgc atcggcacaa tctacctgga 3180

tgttggcact aaatacacat ccatcctggt aagtaggaag gaaagaaatt ggtccggaag 3240

ttctgggctg gactgatatg aatgcacaat aatgtagtca catgaccaat gtcatgaagc 3300

agcatatggt tgattttaat gcattcactc taccttgcct ctgcaggcca gagcttcctg 3360

ttctgcattc gtctttgggt tcgttacgtt catgtccatt ggaggattcc cttcgtttgt 3420

tgaagaaatg aaggtgaaat taaaatggtg ttttttcatc tgtaaattag aaatgctatg 3480

aattttgctc acctttcgtt tgcttcattt atacaggtct tccaacggga acggctgaat 3540

gggcactatg gtgtcgcggc atttgtcatc agtaacacca tctcagcaac gccattcctg 3600

attctaatat gcttcttgtc gggaactata tgctacttta tggtgcgcct gcatcctggt 3660

ttcgaacact atatcttctt cgtcttgaac ctgtatgcca gcgtcacagt ggtcgaaagc 3720

ttaatgatgg caattgccag tgtcattccc aatttcctca tgggtatcat cataggggct 3780

ggaattcagg tatgacattt tcacccatct cccatatcgg tttatcatca gcccttataa 3840

cctattatag cttgggggat atgtgttcaa aatatatggt taagtatggg gttatgaatc 3900

aacttgtaaa atatcatttt agctcagatg ttgagactgc acttttagtt gtggcataag 3960

ttgacacata caaccatgtg atacttaaga tgaattcatg aactaaagaa cctccatact 4020

tgaggtggtt ttccgggacc agaatatcaa aggactttat cctctattga tccaatatcc 4080

agcttgtcct gacctgtatt atttggagct tcactacatg ctggccaaca ttctagtaca 4140

agctcagcaa attttaggac acgagacatt ggtgaacttg tagctatcac tggattttac 4200

tttattttattggatttgct tagctattgg tgcagaatac aattattcg tgatttaatg 4260

tttatttagt tatgttcagc agttccatgc caccgtaaat agacatcacc atactcccct 4320

cgaaaagatt atgagtttcg aatttatcat gatgaaatat tctcaacaat tttcctgtac 4380

agggaatatt tatgctggtg tctggctact tcagacttcc atacgacatt ccaaagcctt 4440

tttggagata ccccatgcag tacatcagct tccatactg ggcactgcag gtaaattccg 4500

tttcacacat tggagcaacc cacataacact agcctccaag tctccaacct gaactttttg 4560

acaccaaact ccatacatct aacagggcca gtgccagaac gacatgaaag gcctcgtatt 4620

cgacaaccag taccccagatc aacccaagat ccccggggac ttcatcctga agtacatctt 4680

ccagatcaac gtggaccgga acaagtggat cgacttgtca gtcattttca gcatgatctt 4740

catctaccgc atcctcttct tcctcatgat caagatcaac gaggatgtgc tgccgtggat 4800

ccgtggccac atcgcgagga agagaatgca gaagaaaggc cctagtccca gtttcggcaa 4860

gacgccttcg ctgagaggct atgttgtgga tccagagctc ggttccaacg agggctagag 4920

gacgatgatg atctggttct tttgggtagc agaaataggg tgtttctccc cccggctctt 4980

ggtgctatat tgtatattta agatcaggga aaattaaact ggtatcatac gttttgaaac 5040

tgaagctaat tcgagtacag gtatacatac agcatgtaat gtgactatgt gagcaaagtg 5100

gcatcggaga gatgcccagt gacacatgac agattgttca gaatttattg ccttcgacac 5160

acgaagaatt gttggagcac taaagcgggt acggctcaca tgaccgtgta ttatcagatt 5220

gattttgatt gcttgctggt tacataaaat cggtactaca tattgacaga attgtaaatg 5280

ccccacataa atataaacta acaataagat ccatgccgta c 5321

<210> 14

<211> 2175

<212>DNA

<213> Sorghum (Sorghum bicolor)

<400> 14

atgagacggg ccggaggagg agacctgcag ggcgaccacg gcacgacgcc tcggtccgcg 60

gcggcagggc aggccatggt ggagctgcag gccaacgggt cggcggccgc cgccgggggc 120

gccatggtgg tgggtctcag cccgctcagc gagaccctgt ggcgggacag caaggcgctc 180

ccgccgggtg ccggccccgc cgcgctcatc ggcgacgtgt ccgccaggct cacgtggaag 240

gacctctgcg tcaccgtggc gctcggcccc ggcaagacgc agaccgtgct ggacgaactc 300

accgggtacg cggagcccgg ctcgctcacc gcgctcatgg gcccctccgg gtccggcaag 360

tccaccctgc tcgacgccct cgccggccgc ctcgccgcca acgccttcct ctccggcgac 420

gtgctcctca acggccgcaa ggccaagctc tccttcggcg ccgcggcgta cgtgacgcag 480

gacgacaacc tgatcgggac gctgacggtg cgcgagacga tcggctactc ggcgctgctg 540

cggctgccgg acaagatgcc gcgggaggac aagcgcgcgc tggtggaggg caccatcgtc 600

gagatggggc tccaggactg cgccgacacc gtcatcggga actggcacct ccgcggggtc 660

agcggcggcg agaagcgccg cgtcagcatc gcgctcgagc tcctcatgcg cccgcgcctc 720

ctcttcctcg acgagcccac cagcggcctc gacagctcct cggcgttctt cgtgacgcag 780

acgctgcgag gcctggccag ggacggcagg acagtgattg cttccatcca ccagcccagc 840

agcgaggtgt tcgagctctt cgacatgctc ttcctgctgt ccgggggcaa aaccgtctac 900

ttcggccaag catcgcaagc atgtgagttc tttgctcaag gcggtttccc ttgcccgcct 960

ctgaggaacc catcggacca tttcctgagg tgcgtcaact cggacttcga caaggtgaag 1020

gccaacactga aaggatcaat gaaggcaagg gctgagaggt ccgacgatcc actcgatagg 1080

atgactacat cagaagccat caggaaattg gttggatcct acagcaggtc ccaatactac 1140

tacgctgcga gggagaaagt taatgatac tcgagaatga aaggaactgt gctggattca 1200

ggtggcagcc aggctagctt cctgatgcag gcttgcaccc tgaccaagcg ctcgttcatc 1260

aacatgtcac gggactttgg gtattactgg ctcaggctcc tgatctatct cctcgtgact 1320

gtttgcatcg gcacaatcta cctggatgtt ggcactaaat acacatccat cctggccaga 1380

gcttcctgtt ctgcattcgt ctttgggttc gttacgttca tgtccattgg aggattccct 1440

tcgtttgttg aagaaatgaa ggtcttccaa cgggaacggc tgaatgggca ctatggtgtc 1500

gcggcatttg tcatcagtaa caccatctca gcaacgccat tcctgattct aatatgcttc 1560

ttgtcgggaa ctatatgcta ctttatggtg cgcctgcatc ctggtttcga acactatatc 1620

ttcttcgtct tgaacctgta tgccagcgtc acagtggtcg aaagcttaat gatggcaatt 1680

gccagtgtca ttcccaattt cctcatgggt atcatcatag gggctggaat tcagggaata 1740

tttatgctgg tgtctggcta cttcagactt ccatacgaca ttccaaagcc tttttggaga 1800

tacccccatgc agtacatcag cttccattac tgggcactgc agggccagtg ccagaacgac 1860

atgaaaggcc tcgtattcga caaccagtac ccagatcaac ccaagatccc cggggacttc 1920

atcctgaagt acatcttcca gatcaacgtg gaccggaaca agtggatcga cttgtcagtc 1980

attttcagca tgatcttcat ctaccgcatc ctcttcttcc tcatgatcaa gatcaacgag 2040

gatgtgctgc cgtggatccg tggccacatc gcgaggaaga gaatgcagaa gaaaggccct 2100

agtcccagtt tcggcaagac gccttcgctg agaggctatg ttgtggatcc agagctcggt 2160

tccaacgagg gctag 2175

<210> 15

<211> 724

<212> PRT

<213> Sorghum (Sorghum bicolor)

<400> 15

Met Arg Arg Ala Gly Gly Gly Asp Leu Gln Gly Asp His Gly Thr Thr

1 5 10 15

Pro Arg Ser Ala Ala Ala Gly Gln Ala Met Val Glu Leu Gln Ala Asn

20 25 30

Gly Ser Ala Ala Ala Ala Gly Gly Ala Met Val Val Gly Leu Ser Pro

35 40 45

Leu Ser Glu Thr Leu Trp Arg Asp Ser Lys Ala Leu Pro Pro Gly Ala

50 55 60

Gly Pro Ala Ala Leu Ile Gly Asp Val Ser Ala Arg Leu Thr Trp Lys

65 70 75 80

Asp Leu Cys Val Thr Val Ala Leu Gly Pro Gly Lys Thr Gln Thr Val

85 90 95

Leu Asp Glu Leu Thr Gly Tyr Ala Glu Pro Gly Ser Leu Thr Ala Leu

100 105 110

Met Gly Pro Ser Gly Ser Gly Lys Ser Thr Leu Leu Asp Ala Leu Ala

115 120 125

Gly Arg Leu Ala Ala Asn Ala Phe Leu Ser Gly Asp Val Leu Leu Asn

130 135 140

Gly Arg Lys Ala Lys Leu Ser Phe Gly Ala Ala Ala Tyr Val Thr Gln

145 150 155 160

Asp Asp Asn Leu Ile Gly Thr Leu Thr Val Arg Glu Thr Ile Gly Tyr

165 170 175

Ser Ala Leu Leu Arg Leu Pro Asp Lys Met Pro Arg Glu Asp Lys Arg

180 185 190

Ala Leu Val Glu Gly Thr Ile Val Glu Met Gly Leu Gln Asp Cys Ala

195 200 205

Asp Thr Val Ile Gly Asn Trp His Leu Arg Gly Val Ser Gly Gly Glu

210 215 220

Lys Arg Arg Val Ser Ile Ala Leu Glu Leu Leu Met Arg Pro Arg Leu

225 230 235 240

Leu Phe Leu Asp Glu Pro Thr Ser Gly Leu Asp Ser Ser Ser Ser Ala Phe

245 250 255

Phe Val Thr Gln Thr Leu Arg Gly Leu Ala Arg Asp Gly Arg Thr Val

260 265 270

Ile Ala Ser Ile His Gln Pro Ser Ser Glu Val Phe Glu Leu Phe Asp

275 280 285

Met Leu Phe Leu Leu Ser Gly Gly Lys Thr Val Tyr Phe Gly Gln Ala

290 295 300

Ser Gln Ala Cys Glu Phe Phe Ala Gln Gly Gly Phe Pro Cys Pro Pro

305 310 315 320

Leu Arg Asn Pro Ser Asp His Phe Leu Arg Cys Val Asn Ser Asp Phe

325 330 335

Asp Lys Val Lys Ala Thr Leu Lys Gly Ser Met Lys Ala Arg Ala Glu

340 345 350

Arg Ser Asp Asp Pro Leu Asp Arg Met Thr Thr Ser Glu Ala Ile Arg

355 360 365

Lys Leu Val Gly Ser Tyr Ser Arg Ser Gln Tyr Tyr Tyr Ala Ala Arg

370 375 380

Glu Lys Val Asn Asp Ile Ser Arg Met Lys Gly Thr Val Leu Asp Ser

385 390 395 400

Gly Gly Ser Gln Ala Ser Phe Leu Met Gln Ala Cys Thr Leu Thr Lys

405 410 415

Arg Ser Phe Ile Asn Met Ser Arg Asp Phe Gly Tyr Tyr Trp Leu Arg

420 425 430

Leu Leu Ile Tyr Leu Leu Val Thr Val Cys Ile Gly Thr Ile Tyr Leu

435 440 445

Asp Val Gly Thr Lys Tyr Thr Ser Ile Leu Ala Arg Ala Ser Cys Ser

450 455 460

Ala Phe Val Phe Gly Phe Val Thr Phe Met Ser Ile Gly Gly Phe Pro

465 470 475 480

Ser Phe Val Glu Glu Met Lys Val Phe Gln Arg Glu Arg Leu Asn Gly

485 490 495

His Tyr Gly Val Ala Ala Phe Val Ile Ser Asn Thr Ile Ser Ala Thr

500 505 510

Pro Phe Leu Ile Leu Ile Cys Phe Leu Ser Gly Thr Ile Cys Tyr Phe

515 520 525

Met Val Arg Leu His Pro Gly Phe Glu His Tyr Ile Phe Phe Val Leu

530 535 540

Asn Leu Tyr Ala Ser Val Thr Val Val Glu Ser Leu Met Met Ala Ile

545 550 555 560

Ala Ser Val Ile Pro Asn Phe Leu Met Gly Ile Ile Ile Gly Ala Gly

565 570 575

Ile Gln Gly Ile Phe Met Leu Val Ser Gly Tyr Phe Arg Leu Pro Tyr

580 585 590

Asp Ile Pro Lys Pro Phe Trp Arg Tyr Pro Met Gln Tyr Ile Ser Phe

595 600 605

His Tyr Trp Ala Leu Gln Gly Gln Cys Gln Asn Asp Met Lys Gly Leu

610 615 620

Val Phe Asp Asn Gln Tyr Pro Asp Gln Pro Lys Ile Pro Gly Asp Phe

625 630 635 640

Ile Leu Lys Tyr Ile Phe Gln Ile Asn Val Asp Arg Asn Lys Trp Ile

645 650 655

Asp Leu Ser Val Ile Phe Ser Met Ile Phe Ile Tyr Arg Ile Leu Phe

660 665 670

Phe Leu Met Ile Lys Ile Asn Glu Asp Val Leu Pro Trp Ile Arg Gly

675 680 685

His Ile Ala Arg Lys Arg Met Gln Lys Lys Gly Pro Ser Pro Ser Phe

690 695 700

Gly Lys Thr Pro Ser Leu Arg Gly Tyr Val Val Asp Pro Glu Leu Gly

705 710 715 720

Ser Asn Glu Gly

<210> 16

<211> 3616

<212>DNA

<213> Corn (Zea mays)

<400> 16

gagggagcgc attackctgc agcgcggaca cgcccggcac ggcaagccag ccactcttct 60

cgcgtgaagc ttagccattc ttgcggctgg gcaaggaaac tgctgaagcc actagccggc 120

tgcctgctct acggcctccg aaccgaacga ggtgcatcgt gaggaagcag cagcaggggg 180

gggggggggg ggggggcagt ggccggggcgg gatgacgggg agggaggccg tggcttcggc 240

ggaggcggag ctggatcagg actcggcggc ggcggcggcc gcggcgcccg tgctgctgag 300

cccgctcagc gagacgctgt ggcgcgacag ggcggcggcg gcggcgggcg gggtcctcgt 360

cggggacgtg tcggcgcgcc tggcgtggcg cgacctcacc gtgaccgtcg cgctcggcac 420

cggcgacacg caggccgtgc tgcagggcct cacggggcac gccgagcccg gcaccatcac 480

cgcgctcatg ggcccgtccg gctccggcaa gtccacgctg ctcgacgcgc tcgcgggccg 540

cctcgccgcc aacgccttcc tctccgggac cgtcctcctc aacgggcgca aggccaacct 600

ctccttcggc gccgcggtac gtgcgctcgt cctcgatcgg tcttctcctt tttctcctta 660

cgcatgacag aaagaggtgtgttttgttgttgtctataag ttgttcgtcg atctgatgaa 720

atagacatga caggacaggc tagctctcca cgcacgtttt tctgtgtgaa actgaaaccg 780

agcatttgtc tgtgactgtc agtctcctcc catgaacatg gagtggattt ggatttgtca 840

atctgctttt cagatggatc gtattgccaa tctcacctca cctggaacct tgttgccaat 900

aatactcagg cctacgtgac gcaagacgac aacctgatcg gcacgctgac ggtgagggag 960

acgatctcgt actcggcgcg actccggctc cccgacaaga tgccgaggga ggagaagcag 1020

gctctggtgg agggcaccat cgtggagatg gggctgcagg actgcgccga caccgtcgtc 1080

ggcaactggc acctgagggg gatcagcggc ggcgagaagc ggagggtcag catcgccctg 1140

gaaatactca tgcggcccag gctgctgttc ctggacgagc ctaccagtgg ccttgacagg 1200

tacttttata tatatgcgct tctgtttctt cgtcctatac tactactagt gatgaccgat 1260

gatgcaaaac aatgtgcagt gcttcggcgt tcttcgtgac gcagacgctg cgagggctgg 1320

caagggacgg ccggaccgtc atagcgtcgg tgcaccagcc gagcagcgag gttttcttgc 1380

tgttcgactg cctctacctt ctgtcagggg gcaggacagt ctacttcggg aaggcatcgg 1440

aggcctgcga ggtgaaggca ctggtctcta ctctctactc tactaccgat gcatatggct 1500

ggcatctgag agagccgtct atctgttgcg tgcacagttc ttcgcccagg ctggcttccc 1560

gtgcccgccg atgcgcaacc cgtcggacca tttcctccgg tgcataaact cggatttcga 1620

caaggtgaag gccactctca aagggtcgat gaaggcgagg gtgagtcgga tttcaccacc 1680

tcgctcctta gttttttttt tggctgttgt agcctatata catagcatct cctttcgctc 1740

gtgtagttcg aaaggtccaa cgacgacgac ccgctggaga agatgacgac gtcggaagcg 1800

atgaggaggc tcaccagcta ctaccagcac tcgcagtgcc acatcaacgc gcagcagaaa 1860

gtggacgaga tggcccggct cgtaagcgcc agctgctcca ttctcttgga cgatcgttca 1920

ccgctttcca acacaactga tgaaaggccg ggggggatcc atgtctgtct gtctgtcctc 1980

gcagaaagga acggtgctgg atccaggagg agggagccag gccagcttcc tgatgcaggc 2040

gttcacgctc acgaagcgct cgttcgtcaa catgtccagg gacttcggct actactggct 2100

caggctcatc atctacatcg tcgtcacgct ttgcatcggg actatatacc tcaacgttgg 2160

gaccggatac agctctatcc tggtaattat taacacgcaa catctatcgt cgtcttttgt 2220

tgggttaatt gggcactgaa tgaataataa tgcgtctact tgcttgttct gaatctgact 2280

gatgaaaact actatactac taggcccggg gcgcgtgtgc ttcgttcatc ttcggcttcg 2340

tcacgttcat gtccatcgga gggtttccgt ctttcgtgga ggacatgaag gtctgtctct 2400

gtcatcggag ggtttcctat gctcagggcc tcaggctcag cttattaatt caattcccgt 2460

ttctttataa ttctccaggt gttccagagg gagaggctga acgggcacta cggcgtgctg 2520

gcgttcgtcg tcagcaacac ggtctcggcg atgccgttcc tggtcctcat caccttcgtg 2580

tcgggcacgc tgtgctactt catggtgcgc ctgcacccgg gcttcacgca ctacctcttc 2640

ttcgtgctgg cgctctacgc cagcgtcacc gtcgtcgaga gcctcatgat ggccatcgcc 2700

agcgtcatcc ccaacttcct catgggcatc ataatcggcg cagggatcca ggtaattaaa 2760

ctgagactga gagactgaga ctgagactga gggttcccta atggtcgtcg tggacacaca 2820

caggggatct tcatgctggt gtcgggttac ttcaggcttc cgcacgacat ccccaagccc 2880

ttctggaggt acccccatgtc atacatcagc ttccactact gggcgctgca ggggcagtac 2940

cagaacgacc tcaggggcct gctgttcgac aaccaggacg acgagctgcc caggatcccg 3000

ggggagttca tcctggagac cgtgttccag atcgacgtcg cccgctccaa gtggctggac 3060

ctggccgtgc tcttcagcat gatcgtcatc taccgcctgc tcttcttcct catgatcaag 3120

gccagcgagg acgtcacccc ctggctgcgc cgctacgtcg cgcggcgccg ggtgcggaac 3180

cgtcgccacc gcaagccgga cctgcccgcc cggacgccgt cgctccgcgg gtacgtcgtc 3240

gacgcggcca gcctgccgga cgaccacccg tgacgaaaga gcgcgagaga aaacgtacgg 3300

gcagggatgt catttcatat catatcacac agaggctaaa ccgggatcgag ctgataataa 3360

cacactcact cactagtact gcaataatat gatgttaagt tggtatagca accgcttaag 3420

tgcccgttcg gttcttcctg aatgctttac tggattggtt ccggattgga atagttcaca 3480

aatttgtatt agctcgatga gttggaatga ttccggcctt caatctggac taaacgaaca 3540

gagctgtaat ggagtgttgc ttgatgcact gctaagatgt acacgtatat gaagagcatg 3600

tgaccatcag ttctac 3616

<210> 17

<211> 2127

<212>DNA

<213> Corn (Zea mays)

<400> 17

atgacgggga gggaggccgt ggcggcggcg gaggcggagc tggatcagga ctcggcggcg 60

gcggcggcgc ccgtgctgct gagcccgctc agcgagacgc tgtggcgcga cagggcggcg 120

gcggcggcgg gcggggtcct cgtcggggac gtgtcggcgc gcctggcgtg gcgcgacctc 180

accgtgaccg tcgcgctcgg ctccggcgac acgcaggccg tgctgcaggg cctcacgggg 240

cacgccgagc ccggcaccat caccgcgctc atgggcccgt ccggctccgg caagtccacg 300

ctgctcgacg cgctcgccgg ccgcctcgcc gccaacgcct tcctctccgg gaccgtcctc 360

ctcaacgggc gcaaggccaa cctctccttc ggcgccgcgg cctacgtgac gcaagacgac 420

aacctgatcg gcacgctgac ggtgagggag acgatctcgt actcggcgcg actccggctc 480

cccgacaaga tgccgaggga ggagaagcag gctctggtgg agggcaccat cgtggagatg 540

gggctgcagg actgcgccga caccgtcgtc ggcaactggc acctgagggg gatcagcggc 600

ggcgagaagc ggagggtcag catcgccctg gaaatactca tgcggcccag gctgctgttc 660

ctggacgagc ctaccagtgg ccttgacagt gcttcggcgt tcttcgtgac gcagacgctg 720

cgagggctgg caagggacgg ccggaccgtc atagcgtcgg tgcaccagcc gagcagcgag 780

gttttcttgc tgttcgactg cctctacctt ctgtcggggg gcaggacagt ctacttcggg 840

aaggcatcgg aggcctgcga gttcttcgcg caggctggct tcccgtgccc gccgatgcgc 900

aacccgtcgg accatttcct ccggtgcata aactcggatt tcgacaaggt gaaggccact 960

ctcaaagggt ccatgaaggc gaagttcgaa aggtccaacg acgacgaccc gctggagaag 1020

atgacgacgt cggaagcgat gaggaggctg accagctact accagcactc gcagtgccac 1080

atcaacgcgc agcagaaagt ggacgagatg gcccggctca aaggaacggt gctggatcca 1140

ggaggaggga gccaggccag cttcctgatg caggcgttca cgctcacgaa gcgctcgttc 1200

gtcaacatgt ccagggactt cggctactac tggctcaggc tcatcatcta catcgtcgtc 1260

acgctttgca tcgggactat atacctcaac gttgggaccg gatacagctc tatcctggcc 1320

cggggcgcgt gtgcttcgtt catcttcggc ttcgtcacgt tcatgtccat cggagggttt 1380

ccgtctttcg tggaggacat gaaggtgttc cagagggaga ggctgaacgg gcactacggc 1440

gtgctggcgt tcgtcgtcag caacacggtc tcggcgatgc cgttcctggt cctcatcacc 1500

ttcgtgtcgg gcacgctgtg ctacttcatg gtgcgcctgc acccgggctt cacgcactac 1560

ctcttcttcg tgctggcgct ctacgccagc gtcaccgtcg tcgagagcct catgatggcc 1620

atcgccagcg tcatccccaa cttcctcatg ggcatcataa tcggcgcagg gatccagggg 1680

atcttcatgc tggtgtcggg ttacttcagg cttccgcacg acatccccaa gcccttctgg 1740

aggtacccca tgtcatacat cagcttccac tactgggcgc tgcaggggca gtaccagaac 1800

gacctcaggg gcctgctgtt cgacaaccag gacgacgagc tgcccaggat cccgggggag 1860

ttcatcctgg agaccgtgtt ccagatcgac gtcgcccgct ccaagtggct ggacctggcc 1920

gtgctcttca gcatgatcgt catctaccgc ctgctcttct tcctcatgat caaggccagc 1980

gaggacgtca ccccctggct gcgccgctac gtcgcgcggc gccgggtgcg gaaccgtcgc 2040

caccgcaagc cggacctgcc cgcccggacg ccgtcgctcc gcgggtacgt cgtcgacgcg 2100

gccagcctgc cggacgacca cccgtga 2127

<210> 18

<211> 708

<212> PRT

<213> Corn (Zea mays)

<400> 18

Met Thr Gly Arg Glu Ala Val Ala Ala Ala Glu Ala Glu Leu Asp Gln

1 5 10 15

Asp Ser Ala Ala Ala Ala Ala Pro Val Leu Leu Ser Pro Leu Ser Glu

20 25 30

Thr Leu Trp Arg Asp Arg Ala Ala Ala Ala Ala Gly Gly Val Leu Val

35 40 45

Gly Asp Val Ser Ala Arg Leu Ala Trp Arg Asp Leu Thr Val Thr Val

50 55 60

Ala Leu Gly Ser Gly Asp Thr Gln Ala Val Leu Gln Gly Leu Thr Gly

65 70 75 80

His Ala Glu Pro Gly Thr Ile Thr Ala Leu Met Gly Pro Ser Gly Ser

85 90 95

Gly Lys Ser Thr Leu Leu Asp Ala Leu Ala Gly Arg Leu Ala Ala Asn

100 105 110

Ala Phe Leu Ser Gly Thr Val Leu Leu Asn Gly Arg Lys Ala Asn Leu

115 120 125

Ser Phe Gly Ala Ala Ala Tyr Val Thr Gln Asp Asp Asn Leu Ile Gly

130 135 140

Thr Leu Thr Val Arg Glu Thr Ile Ser Tyr Ser Ala Arg Leu Arg Leu

145 150 155 160

Pro Asp Lys Met Pro Arg Glu Glu Lys Gln Ala Leu Val Glu Gly Thr

165 170 175

Ile Val Glu Met Gly Leu Gln Asp Cys Ala Asp Thr Val Val Gly Asn

180 185 190

Trp His Leu Arg Gly Ile Ser Gly Gly Glu Lys Arg Arg Val Ser Ile

195 200 205

Ala Leu Glu Ile Leu Met Arg Pro Arg Leu Leu Phe Leu Asp Glu Pro

210 215 220

Thr Ser Gly Leu Asp Ser Ala Ser Ala Phe Phe Val Thr Gln Thr Leu

225 230 235 240

Arg Gly Leu Ala Arg Asp Gly Arg Thr Val Ile Ala Ser Val His Gln

245 250 255

Pro Ser Ser Glu Val Phe Leu Leu Phe Asp Cys Leu Tyr Leu Leu Ser

260 265 270

Gly Gly Arg Thr Val Tyr Phe Gly Lys Ala Ser Glu Ala Cys Glu Phe

275 280 285

Phe Ala Gln Ala Gly Phe Pro Cys Pro Pro Met Arg Asn Pro Ser Asp

290 295 300

His Phe Leu Arg Cys Ile Asn Ser Asp Phe Asp Lys Val Lys Ala Thr

305 310 315 320

Leu Lys Gly Ser Met Lys Ala Lys Phe Glu Arg Ser Asn Asp Asp Asp

325 330 335

Pro Leu Glu Lys Met Thr Thr Ser Glu Ala Met Arg Arg Leu Thr Ser

340 345 350

Tyr Tyr Gln His Ser Gln Cys His Ile Asn Ala Gln Gln Lys Val Asp

355 360 365

Glu Met Ala Arg Leu Lys Gly Thr Val Leu Asp Pro Gly Gly Gly Ser

370 375 380

Gln Ala Ser Phe Leu Met Gln Ala Phe Thr Leu Thr Lys Arg Ser Phe

385 390 395 400

Val Asn Met Ser Arg Asp Phe Gly Tyr Tyr Trp Leu Arg Leu Ile Ile

405 410 415

Tyr Ile Val Val Thr Leu Cys Ile Gly Thr Ile Tyr Leu Asn Val Gly

420 425 430

Thr Gly Tyr Ser Ser Ile Leu Ala Arg Gly Ala Cys Ala Ser Phe Ile

435 440 445

Phe Gly Phe Val Thr Phe Met Ser Ile Gly Gly Phe Pro Ser Phe Val

450 455 460

Glu Asp Met Lys Val Phe Gln Arg Glu Arg Leu Asn Gly His Tyr Gly

465 470 475 480

Val Leu Ala Phe Val Val Ser Asn Thr Val Ser Ala Met Pro Phe Leu

485 490 495

Val Leu Ile Thr Phe Val Ser Gly Thr Leu Cys Tyr Phe Met Val Arg

500 505 510

Leu His Pro Gly Phe Thr His Tyr Leu Phe Phe Val Leu Ala Leu Tyr

515 520 525

Ala Ser Val Thr Val Val Glu Ser Leu Met Met Ala Ile Ala Ser Val

530 535 540

Ile Pro Asn Phe Leu Met Gly Ile Ile Ile Gly Ala Gly Ile Gln Gly

545 550 555 560

Ile Phe Met Leu Val Ser Gly Tyr Phe Arg Leu Pro His Asp Ile Pro

565 570 575

Lys Pro Phe Trp Arg Tyr Pro Met Ser Tyr Ile Ser Phe His Tyr Trp

580 585 590

Ala Leu Gln Gly Gln Tyr Gln Asn Asp Leu Arg Gly Leu Leu Phe Asp

595 600 605

Asn Gln Asp Asp Glu Leu Pro Arg Ile Pro Gly Glu Phe Ile Leu Glu

610 615 620

Thr Val Phe Gln Ile Asp Val Ala Arg Ser Lys Trp Leu Asp Leu Ala

625 630 635 640

Val Leu Phe Ser Met Ile Val Ile Tyr Arg Leu Leu Phe Phe Leu Met

645 650 655

Ile Lys Ala Ser Glu Asp Val Thr Pro Trp Leu Arg Arg Tyr Val Ala

660 665 670

Arg Arg Arg Val Arg Asn Arg Arg His Arg Lys Pro Asp Leu Pro Ala

675 680 685

Arg Thr Pro Ser Leu Arg Gly Tyr Val Val Asp Ala Ala Ser Leu Pro

690 695 700

Asp Asp His Pro

705

<210> 19

<211> 2154

<212>DNA

<213> Millet (Setaria italica)

<400> 19

atgaggaagg ggggagcggc tgtcggggcg gcgatggtgg ggagggaggc ggcggtcgcg 60

gcggtggctg cggagctgga ggactcggcg gggaaggcgg cgggggcggc ggcgcccgcg 120

ctgagcccgc tcagcgagac gctgtggcgg gacagggcgg cggcgggcgg ggtcctcctc 180

ggtgacgtgt cggcgcgact ggcctggcgg gacctcgccg tcaccgtcgc gctcggcacc 240

ggcgacacgc aggccgtgct gcagggcctc acgggccacg ccgagccggg caccatcacg 300

gcgctcatgg ggccctccgg ctccggcaag tccacgctcc tcgacgcgct cgccgggcgg 360

ctcgccgcca acgccttcct ctccgggacc gtcctcctca acggacgcaa ggccaacctc 420

tccttcggcg ccgcggccta tgtgacgcaa gatgacaacc ttatcggcac attgacggtt 480

cgggagacga tctcatactc ggcgcgcctc cgtctccctg acaagatgcc ctgggaggag 540

aagcaggccc tggtagaggg caccatcatc gagatggggc tgcaggactg tgccgacacg 600

gtcgtcggca actggcacct gaggggcatc agtggtggtg aaaagaggag ggtcagcatt 660

gcccttgaga tactgatgag gcccaggctg ctcttcctgg acgagccaac cagtggtctc 720

gatagcgctt cagcattctt cgtgacacag acactgcgtg ggctggcgag ggacggccgg 780

acggtcatcg cgtccgtgca tcagccgagc agtgaggtgt tcacgctctt tgactgtctg 840

tatcttctat cgggagggaa aactgtctac tttgggaagg cctctgaggc ttgtgagttc 900

ttcgcccaag ctggtttccc gtgcccacca atgcgcaacc cctcggatca ttttctcagg 960

tgcataaatt cggattttga taaggtgaag gccactctga aaggatcaat gaagacgaga 1020

tttgaaaggt ctgatgatcc gctcgagaaa atcacaactt cggaggcaat gaggaggctg 1080

atcaactact accagcactc acagtaccac atcaatgcgc agcagaaagt agatgagatg 1140

gcccgggtta aaggaacagt gctggactca ggagggagcc aagctagttt cgcaatgcag 1200

gcgttcacac tcacgaagcg atcattcatc aacatgtcaa gggattttgg atactactgg 1260

ttgaggctta tcatctacat cgtcgtgaca ctctgcattg ggactatcta cctaaatgtt 1320

ggcactggat acagctccat tctggctcgg ggcgcatgtg cttccttcgt atttggcttt 1380

gtcacattca tgtcgattgg agggtttcca tcgtttgtcg aggacatgaa ggtctttcaa 1440

aggggagaggc tcaatgggca ctacggtgtg ctggcgtttg tcatcagcaa cacgctgtca 1500

gcgatgccgt tcctgatcct gatcaccttc gtgtcgggca cgctgtgcta cttcatggtg 1560

cgcctccacc cgggcttcat gcactacctc ttcttcgtcc tggccctcta tgccagcgtc 1620

actgttgtcg agagcctgat gatggccatt gccagcgtga tccccaactt cctcatgggc 1680

atcattatcg gcgcagggat tcagggcata ttcatgcttg tatcaggcta cttcaggctt 1740

ccacatgata tcccaaagcc tttctggagg tacccccatgt catatatcag cttccactac 1800

tgggcattgc agggccagta ccagaacgat ctcaagggcc tggtgttcga caaccaggac 1860

gacgagctgc ccaagatccc cggggagtac atcctggaga acgtgttcca gatcgacgtg 1920

aaccggtcca agtggctgga cctggccgtg ctgttcagca tgatcgtcat ctaccgcctc 1980

ctcttcttcg tcatgatcaa gatcagcgag gacgtgaccc cctgggtgcg cggctacatc 2040

gcgcggcggc gggtgcagaa ccggcggcag cgcaaggtgg agctggccgc ccggtcgccg 2100

tcgctccgcg ggtacgtggt ggacgtggcc agcctgccgg ccgaccacccc ctga 2154

<210> 20

<211> 717

<212> PRT

<213> Millet (Setaria italica)

<400> 20

Met Arg Lys Gly Gly Ala Ala Val Gly Ala Ala Met Val Gly Arg Glu

1 5 10 15

Ala Ala Val Ala Ala Val Ala Ala Glu Leu Glu Asp Ser Ala Gly Lys

20 25 30

Ala Ala Gly Ala Ala Ala Pro Ala Leu Ser Pro Leu Ser Glu Thr Leu

35 40 45

Trp Arg Asp Arg Ala Ala Ala Gly Gly Val Leu Leu Gly Asp Val Ser

50 55 60

Ala Arg Leu Ala Trp Arg Asp Leu Ala Val Thr Val Ala Leu Gly Thr

65 70 75 80

Gly Asp Thr Gln Ala Val Leu Gln Gly Leu Thr Gly His Ala Glu Pro

85 90 95

Gly Thr Ile Thr Ala Leu Met Gly Pro Ser Gly Ser Gly Lys Ser Thr

100 105 110

Leu Leu Asp Ala Leu Ala Gly Arg Leu Ala Ala Asn Ala Phe Leu Ser

115 120 125

Gly Thr Val Leu Leu Asn Gly Arg Lys Ala Asn Leu Ser Phe Gly Ala

130 135 140

Ala Ala Tyr Val Thr Gln Asp Asp Asn Leu Ile Gly Thr Leu Thr Val

145 150 155 160

Arg Glu Thr Ile Ser Tyr Ser Ala Arg Leu Arg Leu Pro Asp Lys Met

165 170 175

Pro Trp Glu Glu Lys Gln Ala Leu Val Glu Gly Thr Ile Ile Glu Met

180 185 190

Gly Leu Gln Asp Cys Ala Asp Thr Val Val Gly Asn Trp His Leu Arg

195 200 205

Gly Ile Ser Gly Gly Glu Lys Arg Arg Val Ser Ile Ala Leu Glu Ile

210 215 220

Leu Met Arg Pro Arg Leu Leu Phe Leu Asp Glu Pro Thr Ser Gly Leu

225 230 235 240

Asp Ser Ala Ser Ala Phe Phe Val Thr Gln Thr Leu Arg Gly Leu Ala

245 250 255

Arg Asp Gly Arg Thr Val Ile Ala Ser Val His Gln Pro Ser Ser Glu

260 265 270

Val Phe Thr Leu Phe Asp Cys Leu Tyr Leu Leu Ser Gly Gly Lys Thr

275 280 285

Val Tyr Phe Gly Lys Ala Ser Glu Ala Cys Glu Phe Phe Ala Gln Ala

290 295 300

Gly Phe Pro Cys Pro Pro Met Arg Asn Pro Ser Asp His Phe Leu Arg

305 310 315 320

Cys Ile Asn Ser Asp Phe Asp Lys Val Lys Ala Thr Leu Lys Gly Ser

325 330 335

Met Lys Thr Arg Phe Glu Arg Ser Asp Asp Pro Leu Glu Lys Ile Thr

340 345 350

Thr Ser Glu Ala Met Arg Arg Leu Ile Asn Tyr Tyr Gln His Ser Gln

355 360 365

Tyr His Ile Asn Ala Gln Gln Lys Val Asp Glu Met Ala Arg Val Lys

370 375 380

Gly Thr Val Leu Asp Ser Gly Gly Ser Gln Ala Ser Phe Ala Met Gln

385 390 395 400

Ala Phe Thr Leu Thr Lys Arg Ser Phe Ile Asn Met Ser Arg Asp Phe

405 410 415

Gly Tyr Tyr Trp Leu Arg Leu Ile Ile Tyr Ile Val Val Thr Leu Cys

420 425 430

Ile Gly Thr Ile Tyr Leu Asn Val Gly Thr Gly Tyr Ser Ser Ile Leu

435 440 445

Ala Arg Gly Ala Cys Ala Ser Phe Val Phe Gly Phe Val Thr Phe Met

450 455 460

Ser Ile Gly Gly Phe Pro Ser Phe Val Glu Asp Met Lys Val Phe Gln

465 470 475 480

Arg Glu Arg Leu Asn Gly His Tyr Gly Val Leu Ala Phe Val Ile Ser

485 490 495

Asn Thr Leu Ser Ala Met Pro Phe Leu Ile Leu Ile Thr Phe Val Ser

500 505 510

Gly Thr Leu Cys Tyr Phe Met Val Arg Leu His Pro Gly Phe Met His

515 520 525

Tyr Leu Phe Phe Val Leu Ala Leu Tyr Ala Ser Val Thr Val Val Glu

530 535 540

Ser Leu Met Met Ala Ile Ala Ser Val Ile Pro Asn Phe Leu Met Gly

545 550 555 560

Ile Ile Ile Gly Ala Gly Ile Gln Gly Ile Phe Met Leu Val Ser Gly

565 570 575

Tyr Phe Arg Leu Pro His Asp Ile Pro Lys Pro Phe Trp Arg Tyr Pro

580 585 590

Met Ser Tyr Ile Ser Phe His Tyr Trp Ala Leu Gln Gly Gln Tyr Gln

595 600 605

Asn Asp Leu Lys Gly Leu Val Phe Asp Asn Gln Asp Asp Glu Leu Pro

610 615 620

Lys Ile Pro Gly Glu Tyr Ile Leu Glu Asn Val Phe Gln Ile Asp Val

625 630 635 640

Asn Arg Ser Lys Trp Leu Asp Leu Ala Val Leu Phe Ser Met Ile Val

645 650 655

Ile Tyr Arg Leu Leu Phe Phe Val Met Ile Lys Ile Ser Glu Asp Val

660 665 670

Thr Pro Trp Val Arg Gly Tyr Ile Ala Arg Arg Arg Val Gln Asn Arg

675 680 685

Arg Gln Arg Lys Val Glu Leu Ala Ala Arg Ser Pro Ser Leu Arg Gly

690 695 700

Tyr Val Val Asp Val Ala Ser Leu Pro Ala Asp His Pro

705 710 715

<210> 21

<211> 3762

<212>DNA

<213> Brachypodium distachyon

<400> 21

atgaagggag ctgaggcggc ggcggcggcg aaggcggagc tggaggacgt ggggaaggcg 60

gcggtccggg cggcggcgcc cccgctgagc ccgctgagcg agacgctgtg gcgggacaag 120

gcggggctcg tcttgctcgg ggacgtgtcg gcgcggctag cgtggcggga cctcacggtg 180

accgtgccgg cggcgggcgg cggcgggacg caggcggtgc tggaggcgct gacggggtac 240

gcggagccgg gcaccatgac ggcgctcatg gggccctccg gctccggcaa gtccactctc 300

ctcgacgcgc tcgccggccg cctcgccgcc aacgcgtttc tatctggcac cgtcctcctc 360

aacgggcgca aggccaacct ctccttcggc gccgcggtac gtaccctgca tatgcctcgc 420

cgtccacggc tagctagcta ctgtaactgg ctgccgctct ctcgctcgat ctcttaatta 480

agctagcttc cctgctagta ggtatactgg cgtactgatc tttagacggt agagtgtact 540

ctgtagtctg tagtgtacat caacaagcaa caagaggtcc ggttctatta aagattcatg 600

catttttcat caacctttta aaagtacatt tttttccctc tacatacata tatccaacaa 660

agaatcctgt gcaatcgaac atatttgttc gtcaatccca tgactttcta ggtagatttg 720

ggcatacaac acggtagcag tttggtgatg gtatacgact agctagcccg ttttactata 780

atcagtctgt ttttcttctt cttttacttc cattccttat ctgtgtttct ccccatgggc 840

catcttgaat tgtgattgaa tctctctgcg ccacctgaga tggcaaccat gcgatgctca 900

caaaccagaa agaagtaatg gaccatgtgc cgcgtggcag gttgtttgga accttgctgg 960

ttcttggctt ctgcctgctc agcttagctg catttgctat ttgttggagg ttatctccta 1020

attgactgca catgaactgc ttctgtctgg acacggatta gcaagttcct agtctcaatt 1080

tgagatctca catcttcttt aatctgggcg tcttgtttgg tgatcgaatt tgacacccat 1140

tttttctaga ggatttttag ttcccatttt ggttggttgg taaagggatg caaggcactt 1200

tttttgtacc tgataagtgg caactggaag cagatgggtt atggttcacc agtttttttt 1260

tcctcaggag tctgctcata ggaaactcat gacaccttca gaacatagta ttctttttgt 1320

ctgcagtgtg aaaattcagt ttctgtttgt taacgagaag tgtgtgtgca tgcgtatgcg 1380

tgtgtctatg ttacctgaaa ccattaatca gatgcatgca atttgtgatt cgggcaggca 1440

tatgtgacgc aagatgacaa tctgatgggc acactgacgg tgagggagac gatctcatac 1500

tcggcgagcc ttcggctccc tgacaagatg cccatggaag agaagcgtga cctggtggag 1560

ggcacaatcg tggagatggg gctccaggac tgcgccgaca cggtcatcgg caattggcac 1620

ctcagggggg tcagcggcgg tgagaagaga agggtcagca tcgccctgga gatcctcatg 1680

aggcctaggc tgctcttcct ggatgagccc accagtggtc ttgacaggta tatatggcat 1740

cttcacaatt tcagtcatgc aattcttttt attcagttgt ttgtaaatat tttgctcgca 1800

tacaagttcg ataatgatac atatctgaat tctgaaaact atgcttgtcg tgcagcgctt 1860

cagcgttctt cgtgacgcag acgctccgcg ggctggcgag ggatgggcgg acagtgatcg 1920

cgtcgatcca tcagccgagc agcgaggtct tccagctctt cgatcgcctg tatcttcttt 1980

cagggggcaa aacagtctac tttgggcagg cgtctgaggc ttgtgaggta ctctgtagct 2040

taatcttttt atctgtattt cttcaatgcc aaattgccaa tgccaatggg gtctctcaag 2100

cagtctgaat atgcaatgca gttctttgcc caagctggct tcccatgccc agcactacgc 2160

aacccgtcgg atcatttcct ccgctgcata aacgcagact tcgacaaggt gaaggccacc 2220

ctgaaaggat caatgaagat gagagtaagt ttcttcagcc actatttcac tgatgaatct 2280

caattagctt tctgatggac cttaattagt ttgagaggtc cgacgatccg ctagagcgca 2340

tcacgacctg cgaggcaatc agaaagttat tcagccacta ccagcactcg caggactacc 2400

tcgccacccg gcagaaagtg gacgagatgg cacgggtcgt aagtgcagct gatccatttt 2460

tcttcagaaa cctacctgca atgacaagta aattacgatg gttcattttg catctgcaga 2520

aaggaacggt cctggacgca gggggcagcc aggccagctt cgggatgcaa gcctgcacgc 2580

tgacgaagcg gtccttcgtg aacatgacga gggacttcgg gtactactgg ctgcggctgg 2640

tgatctacgt ggtggtcacc gtgtgcatcg gctccatcta cctcaacgtg ggcaccaagt 2700

acagctccat cctggcgcgg ggcgcgtgcg cgtccttcat cttcggcttc gtcaccttca 2760

tgtccatcgg gggcttcccc tccttcgtgg aggacatgaa ggtgttccag cgggagcggc 2820

tcaacggcca ctacggcgtg gccgcgttcg tgctcggcaa cacggcgtcc gcggcgccgt 2880

tcctgctgct catcaccgtc tgctccggga cgctatgcta cttcatggtg gggctacacc 2940

cggggctcag ccattatgtt ttcttcctgc tctgcctcta tgccagcgtc accgtcgtcg 3000

agagcctcat gatggccatc gccagcgtcg tcccgaattt tctcatgggc atcatcaccg 3060

gcgccggaat tcaaggggtg ttcatgctcg tctcggggta tttccgcctc ccccacgaca 3120

tccccaagcc cttctggcgt tacccccatgt catacatcag cttccactac tgggccctgc 3180

aggtaacatc cagatcgatt tttccatctt tcctctgaat gaagcgtcaa gatcctgcat 3240

gcacgcatgg tgtttgatgg attgattttg caggggcagt accagaacga tttggtgggg 3300

ctcatgttcg acaaccagag cgagttgctg cccaagatcc cgggggagta cgtgctggag 3360

aacgtgttcc agatcgacgt ggggaggtca aagtggctgg acttgtccgt gcttttcgcc 3420

atgatcgtcg tctaccgcct gctcttcttc gccatgatca aggtcagcga ggacgtgacg 3480

ccgtggctgc gccggtacat cgccaggagg agggtgcagc gcaggcgagg ccgggcccgg 3540

gacgcggcgg cggagctcca agttggccgc tcgccgtcgc tgcgcgggta cgtcgtcgac 3600

gacgacctgc cggccgatca tccgtgacag tatgagctca agaagaaagg ttaattggtt 3660

ggacagacga aattaacgct aaaataactg gacgataaaa cagtctgata tgaatgatgt 3720

actcgtatgt gtgttagcta gggcatctcc gatttgttct ct 3762

<210> 22

<211> 2127

<212>DNA

<213> Brachypodium distachyon

<400> 22

atgaagggag ctgaggcggc ggcggcggcg aaggcggagc tggaggacgt ggggaaggcg 60

gcggtccggg cggcggcgcc cccgctgagc ccgctgagcg agacgctgtg gcgggacaag 120

gcggggctcg tcttgctcgg ggacgtgtcg gcgcggctag cgtggcggga cctcacggtg 180

accgtgccgg cggcgggcgg cggcgggacg caggcggtgc tggaggcgct gacggggtac 240

gcggagccgg gcaccatgac ggcgctcatg gggccctccg gctccggcaa gtccactctc 300

ctcgacgcgc tcgccggccg cctcgccgcc aacgcgtttc tatctggcac cgtcctcctc 360

aacgggcgca aggccaacct ctccttcggc gccgcggcat atgtgacgca agatgacaat 420

ctgatgggca cactgacggt gagggagacg atctcatact cggcgagcct tcggctccct 480

gacaagatgc ccatggaaga gaagcgtgac ctggtggagg gcacaatcgt ggagatgggg 540

ctccaggact gcgccgacac ggtcatcggc aattggcacc tcaggggggt cagcggcggt 600

gagaagagaa gggtcagcat cgccctggag atcctcatga ggcctaggct gctcttcctg 660

gatgagccca ccagtggtct tgacagcgct tcagcgttct tcgtgacgca gacgctccgc 720

gggctggcga gggatgggcg gacagtgatc gcgtcgatcc atcagccgag cagcgaggtc 780

ttccagctct tcgatcgcct gtatcttctt tcagggggca aaacagtcta ctttgggcag 840

gcgtctgagg cttgtgagtt ctttgcccaa gctggcttcc catgcccagc actacgcaac 900

ccgtcggatc atttcctccg ctgcataaac gcagacttcg acaaggtgaa ggccaccctg 960

aaaggatcaa tgaagatgag atttgagagg tccgacgatc cgctagagcg catcacgacc 1020

tgcgaggcaa tcagaaagtt attcagccac taccagcact cgcaggacta cctcgccacc 1080

cggcagaaag tggacgagat ggcacgggtc aaaggaacgg tcctggacgc aggggggcagc 1140

caggccagct tcgggatgca agcctgcacg ctgacgaagc ggtccttcgt gaacatgacg 1200

agggacttcg ggtactactg gctgcggctg gtgatctacg tggtggtcac cgtgtgcatc 1260

ggctccatct acctcaacgt gggcaccaag tacagctcca tcctggcgcg gggcgcgtgc 1320

gcgtccttca tcttcggctt cgtcaccttc atgtccatcg ggggcttccc ctccttcgtg 1380

gaggacatga aggtgttcca gcgggagcgg ctcaacggcc actacggcgt ggccgcgttc 1440

gtgctcggca acacggcgtc cgcggcgccg ttcctgctgc tcatcaccgt ctgctccggg 1500

acgctatgct acttcatggt ggggctacac ccggggctca gccattatgt tttcttcctg 1560

ctctgcctct atgccagcgt caccgtcgtc gagagcctca tgatggccat cgccagcgtc 1620

gtcccgaatt ttctcatggg catcatcacc ggcgccggaa ttcaaggggt gttcatgctc 1680

gtctcggggt atttccgcct cccccacgac atccccaagc ccttctggcg ttaccccatg 1740

tcatacatca gcttccacta ctgggccctg caggggcagt accagaacga tttggtgggg 1800

ctcatgttcg acaaccagag cgagttgctg cccaagatcc cgggggagta cgtgctggag 1860

aacgtgttcc agatcgacgt ggggaggtca aagtggctgg acttgtccgt gcttttcgcc 1920

atgatcgtcg tctaccgcct gctcttcttc gccatgatca aggtcagcga ggacgtgacg 1980

ccgtggctgc gccggtacat cgccaggagg agggtgcagc gcaggcgagg ccgggcccgg 2040

gacgcggcgg cggagctcca agttggccgc tcgccgtcgc tgcgcgggta cgtcgtcgac 2100

gacgacctgc cggccgatca tccgtga 2127

<210> 23

<211> 708

<212> PRT

<213> Brachypodium distachyon

<400> 23

Met Lys Gly Ala Glu Ala Ala Ala Ala Ala Ala Lys Ala Glu Leu Glu Asp

1 5 10 15

Val Gly Lys Ala Ala Val Arg Ala Ala Ala Pro Pro Leu Ser Pro Leu

20 25 30

Ser Glu Thr Leu Trp Arg Asp Lys Ala Gly Leu Val Leu Leu Gly Asp

35 40 45

Val Ser Ala Arg Leu Ala Trp Arg Asp Leu Thr Val Thr Val Pro Ala

50 55 60

Ala Gly Gly Gly Gly Thr Gln Ala Val Leu Glu Ala Leu Thr Gly Tyr

65 70 75 80

Ala Glu Pro Gly Thr Met Thr Ala Leu Met Gly Pro Ser Gly Ser Gly

85 90 95

Lys Ser Thr Leu Leu Asp Ala Leu Ala Gly Arg Leu Ala Ala Asn Ala

100 105 110

Phe Leu Ser Gly Thr Val Leu Leu Asn Gly Arg Lys Ala Asn Leu Ser

115 120 125

Phe Gly Ala Ala Ala Tyr Val Thr Gln Asp Asp Asn Leu Met Gly Thr

130 135 140

Leu Thr Val Arg Glu Thr Ile Ser Tyr Ser Ala Ser Leu Arg Leu Pro

145 150 155 160

Asp Lys Met Pro Met Glu Glu Lys Arg Asp Leu Val Glu Gly Thr Ile

165 170 175

Val Glu Met Gly Leu Gln Asp Cys Ala Asp Thr Val Ile Gly Asn Trp

180 185 190

His Leu Arg Gly Val Ser Gly Gly Glu Lys Arg Arg Val Ser Ile Ala

195 200 205

Leu Glu Ile Leu Met Arg Pro Arg Leu Leu Phe Leu Asp Glu Pro Thr

210 215 220

Ser Gly Leu Asp Ser Ala Ser Ala Phe Phe Val Thr Gln Thr Leu Arg

225 230 235 240

Gly Leu Ala Arg Asp Gly Arg Thr Val Ile Ala Ser Ile His Gln Pro

245 250 255

Ser Ser Glu Val Phe Gln Leu Phe Asp Arg Leu Tyr Leu Leu Ser Gly

260 265 270

Gly Lys Thr Val Tyr Phe Gly Gln Ala Ser Glu Ala Cys Glu Phe Phe

275 280 285

Ala Gln Ala Gly Phe Pro Cys Pro Ala Leu Arg Asn Pro Ser Asp His

290 295 300

Phe Leu Arg Cys Ile Asn Ala Asp Phe Asp Lys Val Lys Ala Thr Leu

305 310 315 320

Lys Gly Ser Met Lys Met Arg Phe Glu Arg Ser Asp Asp Pro Leu Glu

325 330 335

Arg Ile Thr Thr Cys Glu Ala Ile Arg Lys Leu Phe Ser His Tyr Gln

340 345 350

His Ser Gln Asp Tyr Leu Ala Thr Arg Gln Lys Val Asp Glu Met Ala

355 360 365

Arg Val Lys Gly Thr Val Leu Asp Ala Gly Gly Ser Gln Ala Ser Phe

370 375 380

Gly Met Gln Ala Cys Thr Leu Thr Lys Arg Ser Phe Val Asn Met Thr

385 390 395 400

Arg Asp Phe Gly Tyr Tyr Trp Leu Arg Leu Val Ile Tyr Val Val Val

405 410 415

Thr Val Cys Ile Gly Ser Ile Tyr Leu Asn Val Gly Thr Lys Tyr Ser

420 425 430

Ser Ile Leu Ala Arg Gly Ala Cys Ala Ser Phe Ile Phe Gly Phe Val

435 440 445

Thr Phe Met Ser Ile Gly Gly Phe Pro Ser Phe Val Glu Asp Met Lys

450 455 460

Val Phe Gln Arg Glu Arg Leu Asn Gly His Tyr Gly Val Ala Ala Phe

465 470 475 480

Val Leu Gly Asn Thr Ala Ser Ala Ala Pro Phe Leu Leu Leu Ile Thr

485 490 495

Val Cys Ser Gly Thr Leu Cys Tyr Phe Met Val Gly Leu His Pro Gly

500 505 510

Leu Ser His Tyr Val Phe Phe Leu Leu Cys Leu Tyr Ala Ser Val Thr

515 520 525

Val Val Glu Ser Leu Met Met Ala Ile Ala Ser Val Val Pro Asn Phe

530 535 540

Leu Met Gly Ile Ile Thr Gly Ala Gly Ile Gln Gly Val Phe Met Leu

545 550 555 560

Val Ser Gly Tyr Phe Arg Leu Pro His Asp Ile Pro Lys Pro Phe Trp

565 570 575

Arg Tyr Pro Met Ser Tyr Ile Ser Phe His Tyr Trp Ala Leu Gln Gly

580 585 590

Gln Tyr Gln Asn Asp Leu Val Gly Leu Met Phe Asp Asn Gln Ser Glu

595 600 605

Leu Leu Pro Lys Ile Pro Gly Glu Tyr Val Leu Glu Asn Val Phe Gln

610 615 620

Ile Asp Val Gly Arg Ser Lys Trp Leu Asp Leu Ser Val Leu Phe Ala

625 630 635 640

Met Ile Val Val Tyr Arg Leu Leu Phe Phe Ala Met Ile Lys Val Ser

645 650 655

Glu Asp Val Thr Pro Trp Leu Arg Arg Tyr Ile Ala Arg Arg Arg Val

660 665 670

Gln Arg Arg Arg Gly Arg Ala Arg Asp Ala Ala Ala Glu Leu Gln Val

675 680 685

Gly Arg Ser Pro Ser Leu Arg Gly Tyr Val Val Asp Asp Asp Leu Pro

690 695 700

Ala Asp His Pro

705

<210> 24

<211> 25

<212>DNA

<213> Synthetic

<400> 24

ggctgatcac ctactacaag aactc 25

<210> 25

<211> 20

<212>DNA

<213> Synthetic

<400> 25

ctcctgcatc cagcactgtc 20

<210> 26

<211> 20

<212>DNA

<213> Synthetic

<400> 26

gctatgtacg tcgccatcca 20

<210> 27

<211> 21

<212>DNA

<213> Synthetic

<400> 27

ggacagtgtg gctgacacca t 21

<210> 28

<211> 27

<212>DNA

<213> Synthetic

<400> 28

actggtaccc ttaatccctt ctatacc 27

<210> 29

<211> 28

<212>DNA

<213> Synthetic

<400> 29

tctgtcgact catctctccc tctctcgc 28

<210> 30

<211> 27

<212>DNA

<213> Synthetic

<400> 30

actggtaccc ttaatccctt ctatacc 27

<210> 31

<211> 31

<212>DNA

<213> Synthetic

<400> 31

agtggatccg gatcatggaa atatcaagta c 31

<210> 32

<211> 29

<212>DNA

<213> Synthetic

<400> 32

ctcagatctg atccaagtgg ctggacctc 29

<210> 33

<211> 31

<212>DNA

<213> Synthetic

<400> 33

ctcactagtc agctgctgat atacaccttg a 31

<210> 34

<211> 244

<212>DNA

<213> Rice (Oryza sativa Japonica)

<400> 34

agagagaggc tcaatgggca ctatggcgtg ctggcattcg tgataagcaa cacgatctcg 60

gcaatgccat tcttgatctt gatcaccttc atctccggga caatgtgcta cttcatggtg 120

cgcctccacc cgggcttcac gcactacctc ttcttcgtcc tctgcctcta cgcaagcgtc 180

accgtcgtcg agagcttgat gatggccatt gccagtgtca tccccaactt cctcatgggc 240

atca 244

<210> 35

<211> 244

<212>DNA

<213> Rice (Oryza sativa Japonica)

<400> 35

tgatgcccat gaggaagttg gggatgacac tggcaatggc catcatcaag ctctcgacga 60

cggtgacgct tgcgtagagg cagaggacga agaagaggta gtgcgtgaag cccgggtgga 120

ggcgcaccat gaagtagcac attgtcccgg agatgaaggt gatcaagatc aagaatggca 180

ttgccgagat cgtgttgctt atcacgaatg ccagcacgcc atagtgccca ttgagcctct 240

ctct 244

<210> 36

<211> 29

<212>DNA

<213> Synthetic

<400> 36

ctcagatctg atccaagtgg ctggacctc 29

<210> 37

<211> 31

<212>DNA

<213> Synthetic

<400> 37

ctcactagtc agctgctgat atacaccttg a 31

<210> 38

<211> 480

<212>DNA

<213> Rice (Oryza sativa Japonica)

<400> 38

gatccaagtg gctggacctc gccgtgctct tcagcatgat cttcatctac cgcctcctct 60

tcttcgccat gatcaaggtc agcgaggacg tcaccccgtg ggtgcgcggc tacgtcgcgc 120

ggcgccgggt gcagggcaag ggcgcccgcg gccgcggggc ggacctctcc gccgcgcgct 180

cgccgtcgct ccgcgcctac gtcgtcgacg ccgccgacga cctgccgccg gcctgaccac 240

ccgtgagaca gacacggctt ctgaaaactc tgaaactctg aactcctcaa ctgaacaagt 300

ggagacagag gaagaaaaag agagagggg agaggctaaa actgaactaaaaaaaaaaga 360

acctgaatat ctgatatgat aattgtgtgttatagtagta gcaatgctta attaatcgat 420

caagctgtgttgttgtattg tattgataat tactaatatc aaggtgtata tcagcagctg 480

<210> 39

<211> 480

<212>DNA

<213> Rice (Oryza sativa Japonica)

<400> 39

cagctgctga tatacacctt gatattagta attatcaata caatacaaca acacagcttg 60

atcgattaat taagcattgc tactactata acacacaatt atcatatcag atattcaggt 120

tctttttttt ttagttcagt tttagcctct ccctctctct ctttttcttc ctctgtctcc 180

acttgttcag ttgaggagtt cagagtttca gagttttcag aagccgtgtc tgtctcacgg 240

gtggtcaggc cggcggcagg tcgtcggcgg cgtcgacgac gtaggcgcgg agcgacggcg 300

agcgcgcggc ggagaggtcc gccccgcggc cgcgggcgcc cttgccctgc acccggcgcc 360

gcgcgacgta gccgcgcacc cacggggtga cgtcctcgct gaccttgatc atggcgaaga 420

agaggaggcg gtagatgaag atcatgctga agagcacggc gaggtccagc cacttggatc 480

Claims (6)

1. a method for regulating plant fertility, regulates the fertility of plants by enhancing or reducing the expression activity of fertility gene, it is characterized in that, the DNA sequence of described fertility gene is selected from one of the sequences of following group: (a) the nucleotide sequence shown in SEQ ID NO: 1; (b) the amino acid sequence encoded by it is the nucleotide sequence shown in SEQ ID NO: 2; or (c) DNA sequence complementary to the nucleotide sequence described in (a) or (b). 2. An application of an expression cassette in regulating plant fertility, characterized in that the expression cassette comprises one of the nucleotide sequences selected from the following groups: (a) the nucleotide sequence shown in SEQ ID NO: 1; (b) the amino acid sequence encoded by it is the nucleotide sequence shown in SEQ ID NO: 2; or (c) DNA sequence complementary to the nucleotide sequence described in (a) or (b). 3. An application of an expression vector in regulating plant fertility, characterized in that the expression vector comprises one of the nucleotide sequences selected from the following groups: (a) the nucleotide sequence shown in SEQ ID NO: 1; (b) the amino acid sequence encoded by it is the nucleotide sequence shown in SEQ ID NO: 2; or (c) DNA sequence complementary to the nucleotide sequence described in (a) or (b). 4. The application of a mutant material, the mutant material is caused by the mutation of the nucleotide sequence, and the plant containing the nucleotide sequence after the mutation shows male sterility, and it is characterized in that the mutant material can be produced by mutation The nucleotide sequence of the male sterility phenotype is the nucleotide sequence of the rice FG3 gene, and the nucleotide sequence produced by the mutation is shown in SEQ ID NO:7. 5. The application of claim 4, wherein the nucleotide sequence of the rice FG3 gene is selected from one of the sequences of the following groups: (a) have the nucleotide sequence shown in SEQ ID NO: 1; (b) the amino acid sequence encoded by it is the nucleotide sequence shown in SEQ ID NO: 2; or (c) a DNA sequence complementary to the sequence described in (a) or (b). 6. The application according to any one of claims 4 or 5, wherein the mutation can be point mutation, DNA deletion or insertion mutation, or gene silencing means such as RNAi and site-directed mutation.