Use of irx10 protein in regulating development of plant vessel and crop growth

Abstract

The application discloses application of IRX10 protein in regulation of plant vessel development and crop growth. The IRX10 protein can be a protein with a coding sequence being sequence 1 in a sequence table. Experiments prove that the IRX10 protein has the function of regulating plant vessel development and crop growth, can regulate specific deposition of xylan around vessel pit by changing xylan content, forms nanoscale domains, and thus influences the structure and function of the plant vessel. The application has great value for cultivating transgenic plants with optimized vessel function, and plays a significant role in improving crop growth and yield, and provides economic value.

Description

Technical Field

[0001] This invention belongs to the field of genetic engineering, specifically involving the application of IRX10 protein in regulating plant vascular development and crop growth. Background Technology

[0002] Plant cell walls are unique structures in plant cells, providing vital functions such as mechanical support and water transport. Xylan is an important polysaccharide component of plant cell walls, cross-linking with polymers such as cellulose and lignin to form a complex network structure that ultimately determines the structure and function of the cell wall. Defects in xylan synthesis can lead to phenotypes such as reduced plant support, slow growth and development, and even death.

[0003] Vascular tissue, distributed throughout various plant organs, is a vital organ for transporting water and nutrients, primarily composed of xylem and phloem. The regionalized distribution of polysaccharides can regulate the differentiation of xylem vessels, forming protoxylem with spiral or annular patterns, or metaxylem with reticulate or pitted patterns. This differentiation allows for precise regulation of xylem vessel function, such as water transport, prevention of air embolism, and inhibition of pathogen invasion. How plants regulate the regionalization of cell wall polysaccharides remains to be studied. Therefore, elucidating the mechanisms of plant cell wall polysaccharide regionalization can provide guidance for regulating vessel development and offer targets for molecular design improvements in cultivating high-quality, high-yield crops. Summary of the Invention

[0004] The purpose of this invention is to provide the application of the IRX10 protein in regulating plant vascular development and / or crop growth.

[0005] To achieve the above objectives, the present invention provides the use of a protein or substance regulating gene expression in regulating plant height and / or plant vascular development, or in the preparation of products regulating plant height and / or plant vascular development, wherein the gene encodes the protein, the protein being P1 or P2, wherein P1 is IRX10-IRX10L-IRX10L3, IRX10-IRX10L4-IRX10L5, or IRX10L-IRX10L2-IRX10L3, wherein IRX10-IRX10L-IRX10L3 is composed of IRX10, IRX10L, and IRX10L3, wherein IRX10-IRX10L4-IRX10L5 is composed of IRX10, IRX10L4, and IRX10L5, and wherein IRX10L-IRX10L2-IRX10L3 is composed of IRX10L, IRX10L2, and IRX10L3.

[0006] The P2 is selected from one or two of the IRX10, IRX10L, IRX10L3, RX10L4, and IRX10L5.

[0007] The IRX10 is any one of the following A1)-A3):

[0008] A1) Proteins encoded by DNA molecules whose coding sequence is shown in SEQ ID No. 1;

[0009] A2) A protein obtained by substituting and / or deleting and / or adding one or more amino acid residues from the amino acid sequence shown in A1), which has more than 80% identity with the protein shown in A1) and is related to plant height and / or plant vascular development.

[0010] A3) A fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of A1) or A2);

[0011] The IRX10L is any one of the following (A11)-A13):

[0012] A11) A protein encoded by a DNA molecule whose coding sequence is shown in SEQ ID No. 3;

[0013] A12) A protein obtained by substituting and / or deleting and / or adding one or more amino acid residues from the amino acid sequence shown in A11) and having more than 80% identity with the protein shown in A11) and being related to plant height and / or plant vascular development.

[0014] A13) is a fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of A11) or A12);

[0015] The IRX10L2 is any one of the following (A21)-A23):

[0016] A21) A protein encoded by a DNA molecule whose coding sequence is shown in SEQ ID No. 5;

[0017] A22) A protein obtained by substituting and / or deleting and / or adding one or more amino acid residues from the amino acid sequence shown in A21), which has more than 80% identity with the protein shown in A21) and is related to plant height and / or plant vascular development.

[0018] A23) is a fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of A21) or A22);

[0019] The IRX10L3 is any one of the following (A31)-A33):

[0020] A31) A protein encoded by a DNA molecule whose coding sequence is shown in SEQ ID No. 7;

[0021] A32) A protein obtained by substituting and / or deleting and / or adding one or more amino acid residues from the amino acid sequence shown in A31), which has more than 80% identity with the protein shown in A31) and is related to plant height and / or plant vascular development.

[0022] A33) is a fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of A31) or A32);

[0023] The IRX10L4 is any one of the following (A41)-A43):

[0024] A41) A protein encoded by a DNA molecule whose coding sequence is shown in SEQ ID No. 9;

[0025] A42) A protein obtained by substituting and / or deleting and / or adding one or more amino acid residues from the amino acid sequence shown in A41), which has more than 80% identity with the protein shown in A41) and is related to plant height and / or plant vascular development.

[0026] A43) is a fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of A41) or A42);

[0027] The IRX10L5 is any one of the following (A51)-A53):

[0028] A51) A protein encoded by a DNA molecule whose coding sequence is shown in SEQ ID No. 11;

[0029] A52) A protein obtained by substituting and / or deleting and / or adding one or more amino acid residues from the amino acid sequence shown in A51), which has more than 80% identity with the protein shown in A51 and is related to plant height and / or plant vascular development.

[0030] The fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of A53) or A51) or A52) is a protein.

[0031] In the above applications, the substance regulating gene expression can be a substance that performs at least one of the following six types of regulation: 1) regulation at the gene transcription level; 2) post-transcriptional regulation of the gene (i.e., regulation of splicing or processing of the primary transcript of the gene); 3) regulation of RNA transport of the gene (i.e., regulation of mRNA transport of the gene from the nucleus to the cytoplasm); 4) regulation of gene translation; 5) regulation of mRNA degradation of the gene; and 6) post-translational regulation of the gene (i.e., regulation of the activity of the protein translated from the gene).

[0032] In the above applications, the regulation of gene expression can be achieved by inhibiting or reducing gene expression, which can be achieved by gene knockout or gene silencing.

[0033] Gene knockout refers to the phenomenon of inactivating a specific target gene through homologous recombination. Gene knockout inactivates a specific target gene by altering its DNA sequence.

[0034] Gene silencing refers to the phenomenon of preventing or reducing gene expression without damaging the original DNA. Gene silencing presupposes no change in the DNA sequence, resulting in the absence or reduction of gene expression. Gene silencing can occur at two levels: transcriptional silencing due to DNA methylation, heterochromatinization, and position effects; and post-transcriptional gene silencing, which inactivates the gene at the post-transcriptional level through specific inhibition of target RNA. This includes antisense RNA, co-suppression, gene quelling, RNA interference (RNAi), and microRNA (miRNA)-mediated translational repression.

[0035] In the above applications, the substance regulating gene expression can be a reagent that inhibits or reduces the expression of the gene. The reagent that inhibits or reduces gene expression can be a gene knockout reagent, such as a reagent that knocks out the gene through homologous recombination or a reagent that knocks out the gene through CRISPR-Cas9. The reagent that inhibits or reduces gene expression can contain nucleotides targeting the gene, such as siRNA, shRNA, sgRNA, miRNA, or antisense RNA, or can be a vector expressing the nucleotides.

[0036] Furthermore, in the above applications, the substance regulating the expression of the protein-coding gene is a biological material, and the biological material is any one of the following:

[0037] B1) Nucleic acid molecules that encode the above proteins;

[0038] B2) An expression cassette containing the nucleic acid molecule described in B1);

[0039] B3) A recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B2);

[0040] B4) Recombinant microorganisms containing the nucleic acid molecules described in B1), or recombinant microorganisms containing the expression cassette described in B2), or recombinant microorganisms containing the recombinant vector described in B3);

[0041] B5) A transgenic plant cell line containing the nucleic acid molecule described in B1), or a transgenic plant cell line containing the expression cassette described in B2), or a transgenic plant cell line containing the recombinant vector described in B3);

[0042] B6) Transgenic plant tissue containing the nucleic acid molecules described in B1), or transgenic plant tissue containing the expression cassette described in B2), or transgenic plant tissue containing the recombinant vector described in B3);

[0043] B7) A transgenic plant organ containing the nucleic acid molecule described in B1), or a transgenic plant organ containing the expression cassette described in B2), or a transgenic plant organ containing the recombinant vector described in B3);

[0044] B8) Nucleic acid molecules that inhibit or reduce the expression of the genes encoding the above proteins or the activity of the above proteins;

[0045] B9) Expression cassettes, recombinant vectors, recombinant microorganisms, or transgenic plant cell lines containing the nucleic acid molecules described in B8).

[0046] Furthermore, in the above applications, the nucleic acid molecule described in B8) is either a DNA molecule expressing a gRNA that targets the protein-coding gene or a gRNA that targets the protein-coding gene.

[0047] Furthermore, in the above applications, the target sequence of the sgRNA includes:

[0048] C1) Nucleotides 1918-1937 of sequence 2 in the sequence listing (i.e., 5′-GCCACCCTGGTTCAGACCTT-3′), nucleotides 1325-1344 of sequence 4 in the sequence listing (i.e., 5′-TGACCCTGTCGGGAGGCTTA-3′), and nucleotides 444-463 of sequence 8 in the sequence listing (i.e., 5′-CGACGACGCAGAAGCATGGC-3′);

[0049] C2) Nucleotides 242-260 of sequence 2 in the sequence listing (i.e., 5′-AAGATGAGGAGGTGGGTCT-3′), nucleotides 153-172 of sequence 10 in the sequence listing (i.e., 5′-GGCGCTCATGGTGGGCGCAA-3′), and nucleotides 242-260 of sequence 12 in the sequence listing (i.e., 5′-TCGGTGATGGGATCAAGAA-3′);

[0050] Nucleotides 1325-1344 of sequence 4 in the C3 sequence listing (i.e., 5′-TGACCCTGTCGGGAGGCTTA-3′), nucleotides 1066-1084 of sequence 6 in the sequence listing (i.e., 5′-CTAGAGAAGACGATCCTGT-3′), and nucleotides 444-463 of sequence 8 in the sequence listing (i.e., 5′-CGACGACGCAGAAGCATGGC-3′).

[0051] In further applications described above, the regulation of plant height and / or plant vascular development can be achieved by reducing plant height and / or inhibiting plant vascular development.

[0052] Furthermore, the recipient plant mentioned in the above applications can be any one of E1)-E5):

[0053] E1) Monocotyledons;

[0054] E2) Plants of the order Poales;

[0055] E3) Gramineae plants;

[0056] E4) Plants of the genus *Oryza*;

[0057] E5) Rice.

[0058] To achieve the above objective, in a second aspect, the present invention provides a method for reducing rice plant height, comprising knocking out the gene encoding the aforementioned protein in the rice genome to obtain rice with reduced plant height.

[0059] Furthermore, in the above method, the gene encoding the protein in the rice genome that is knocked out can be any one of the following 1)-3): 1) knocking out the gene encoding IRX10, the gene encoding IRX10L, and the gene encoding IRX10L3 in the rice genome; 2) knocking out the gene encoding IRX10, the gene encoding IRX10L4, and the gene encoding IRX10L5 in the rice genome; 3) knocking out the gene encoding IRX10L, the gene encoding IRX10L2, and the gene encoding IRX10L3 in the rice genome.

[0060] Further, in the above method, step 1) can be inserting nucleotide A between nucleotides 1934 and 1935 in sequence 2 of the rice genome sequence listing, that is, inserting a single base A between nucleotides 560-561 in the IRX10 gene coding sequence shown in sequence 1 of the sequence listing; inserting nucleotide C between nucleotides 1340 and 1341 in sequence 4 of the rice genome sequence listing, that is, inserting a single base C between nucleotides 144-145 in the IRX10L gene coding sequence shown in sequence 3 of the sequence listing; and inserting nucleotide T between nucleotides 459 and 460 in sequence 8 of the rice genome sequence listing, that is, inserting a single base T between nucleotides 110-111 in the IRX10L3 gene coding sequence shown in sequence 7 of the sequence listing;

[0061] 2) can be an insertion of nucleotide T between nucleotides 257 and 258 of sequence 2 in the rice genome sequence listing, i.e., an insertion of a single base T between nucleotides 13-14 of the IRX10 gene coding sequence shown in sequence 1 of the sequence listing; an insertion of nucleotide T between nucleotides 169 and 170 of sequence 10 in the rice genome sequence listing, i.e., an insertion of a single base T between nucleotides 10-11 of the IRX10L4 gene coding sequence shown in sequence 9 of the sequence listing; and an insertion of nucleotide A between nucleotides 257 and 258 of sequence 12 in the rice genome sequence listing, i.e., an insertion of a single base A between nucleotides 10-11 of the IRX10L5 gene coding sequence shown in sequence 11 of the sequence listing.

[0062] 3) can be an insertion of nucleotide C between nucleotides 1341 and 1342 in sequence 4 of the rice genome sequence listing, i.e., an insertion of a single base C between nucleotides 145-146 in the IRX10L gene coding sequence shown in sequence 3 of the sequence listing; a deletion of nucleotide 1081 in sequence 6 of the rice genome sequence listing, i.e., a deletion of nucleotide 140 in the IRX10L2 gene coding sequence shown in sequence 5 of the sequence listing; and an insertion of nucleotide T between nucleotides 460 and 461 in sequence 8 of the rice genome sequence listing, i.e., an insertion of a single base T between nucleotides 111-112 in the IRX10L3 gene coding sequence shown in sequence 7 of the sequence listing.

[0063] The present invention also provides the above-mentioned proteins or substances that regulate gene expression.

[0064] In this invention, the protein shown in A1) consists of 417 amino acid residues; the protein shown in A11) consists of 415 amino acid residues; the protein shown in A21) consists of 417 amino acid residues; the protein shown in A31) consists of 422 amino acid residues; the protein shown in A41) consists of 434 amino acid residues; and the protein shown in A51) consists of 420 amino acid residues.

[0065] The proteins mentioned above can be synthesized artificially, or their encoding genes can be synthesized first and then expressed biologically.

[0066] The protein tag refers to a polypeptide or protein fused with a target protein using in vitro DNA recombination technology for expression, detection, tracing, and / or purification of the target protein. The protein tag may be a Flag protein tag, His protein tag, MBP protein tag, HA protein tag, myc protein tag, GST protein tag, and / or SUMO protein tag, etc.

[0067] In this invention, identity refers to the similarity of amino acid sequences or nucleotide sequences. The identity of amino acid sequences can be determined using homology search sites on the Internet, such as the BLAST page on the NCBI homepage. For example, in Advanced BLAST 2.1, using blastp as the program, setting the Expect value to 10, setting all filters to OFF, using BLOSUM62 as the matrix, setting the Gap existence cost, Per residue gap cost, and Lambdaratio to 11, 1, and 0.85 (default values) respectively, and performing an identity search on a pair of amino acid sequences to calculate the identity value (%), then the identity value can be obtained.

[0068] In this invention, the 80% or more identity can be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity. This invention provides the function of IRX10 and its homologous proteins in regulating plant vascular development and crop growth. In wild-type Nipponbare, this invention altered the expression level of IRX10-related proteins, obtaining transgenic plants with reduced cell wall xylan content. Experiments demonstrated that IRX10 protein has the function of synthesizing plant cell wall xylan, which can be used to regulate the specific deposition of xylan around plant vascular pits, forming nano-lattice structures, thereby affecting the structure and function of plant vascular bundles. This has significant value for cultivating transgenic plants with optimized vascular function, and will play a significant role in improving crop growth and yield, providing economic value. Attached Figure Description

[0069] Figure 1 Diagram showing the construction of the CRISPR / Cas9 recombination vector.

[0070] Figure 2 The images show the phenotype of IRX10 gene-related mutant materials and the xylose content in the stem cell walls. Specifically: A is a phenotypic diagram of the IRX10 gene-related mutant materials, with a scale bar of 12 cm; B is a diagram showing the xylose content measured in the stem cell walls of the corresponding genetic materials.

[0071] Figure 3 The images show the stem development of IRX10 gene-related mutant materials, where: A is a cross-sectional view of the vascular bundles in the stem of the IRX10 gene-related mutant material, with a scale bar of 20 μm; B is a magnified view of the xylem vessels in the material in Figure A, with a scale bar of 10 μm; C is a statistical graph of the vessel area in Figure B; D is a display of the vessel cell wall thickness in the IRX10 gene-related mutant material, with a scale bar of 1 μm; and E is a statistical graph of the vessel cell wall thickness in Figure D.

[0072] Figure 4 The diagram shows the morphology of vessel pits in IRX10 gene-related mutant materials. In the diagram, A is a display of vessel pits in IRX10 gene-related mutant materials, with a scale bar of 2 μm; B is a statistical graph of the vessel pit area corresponding to the diagram in A.

[0073] Figure 5 The images show xylan localization maps of vessel pits in IRX10 gene-related mutant materials, where: A is a xylan localization map of vessel pits in IRX10 gene-related mutant materials, with a scale bar of 2 μm; B is a longitudinal view of map A, with a scale bar of 1 μm.

[0074] Figure 6 To detect the transport capacity of rhodamine B in ducts of IRX10 gene-related mutant materials using a rhodamine B transport assay; where: A is a microscopic image of rhodamine B transport in IRX10 gene-related mutant materials, scale bar is 500 μm; B is a statistical graph of transported rhodamine B content. RB represents rhodamine B solution.

[0075] Figure 7 To determine the leaf transpiration potential in IRX10 gene-related mutant materials. Detailed Implementation

[0076] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0077] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0078] Unless otherwise specified, the quantitative experiments in the following examples are all repeated three times, and the results are averaged.

[0079] The following examples used SPSS 19.0 statistical software to process the data. The experimental results are expressed as mean ± standard deviation. The graph test was used, and different lowercase letters in the results indicate that the differences are significant at the 0.05 level.

[0080] The japonica rice variety “Nipponbare” (WT, also known as wild-type plant) in the following examples was purchased from the China National Rice Research Institute.

[0081] The Agrobacterium EHA105 used in the following examples was purchased from CAMIA, Australia.

[0082] The cloned strain DH5α used in the following examples was purchased from TAKARA Corporation, Japan.

[0083] The expression cassettes pYLsgRNA-OsU3m / LacZ, pYLsgRNA-OsU6a, and the vector pYLCRISPR / Cas9Pubi-H in the following examples were kindly provided by Professor Liu Yaoguang's research group at the State Key Laboratory of Subtropical Agricultural Bioresources Conservation and Utilization, College of Life Sciences, South China Agricultural University. These materials are disclosed in the literature "Dong Z, Liu YG*. 2015. A robust CRISPR / Cas9 system for convenient high-efficiency multiplex genome editing in monocot and dicot plants. Molecular Plant. 8: 1274-1284". The public can obtain the above-mentioned biological materials from the applicant. These biological materials are only for repeating the experiments of this invention and should not be used for other purposes.

[0084] Example 1: Obtaining rice genetic materials related to the IRX10 gene

[0085] 1.1 Obtaining the rice brittle stem mutant

[0086] 1.1.1 Phenotype of rice brittle stem mutant

[0087] The rice brittle culm 18 (abbreviated as bc18) is a material obtained by spontaneous mutation of the japonica rice variety "Nipponbare". Compared with the wild type "Nipponbare", the main phenotypic characteristics of the mutant bc18 are: (1) the stem becomes brittle (the mechanical strength of the stem is significantly reduced and the xylem vessel structure in the stem is abnormal); (2) the plant height is shorter.

[0088] The phenotypes of the above genetic materials are shown in the figure. Figure 2 A.

[0089] 1.1.2 IRX10 protein sequence analysis of rice brittle stem mutant

[0090] Map-based cloning identified the target gene as IRX10 and its point mutation location. The gene encoding the IRX10 protein is the DNA molecule shown in SEQ ID No. 1, and the genomic sequence of IRX10 is the DNA molecule shown in SEQ ID No. 2.

[0091] Sequencing revealed that, compared to the wild-type rice plant (Nipponbare), the bc18 IRX10 genomic sequence showed a G-to-A mutation at position 2103. This mutation resulted in an arginine-to-lysine change at position 295 of the IRX10 protein sequence; the remaining amino acid sequences were identical to those of the wild-type rice plant.

[0092] 1.2 Obtaining IRX10 gene-related mutants

[0093] 1.2.1. Construction of CRISPR-Cas9 gene knockout vector

[0094] In rice, the IRX10 gene has five homologous sequences: IRX10L, IRX10L2, IRX10L3, IRX10L4, and IRX10L5. Specifically, the gene encoding the IRX10L protein is the DNA molecule shown in SEQ ID No. 3, and the genomic sequence of IRX10L is the DNA molecule shown in SEQ ID No. 4; the gene encoding the IRX10L2 protein is the DNA molecule shown in SEQ ID No. 5, and the genomic sequence of IRX10L2 is the DNA molecule shown in SEQ ID No. 6; the gene encoding the IRX10L3 protein is the DNA molecule shown in SEQ ID No. 7, and the genomic sequence of IRX10L3 is the DNA molecule shown in SEQ ID No. 8; the gene encoding the IRX10L4 protein is the DNA molecule shown in SEQ ID No. 9, and the genomic sequence of IRX10L4 is the DNA molecule shown in SEQ ID No. 10; and the gene encoding the IRX10L5 protein is the DNA molecule shown in SEQ ID No. 11, and the genomic sequence of IRX10L5 is the DNA molecule shown in SEQ ID No. 12.

[0095] Step 1) Identify suitable nucleotide sequences 20 bp upstream of NGG in IRX10 and its homologous gene sequences as target sequences for gRNA, excluding non-specific target cleavage sites. The target sequences for each gene are as follows:

[0096] IRX10 target sequence 1: 5'-GCCACCCTGGTTCAGACCTT-3' (positions 1918-1937 of sequence 2 in the sequence listing);

[0097] IRX10 target sequence 2: 5'-AAGATGAGGAGGTGGGTCT-3' (positions 242-260 of sequence 2 in the sequence listing);

[0098] IRX10L target sequence: 5'-TGACCCTGTCGGGAGGCTTA-3' (positions 1325-1344 of sequence 4 in the sequence listing);

[0099] IRX10L2 target sequence: 5'-CTAGAGAAGACGATCCTGT-3' (positions 1066-1084 of sequence 6 in the sequence listing);

[0100] IRX10L3 target sequence: 5'-CGACGACGCAGAAGCATGGC-3' (positions 444-463 of sequence 8 in the sequence listing);

[0101] IRX10L4 target sequence: 5'-GGCGCTCATGGTGGGCGCAA-3' (positions 153-172 of sequence 10 in the sequence listing);

[0102] IRX10L5 target sequence: 5'-TCGGTGATGGGATCAAGAA-3' (positions 242-260 of sequence 12 in the sequence listing);

[0103] Primers were designed based on the target sequence, as follows:

[0104] IRX10-target-F1: 5'-GGCAGCCACCCTGGTTCAGACCTT-3';

[0105] IRX10-target-R1: 5'-AAACAAGGTCTGAACCAGGGTGGC-3';

[0106] IRX10-target-F2: 5'-GGCAAAGATGAGGAGGTGGGTCT-3';

[0107] IRX10-target-R2: 5'-AAACAGACCCACCTCCTCATCTT-3';

[0108] IRX10L-target-F: 5'-GCCGTGACCCTGTCGGGAGGCTTA-3';

[0109] IRX10L-target-R: 5'-AAACTAAGCCTCCCGACAGGGTCA-3';

[0110] IRX10L2-target-F: 5'-GCCGCTAGAGAAGACGATCCTGT-3';

[0111] IRX10L2-target-R: 5'-AAACACAGGATCGTCTTCTCTAG-3';

[0112] IRX10L3-target-F: 5'-GCCGCGACGACGCAGAAGCATGGC-3';

[0113] IRX10L3-target-R: 5'-AAACGCCATGCTTCTGCGTCGTCG-3';

[0114] IRX10L4-target-F: 5'-GCCGGGCGCTCATGGTGGGCGCAA-3';

[0115] IRX10L4-target-R: 5'-AAACTTGCGCCCACCATGAGCGCC-3';

[0116] IRX10L5-target-F: 5'-GCCGTCGGGTGATGGGATCAAGAA-3';

[0117] IRX10L5-target-R: 5'-AAACTTCTTGATCCCATCACCGA-3'.

[0118] Step 2) Gene amplification is performed using primer pairs. The gene expressing the sgRNA targeting the target sequence in Step 1) is ligated to the pYLgRNA-OsU3m / LacZ (represented by U3pr in the map) and pYLgRNA-OsU6a (represented by U6a pr in the map) gene expression cassettes, and the above expression cassettes are then linked together. The combination method and ligation order are described in [reference needed]. Figure 1 ;

[0119] Step 3) Connect the expression cassettes from step 2) according to the designed combination and link them to the binary vector pYLCRISPR / Cas9Pubi-H (the original name of the vector was pYLCRISPR / Cas9-MH, hereinafter referred to as pYLCRISPR / Cas9-MH). This will yield plasmids MH-IRX10-IRX10L-IRX10L3 (expression vector for three gRNAs: gRNA targeting IRX10 target sequence 1, gRNA targeting IRX10L target sequence, and gRNA targeting IRX10L3 target sequence) and MH-IRX10-IRX10L4-IRX10L5 (expression vector for gRNAs targeting IRX10L1 and IRX10L3 target sequence). Expression vectors for three gRNAs: target sequence 2 gRNA, gRNA targeting IRX10L4 gRNA, and gRNA targeting IRX10L5 gRNA; and MH-IRX10L-IRX10L2-IRX10L3 (expression vectors for three gRNAs: gRNA targeting IRX10L, gRNA targeting IRX10L2, and gRNA targeting IRX10L3); The maps of plasmids MH-IRX10-IRX10L-IRX10L3, MH-IRX10-IRX10L4-IRX10L5, and MH-IRX10L-IRX10L2-IRX10L3 are shown below. Figure 1 As shown.

[0120] Step 4) Transform the plasmid from step 3) into E. coli DH5α competent cells, pick white plaques for culture, extract plasmids for sequencing, and retain plasmids that are correctly sequenced.

[0121] 1.2.2. Obtaining transgenic plants

[0122] Step 1) Culture of rice embryos

[0123] Wild-type rice Nipponbare seeds were dehulled and sterilized as follows: 70% ethanol for 3 minutes, 0.1% mercuric chloride solution for 5 minutes, 10% sodium hypochlorite solution for 20 minutes, and rinsed 3-4 times with sterile distilled water. The sterilized seeds were then spread on NB medium. Callus tissue grew from the mature embryo scutellum after 20 days and was subcultured onto NB medium, with subsequent subcultures every two weeks.

[0124] Step 2) Preparation of recombinant straw fungal suspension

[0125] The recombinant plasmids MH-IRX10-IRX10L-IRX10L3, MH-IRX10-IRX10L4-IRX10L5, and MH-IRX10L-IRX10L2-IRX10L3 described in section 1 were individually introduced into Agrobacterium tumefaciens EHA105 via electroporation. The cells were cultured in YEP liquid medium until the bacterial concentration at OD600nm was 0.8-1.0, yielding recombinant Agrobacterium tumefaciens EHA105 / MH-IRX10-IRX10L-IRX10L3, EHA105 / MH-IRX10-IRX10L4-IRX10L5, and EHA105 / MH-IRX10L-IRX10L2-IRX10L3, respectively.

[0126] Step 3) Co-culture of rice materials and Agrobacterium tumefaciens

[0127] The recombinant Agrobacterium tumefaciens suspension prepared in step 2) was used to separately infect the embryonic callus tissue in step 1), and the callus tissue was shaken and soaked at room temperature for 20 minutes. The callus tissue was then transferred to NB solid medium containing 20 μM acetylsyleugenol and lined with a layer of sterile filter paper, and cultured in the dark at 26°C for 2-3 days.

[0128] Step 4) Screening of resistant callus and plant regeneration

[0129] The callus tissue from step 3) was spread on a selective medium containing 50 mg / L hygromycin and cultured for two weeks. The screening was repeated once. The vigorous resistant callus tissue was transferred to a differentiation medium and allowed to grow into seedlings. The seedlings were then transferred to 1 / 2 MS solid medium to root and grow strong. They were then moved to a greenhouse or field to become T0 generation plants.

[0130] Transgenic plants transformed with plasmid MH-IRX10-IRX10L-IRX10L3 were named mutant Tm1, transgenic plants transformed with plasmid MH-IRX10-IRX10L4-IRX10L5 were named mutant Tm2, ​​and transgenic plants transformed with plasmid MH-IRX10L-IRX10L2-IRX10L3 were named mutant Tm3.

[0131] 1.2.3. Identification of Transgenic Materials

[0132] Step 1) Design primers approximately 300 bp upstream and downstream of the target site in Step 1) of section 1.2.1, as follows:

[0133] IRX10-CAS9-F1: 5'-GCAACTCTGATTTAATTGGAACA-3';

[0134] IRX10-CAS9-R1: 5'-ATGTATTGGAGATAAGCGGAAAA-3';

[0135] IRX10-CAS9-F2: 5'-CCCCTCACCTTCTCAACTCACCA-3';

[0136] IRX10-CAS9-R2: 5'-TAAACAACCTCCATTTGCGTCTG-3';

[0137] IRX10L-CAS9-F: 5'-ACTCTGGTTGGCAAGCTACTCCT-3';

[0138] IRX10L-CAS9-R: 5'-CAATAGGGCCAATGCGAGGAAAT-3';

[0139] IRX10L2-CAS9-F: 5'-AATGGAGGGTATTCTAGTCTTTTC-3';

[0140] IRX10L2-CAS9-R: 5'-TGAACTTGATCGCACTTCTCATC-3';

[0141] IRX10L3-CAS9-F: 5'-GAGCAGAGCTAATCAAAAGCCCTCT-3';

[0142] IRX10L3-CAS9-R: 5'-GCGAATGCAGGATGAGCAATAGA-3';

[0143] IRX10L4-CAS9-F: 5'-GCTGCCTCCTTTGCTTCCTCCT-3';

[0144] IRX10L4-CAS9-R: 5'-CATATCAGTGCGAATGTGCGATT-3';

[0145] IRX10L5-CAS9-F: 5'-CAACCAAATCTTCTTGGCTTCAT-3';

[0146] IRX10L5-CAS9-R: 5'-AAGGAATACAATGGATGGCACAA-3'.

[0147] Step 2) Genomic DNA was extracted from the transgenic materials Tm1, Tm2, ​​and Tm3 obtained in step 1.2.2, and PCR amplification was performed using the primers designed in step 1), yielding a product of approximately 600 bp. The product was reacted with T7 endonuclease at 42°C for 1 hour, followed by DNA electrophoresis. Product verification will result in two outcomes: if the PCR product band can be cleaved by T7 enzyme into two smaller fragments, the corresponding plant is a heterozygous mutant; if the product cannot be cleaved by T7 enzyme, the corresponding plant genotype is homozygous or wild-type. Further, the uncleavable PCR product DNA was mixed with equal amounts of wild-type plant DNA, and PCR amplification was performed using the appropriate primers, followed by T7 enzyme digestion. Product verification will again result in two outcomes: if the PCR product can be cleaved, the corresponding plant is a homozygous mutant; if the PCR product cannot be cleaved, the corresponding plant is wild-type. Finally, the mutation site was determined by sequencing.

[0148] Based on the above methods, homozygous mutants of IRX10, Tm1, Tm2, ​​and Tm3, were obtained. Compared with wild-type rice Nipponbare, the IRX10 homozygous mutant Tm1 showed mutations in the genes of IRX10, IRX10L, and IRX10L3 in the rice genome. In the two homologous chromosomes, the genomic gene of IRX10 (Sequence 2 in the sequence listing) underwent the following changes: In the Nipponbare rice genome, nucleotide A was inserted between nucleotides 1934 and 1935 of Sequence 2 in the sequence listing, i.e., a single base A was inserted between nucleotides 560-561 of the IRX10 gene coding sequence shown in Sequence 1 in the sequence listing, resulting in a frameshift mutation. The translation of the IRX10 protein terminated prematurely after 187 amino acids, thus knocking out the IRX10 gene. In the two homologous chromosomes, the genomic gene of IRX10L (Sequence 4 in the sequence listing) underwent the following changes: In the Nipponbare rice genome, the sequence listing... In sequence 4, a nucleotide C was inserted between nucleotides 1340 and 1341, which is equivalent to a single C insertion between nucleotides 144-145 of the IRX10L gene coding sequence shown in sequence 3 of the sequence listing. This resulted in a frameshift mutation, causing premature termination of IRX10L protein translation after 48 amino acids, thus knocking out the IRX10L gene. In the genomes of two homologous chromosomes, the nucleotide sequence of the IRX10L3 gene in sequence 8 of the sequence listing underwent the following changes: In the genome of rice Nipponbare, a nucleotide T was inserted between nucleotides 459 and 460 of sequence 8 of the sequence listing, which is equivalent to a single T insertion between nucleotides 110-111 of the IRX10L3 gene coding sequence shown in sequence 7 of the sequence listing. This resulted in a frameshift mutation, causing premature termination of IRX10L3 protein translation after 38 amino acids, thus knocking out the IRX10L3 gene.

[0149] Compared to wild-type Nipponbare rice, the IRX10 polygenic homozygous mutant Tm2 exhibits mutations in the genes IRX10, IRX10L4, and IRX10L5 in the rice genome. In the two homologous chromosomes, the genomic gene of IRX10 (Sequence 2 in the sequence listing) shows the following changes: In the Nipponbare rice genome, a nucleotide T is inserted between nucleotides 257 and 258 of Sequence 2 in the sequence listing, specifically between nucleotides 13-14 of the IRX10 gene coding sequence shown in Sequence 1 in the sequence listing. This results in a frameshift mutation, causing premature termination of IRX10 protein translation after 6 amino acids, thus knocking out the IRX10 gene. In the two homologous chromosomes, the genomic gene of IRX10L4 (Sequence 10 in the sequence listing) shows the following changes: In the Nipponbare rice genome, the nucleotide sequence... A nucleotide T was inserted between nucleotides 169 and 170 of sequence 10, i.e., a single nucleotide T was inserted between nucleotides 10 and 11 of the coding sequence of the IRX10L4 gene shown in sequence 9 of the sequence listing. This resulted in a frameshift mutation, causing the translation of the IRX10L4 protein to terminate prematurely after 3 amino acids, thus knocking out the IRX10L4 gene. In the genomes of two homologous chromosomes, the nucleotide sequence of the IRX10L5 gene in sequence 12 of the sequence listing underwent the following changes: In the genome of rice Nipponbare, a nucleotide A was inserted between nucleotides 257 and 258 of sequence 12 of the sequence listing. This resulted in a single nucleotide A inserted between nucleotides 10 and 11 of the coding sequence of the IRX10L5 gene shown in sequence 11 of the sequence listing. This resulted in a frameshift mutation, causing the translation of the IRX10L5 protein to terminate prematurely after 2 amino acids, thus knocking out the IRX10L5 gene.

[0150] In the IRX10 polygenic homozygous mutant Tm3, compared with wild-type rice Nipponbare, mutations were found in the genes IRX10L, IRX10L2, and IRX10L3 in the rice genome. On two homologous chromosomes, the genomic gene of IRX10L (nucleotide sequence 4 in the sequence listing) underwent the following changes: In the Nipponbare rice genome, a nucleotide C was inserted between nucleotides 1341 and 1342 of sequence 4 in the sequence listing, i.e., a single base C was inserted between nucleotides 145-146 of the IRX10L gene coding sequence shown in sequence 3 in the sequence listing, resulting in a frameshift mutation. Translation of the IRX10L protein terminated prematurely after 48 amino acids, thus knocking out the IRX10L gene. On two homologous chromosomes, the genomic gene of IRX10L2 (nucleotide sequence 6 in the sequence listing) underwent the following changes: In the genome of Nipponbare rice, the deletion of nucleotide 1081 in sequence 6 of the sequence listing, which corresponds to the deletion of nucleotide 140 in the coding sequence of the IRX10L2 gene shown in sequence 5 of the sequence listing, results in a frameshift mutation. The translation of the IRX10L2 protein terminates prematurely after 47 amino acids, thus knocking out the IRX10L2 gene. In the genome of two homologous chromosomes, the nucleotide sequence of the IRX10L3 gene in sequence 8 of the sequence listing has the following changes: In the genome of Nipponbare rice, a nucleotide T is inserted between nucleotides 460 and 461 in sequence 8 of the sequence listing, which corresponds to the insertion of a single nucleotide T between nucleotides 111-112 in the coding sequence of the IRX10L3 gene shown in sequence 7 of the sequence listing. This results in a frameshift mutation. The translation of the IRX10L3 protein terminates prematurely after 38 amino acids, thus knocking out the IRX10L3 gene.

[0151] IRX10 polygenic homozygous mutants Tm1, Tm2 and Tm3 were transplanted into a greenhouse for cultivation, and individual plants were harvested to obtain T1 generation seeds.

[0152] Example 2: Application of IRX10 protein in regulating crop growth

[0153] 2.1 Phenotypic observation of IRX10 gene-related mutants

[0154] T1 generation seeds of the IRX10 multi-gene homozygous mutants Tm1, Tm2, ​​and Tm3 obtained in Example 1, as well as wild-type Nipponbare, were sown in the field, with 24 plants per treatment. Phenotypic observation was conducted after three months of growth. The results are as follows: Figure 2As shown in Figure A, compared to the wild type, the plant height of the IRX10 mutant materials was significantly shorter: the plant height of the IRX10 polygenic homozygous mutant Tm1 was 69.3±1.0 cm, the plant height of the IRX10 polygenic homozygous mutant Tm2 was 38.2±0.6 cm, the plant height of the IRX10 polygenic homozygous mutant Tm3 was 30.6±1.0 cm, and the plant height of the wild-type Nipponbare (WT) was 105.8±0.9 cm. This indicates that the IRX10, IRX10L, IRX10L2, IRX10L3, IRX10L4, and IRX10L5 proteins and their encoding genes can regulate rice plant height. Knocking out the IRX10, IRX10L, IRX10L2, IRX10L3, IRX10L4, and IRX10L5 genes in rice resulted in shorter rice plant height.

[0155] 2.2 Detection of xylose content in IRX10 gene-related mutants and wild-type plants

[0156] IRX10 gene-related mutant and wild-type plants (japonica rice variety "Nipponbare") grown for 3 months were collected, with at least 10 main tillers from each material, and the xylose content in the cell walls was measured. The specific procedures are as follows:

[0157] 2.2.1 Preparation of ethanol-insoluble substances

[0158] Select the second internode below the panicle of rice plants that have grown for 3 months, dry them until their weight no longer changes, grind them into powder using a tissue homogenizer, and pass them through a 200-mesh sieve to obtain a uniform powder. Weigh 100 mg of the powder into a 10 mL centrifuge tube, and obtain the ethanol-insoluble material (AIR) of plant cell walls through the following steps: wash with 70% ethanol solution until the supernatant is colorless, wash with chloroform:methanol (1:1, v / v) solution until the supernatant is colorless, add 1 mL of acetone, and dry in an oven. Add 0.1 M sodium acetate solution (pH 5.0) and amylase (purchased from Megazyme) and amylopectin (purchased from Megazyme) to the AIR powder, and react in a shaker at 37°C for more than 24 hours to remove starch. Wash twice with distilled water, resuspend in 1 mL of acetone, and dry in an oven. Obtain the starch-free AIR sample.

[0159] 2.2.2 Monosaccharide content detection

[0160] Approximately 2 mg of starch-removed AIR powder was weighed, with five replicates for each sample. 40 μg of inositol was added to each replicate as an internal standard. The mixture was acid-hydrolyzed with 2 M trifluoroacetic acid and heated at 121 °C for 90 minutes to extract monosaccharide components. The extract was then reduced with sodium borohydride solution, neutralized with glacial acetic acid, washed with acetic acid / methanol (1:9, v / v) and methanol solutions, and dried. Acetic anhydride and pyridine were added, and the mixture was heated at 121 °C for 20 minutes to initiate a derivatization reaction. After drying, the mixture was extracted with ethyl acetate and distilled water. The upper ethyl acetate phase was collected, and the cell wall monosaccharides were separated and their content determined using gas chromatography-mass spectrometry.

[0161] See results Figure 2 In the data, B and WT represent wild-type plants, containing 238.0 ± 6.5 μg xylose per mg of ethanol-insoluble cell wall; in mutant bc18, the content is 187.2 ± 2.0 μg xylose per mg of ethanol-insoluble cell wall; in mutant Tm1, it is 117.1 ± 4.5 μg xylose per mg of ethanol-insoluble cell wall; in mutant Tm2, ​​it is 151.5 ± 6.3 μg xylose per mg of ethanol-insoluble cell wall; and in mutant Tm3, it is 64.9 ± 3.0 μg xylose per mg of ethanol-insoluble cell wall. These results demonstrate that, compared to wild-type plants, the xylose content in the cell walls of IRX10 gene-related mutant materials is significantly reduced.

[0162] Example 3: Application of IRX10 protein in regulating plant vascular development

[0163] 3.1 Catheter observation of IRX10 gene-related mutants

[0164] 3.1.1 Observation of duct shape

[0165] The second internode below the spike of IRX10 gene-related mutant and wild-type Nipponbare plants were taken after 3 months of growth. The stems were transversely cut with a Gillette blade, and the xylem vessels were observed under a microscope.

[0166] The results are as follows Figure 3 As shown in Figures A, B, and C, the vascular bundle area of ​​the wild-type Nipponbare plant is 1211.8 ± 27.4 μm. 2 The duct area of ​​mutant bc18 is 1058.5 ± 15.8 μm. 2 The duct area of ​​mutant Tm1 is 716.6 ± 14.3 μm. 2 The duct area of ​​mutant Tm2 is 668.2 ± 15.5 μm. 2 The duct area of ​​mutant Tm3 is 486.5 ± 11.9 μm. 2 The results showed that the duct area in both single and multiple mutants of IRX10 was significantly smaller, and its shape changed from approximately circular to irregular shapes such as ellipse or polygon.

[0167] 3.1.2 Observation of duct cell wall thickness

[0168] Further, transmission electron microscopy was used to observe the cross-section of the stems of the above materials. The stems were longitudinally cut into 2 mm sections and fixed with 4% paraformaldehyde. The sections were then embedded in aqueous resin and prepared into 90 nm sections using an ultramicrotome. After staining with 2% uranium acetate, the thickness of the vascular bundle cell walls was observed using transmission electron microscopy.

[0169] The results are as follows Figure 3 As shown in Figures D and E, the duct cell wall thickness of wild-type Nipponbare was 0.57±0.13 μm, the duct cell wall thickness of mutant bc18 was 0.27±0.06 μm, the duct cell wall thickness of mutant Tm1 was 0.16±0.06 μm, the duct cell wall thickness of mutant Tm2 was 0.25±0.08 μm, and the duct cell wall thickness of mutant Tm3 was 0.16±0.05 μm. The results indicate that the duct cell wall thickness of both single-mutant and multi-mutant mutants of IRX10 was significantly reduced.

[0170] 3.2 Observation of duct pits in IRX10 gene-related mutants

[0171] 3.2.1 Observation of vessel pit morphology

[0172] To further analyze the differences in the fine structure of the vessels in the mutants, a 1 cm length of the above-mentioned stem material was taken, immersed in a 10% peracetic acid solution, and reacted at 80°C for more than 12 hours. The vessel cells were then separated by stereomicroscopy, stained with DirectRed 23 dye, and observed by fluorescence microscopy.

[0173] The results are as follows Figure 4 As shown, the vessel pits of the wild-type Nipponbare plant are uniform in size, with a pit area of ​​0.84 ± 0.02 μm. 2 The pit area of ​​the vessels in mutant bc18 is 2.46 ± 0.04 μm. 2 The pit area of ​​the vessels in mutant Tm1 is 3.65 ± 0.06 μm. 2 The vessel pit area of ​​mutant Tm2 is 3.28 ± 0.06 μm. 2 The vessel pit area of ​​mutant Tm3 is 4.47 ± 0.13 μm. 2 The results showed that the duct pit area of ​​the single and multiple mutants of IRX10 showed an increasing trend to varying degrees, and two pits fused together appeared.

[0174] 3.2.2 Distribution of xylan in the vessels

[0175] The ductal cells were isolated according to the method in 3.2.1, and immunolabeled with xylan-specific antibody LM10 diluted at a ratio of 1:50 for 2 hours. Then, they were incubated with horseradish enzyme-labeled goat anti-rat for 2 hours. Finally, they were observed using a laser scanning confocal microscope.

[0176] The results are as follows Figure 5 As shown, in wild-type Nipponbare, xylan signals are mainly deposited at both ends of vessel pits, while in the bc18 mutant, xylan signals are significantly reduced; in the Tm1 mutant, xylan signals are almost absent. These results indicate that the decreased xylan content in both single and multiple mutants of IRX10 leads to the loss of specificity in the xylan nanolattice structure at both ends of vessel pits.

[0177] 3.3 Determination of ductal function in IRX10 gene-related mutants

[0178] To clarify the impact of differences in duct pit shape and structure, we further analyzed the duct function of IRX10 gene-related mutants.

[0179] 3.3.1 Rhodamine Transport Experiment

[0180] Wild-type Nipponbare, bc18, and Tm1 mutants were cultured in medium for two weeks until the four-leaf stage. They were then transferred to a solution containing 0.5 mM Rhodamine B and incubated for 3 hours. The base 2 cm of the second leaf was powdered and extracted with ethanol. The extract was analyzed using a microplate reader.

[0181] The results are shown in Figure 6. The vascular transport capacity of wild-type Nipponbare was 94.55 ± 0.47 nmol.g -1 The ductal transport capacity of the bc18 mutant is 55.82 ± 9.64 nmol.g -1 The ductal transport capacity of the Tm1 mutant was 5.14 ± 0.27 nmol.g -1 The results showed that the absence of xylan nanolattice structure around vessel pits in IRX10 gene-related mutants affects the water transport capacity of vessels.

[0182] 3.3.2 Determination of transpiration potential

[0183] Wild-type Nipponbare and IRX10-related mutants were cultured in the field for 3 months. Transpiration potential of leaves was measured every two hours from 9:30 AM to 5:30 PM using a photosynthesis meter. The results are shown in Figure 7. The transpiration potential of leaves fluctuated with time, reaching its highest at 11:30 AM. During this time period, the transpiration potential of wild-type Nipponbare leaves was 9.65 ± 0.87 mmol·m⁻¹. -2 ·s -1The leaf transpiration potential of the bc18 mutant was 7.10 ± 0.40 mmol·m⁻². -2 ·s -1 The leaf transpiration potential of the Tm1 mutant was 5.16 ± 0.70 mmol·m⁻¹. -2 ·s -1 The leaf transpiration potential of the Tm2 mutant was 4.75 ± 0.85 mmol·m⁻². -2 ·s -1 The leaf transpiration potential of the Tm3 mutant was 3.34 ± 0.59 mmol·m⁻¹. -2 ·s -1 The results showed that the absence of xylan nanolattice structure around the pits of the vessels in the IRX10 gene-related mutant affected the transpiration capacity of the plant leaves, indicating that the water transport capacity of the vessels had changed.

[0184] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. sequence list <110> Institute of Genetics and Developmental Biology, Chinese Academy of Sciences <120> Application of IRX10 protein in regulating plant vascular development and crop growth <160> 12 <170> SIPOSequenceListing 1.0 <210> 1 <211> 1254 <212> DNA <213> Rice (Oryza sativa L.) <400> 1 atgaggaggt gggtcttggc cattgccatt cttgctgctg ctgtatgctt cttccttgga 60 gctcaggccc aggaggtgcg gcagggccac cagacagaga ggatctcagg aagtgctggt 120 gatgtgttgg aagatgaccc tgttgggagg cttaaggtct atgtctatga tctcccaagc 180 aagtacaaca agaagctgct gaagaaggat cctaggtgcc tgaaccacat gtttgccgct 240 gagattttca tgcatcggtt cctgttgtca agcgctgtcc gaacttttaa tcccgaggaa 300 gctgattggt tctacacacc ggtgtacact acatgcgacc tgactccctc cggtcttccc 360 ttgcccttca aatccccaag aatgatgcgc agcgcaattg agctgattgc aacaaattg 420 ccttactgga atagatcaga gggggctgat catttctttg ttacaccaca tgactttggc 480 gcttgcttcc actatcagga agaaaaggca attggacgtg gaatcctccc attgcttcag 540 cgtgccaccc tggttcagac ctttggacaa aagaaccatg tctgcttgaa ggacggctcg 600 atcaccattc cgccatatgc acccccacag aaaatgcagg ctcatcttat tcccccagac 660 acccctcggt ctatctttgt atatttccgt ggtctgttct acgataccag caatgatcct 720 gagggtggat actatgcaag aggtgcccgc gcgtcggtttt gggagaattt caagaacaac 780 ccgctgtttg acatctcaac cgatcaccca cccacgtact acgaagatat gcagagatct 840 gtgttctgct tgtgcccatt gggctgggct ccatggagcc ccagactggt ggaagctgtg 900 gttttcggtt gtattccggt gatcattgca gatgacattg tcctcccctt tgctgatgct 960 atcccctggg aggagattgg cgtgtttgtc gccgaggagg atgttccgaa gctggacagt 1020 atcctgacat ccataccaac agatgttatc ctgaggaagc agaggcttct cgcgaacccg 1080 tcgatgaagc aggccatgct gttcccccag cctgctcagg caggagatgc attccatcag 1140 atactgaatg gtctcgctcg caagcttcca catggcgaaa acgtcttctt gaagcccggg 1200 gagagggccc tgaactggac tgctggaccg gtgggcgacc tgaagccttg gtag 1254 <210> 2 <211> 3023 <212> DNA <213> Rice (Oryza sativa L.) <400> 2 cttgaagcct gcttatatgt ggagttggag gaagaaggaa ggaagcctag gaaagcctcc 60 cctcaccttc tcaactcacc aaacgcttct tggtaggaga attgccagaa aagtttcccc 120 cttctccttg gatcctttaa ttacctcctg gcaattgcct gctagctcat aaggccggac 180 agggagaggg ttggagagga gaggagacac aggtgctggg aagaggcctg tggggatcaa 240 gaagatgagg aggtgggtct tggccattgc cattcttgct gctgctgtat gcttcttcct 300 tggagctcag gcccaggagg tgcggcaggg ccaccagaca gagaggatct caggtggggg 360 gccttgttt ttcttgtgtt attctcttct tggttttgat cttgatggat acattctttgt 420 cagttagtag aactcaaccc aagctgctta tttgtgcatc taagaaactg ttaaagcaaa 480 gcatttgaa atcatttagc ctgtcaattt gatttgtttg tggtcagacc tctccaccct 540 aaatatatag gatctatttc agatcagacg caaatggagg ttgtttatgt tggtgtatta 600 ttctaaatga tttggttgag ttatggttca atgcttggat tcagtttaaa ccttgtttta 660 ttgaacatct actggtattg atacacctgt tgatggtagg cattctttct tattggattt 720 tgtggtcctc gagacatctg ctgccaaaaa acttgaatgc cattctgtcc tctgaacctt 780 caaatatttt tcctcctctt catcttcttt atgatatgca gtatcatgtt ctaggcacc 840 aatatgataa gaacatagcc attagcctaa ttatgtttca tttaatctag cataggaaca 900 cacttacttt atccctgtag gttgcttagg tatcatacct tgatcccaag ctatgcttct 960 aaatttaga atattcttt ccctaattct gtgcaaggtc tatagatact ccatagcttt 1020 tgcttaatat tgtggcgaaa caaatgcagg aagtgctggt gatgtgttgg aagatgaccc 1080 tgttgggagg cttaaggtct atgtctatga tctcccaagc aagtacaaca agaagctgct 1140 gaagaaggat cctaggtgcc tgaaccacat gtttgccgct gagattttca tgcatcggtt 1200 cctgttgtca agcgctgtcc gaacttttaa tcccgaggaa gctgattggt tctacacacc 1260 ggtgtacact acatgcgacc tgactccctc cggtcttccc ttgcccttca aatccccaag 1320 aatgatgcgc agcgcaattg agctgattgc aacaaattgg ccttactgga atagatcaga 1380 gggggctgat catttctttg ttacaccaca tgactttggc gcttgcttcc actatcaggt 1440 acatgcacaa gtacaaaacc ttgctgttgg acacatttat tcctccttta gtgagcaaat 1500 tctggacttt gagcattagt ttagtactga acacttaagt ttgcaacagc tgggacattt 1560 ctaattgcag cagagaacat ggggagcaat ggggggcatc aagatgctag attcactgta 1620 ttaatattat tagttggcaa ctctgattta attggaacac ttcagtttgc acatgttctt 1680 tgtccctttg tagaatataa atatttttca gaaaaaaata ttttctctca acctatttaa 1740 atgtcacact aaggtcagat tcatattcat gattaatcat ctaccctgat tggcaccttc 1800 tatcatactt tgtgcccaca ttctctttgg ttatttcagc gtattgatct tttgcttctg 1860 ttattgtttc aggaagaaaa ggcaattgga cgtggaatcc tcccattgct tcagcgtgcc 1920 accctggttc agacctttgg acaaaagaac catgtctgct tgaaggacgg ctcgatcacc 1980 attccgccat atgcaccccc acagaaaatg caggctcatc ttattccccc agacacccct 2040 cggtctatct ttgtatattt ccgtggtctg ttctacgata ccagcaatga tcctgagggt 2100 ggatactatg caaggtaaat tttccgctta tctccaatac atcctatcat ctgctcattg 2160 gctcgtagca tgtggagtct gatgtttctg tatgtcctat cagaggtgcc cgcgcgtcgg 2220 tttgggagaa tttcaagaac aacccgctgt ttgacatctc aaccgatcac ccacccacgt 2280 actacgaaga tatgcagaga tctgtgttct gcttgtgccc attgggctgg gctccatgga 2340 gccccagact ggtggaagct gtggttttcg gttgtattcc ggtgatcatt gcagatgaca 2400 ttgtcctccc ctttgctgat gctatcccct gggaggagat tggcgtgttt gtcgccgagg 2460 aggatgttcc gaagctggac agtatcctga catccatacc aacagatgtt atcctgagga 2520 agcagaggct tctcgcgaac ccgtcgatga agcaggccat gctgttcccc cagcctgctc 2580 aggcaggaga tgcattccat cagatactga atggtctcgc tcgcaagctt ccacatggcg 2640 aaaacgtctt cttgaagccc ggggagaggg ccctgaactg gactgctgga ccggtggggcg 2700 acctgaagcc ttggtagcat gcatctcact gcctatagaa atgtaaaaac ttttttgaaa 2760 aagatatatt tacagacatg atatatactt gcgccttgag atcaccatgg ttgcatagtg 2820 ctgaccataa atcaacaaat gtacattttt ttgtatcttt ttccctgata gctatatacc 2880 atatatcttc attccttttc ttcccattag cggatgaaat gccggcattt gtccgctttg 2940 tccatatatg tggataactg gatatcataa tgaaattatg agatttaatt catgctgaat 3000 gcgtgacata atttcaaatg tgg 3023 <210> 3 <211> 1248 <212> DNA <213> Rice (Oryza sativa L.) <400> 3 atggcgatgc ggctgtcttc ggcggccgtg gcgctcgcgc tgcttctggc cgccaccgcc 60 ttggaggacg ttgccagggg ccaggacacc gagaggatcg aagggagtgc gggtgatgtg 120 ctggaagatg accctgtcgg gaggcttaag gtgtatgtct acgagttgcc cactaaatac 180 aacaagaaga tggtggcaaa agattccaga tgcctcagcc acatgtttgc tgcagagatt 240 ttcatgcatc gattcctctt gtcaagtgcg atccggactt taaatccgga ggaagctgat 300 tggttctata ctccagtata cacgacgtgc gatcttactc catggggtca tcccttgccc 360 ttcaagtctc caaggattat gagaagtgca attcagttca tttcctcgca ttggccctat 420 tggaacagga cagatggggc agaccatttc tttgttgtgc cacatgattt cggagcatgc 480 ttccattatc aggaagagaa ggccattgag cggggaatcc tgccattgct tcgccgtgcg 540 acgctggtcc agacttttgg gcagaaggat catgtctgcc tcaaggaagg gtccatcaca 600 atcccaccat atgctcctcc tcagaagatg aaaactcacc ttgttccccc agagacacct 660 cgatcaatct ttgtctattt ccgtggttta ttctatgata ccgcaaatga tcctgagggt 720 ggttactatg caagaggtgc ccgtgcatcg gtgtgggaga acttcaagaa caatcctctt 780 tttgacatct ccacagacca cccaccaacc tactacgagg acatgcagcg ttccatcttc 840 tgcttgtgcc ctctaggctg ggctccgtgg agcccccggt tggttgaagc cgttgtgttc 900 ggctgcattc cagtgatcat tgcagacgac attgtcctac cttttgcgga cgcgatcccg 960 tgggatgaga tcggtgtatt tgtggcggag gacgacgtcc caaagctgga caccatcctg 1020 acatcgatac caatggatgt gatcctgcgg aagcagaggc tgctagcgaa cccgtcgatg 1080 aagcaggcca tgctgttccc gcagcccgcc cagccaggtg acgccttcca ccagatactc 1140 aacgggctgg gacggaagct tccacacccg aagagcgtgt acctggaccc ggggcagaag 1200 gtgctgaact ggacccaggg cccggtgggt gacctgaagc catggtag 1248 <210> 4 <211> 3496 <212> DNA <213> Rice (Oryza sativa L.) <400> 4 agcttccagc caccggccac cgccatcatc atcctcgcga gctcgcctcc ctcctctccc 60 ctccccgtgt tcctctcctt cctcctcctc ctcctcgatt ggccagccgc atccagcatc 120 cttcttgggg gtttcgtcga ggtcggtggt tggaagggag gagaggtggg gggagccatg 180 gcgatgcggc tgtcttcggc ggccgtggcg ctcgcgctgc ttctggccgc caccgccttg 240 gaggacgttg ccaggggcca ggacaccgag aggatcgaag gtacgcaccg cacgcgctcg 300 cgagcggttc aagtagcgat ttctttttct ttttcttctt ttttttggcg atggttgatt 360 gcttgctccg ttgctcgatg cgtgcttttc gcaatgtttg tgtcggtgca tttgatccgt 420 ggttgctgct gcattctgat ggctggaatc tgaccaatct gttgtgggga ttgagtgaag 480 cgagcagttt aatttagggc gtatcttgca gataatcact tgcctcccgc gaccttgggg 540 acaaggggat ctgctgttgc ttggatatca actgttaatc gtagcactat ttgttctagg 600 tccaatttcc agttaatata aacttccccg gggaccggga gagtctgcta ctagctactt 660 acttcatgcc caacacgcag ccacctctgc tagtagtaga ttcatgtgtt atccaacatc 720 tttggtttag cctctacgcg atcgttccgc gttccatgtt ccgcggcctc gattcagttg 780 ccatggaata attatgctc atgtgtgagt gtgatgtgtg gttgagtgta gctcaagctc 840 gattctttct caattactcc ggtcaaaact cgatgcataa tgaggtgaac agtaagcatg 900 tgaatcaatg tagtattagt tgatatataa gcaagtggct gaaattactg aactataccg 960 acgactatca tactcggtca atctactgga tctgatgagt ttctcaaata gaattgacat 1020 gtgctttcgc attcttgcat tcctttttta tgtcattttt ttaacgattt tttgagtcat 1080 tttatgcaa tataacata ctagtagtac taaaaaaact ctggttggca agctactcct 1140 aagtcttatc cactccataa ctaattgccg cagtttgact tatgatttgc ttaccaaaaa 1200 gagaaactta tgggaagctg ttaggcctgt caaattatgg tggctgctag gacactacat 1260 aattctcact ttttcattgt acatatgacc atatcttagg gagtgcgggt gatgtgctgg 1320 aagatgaccc tgtcgggagg cttaaggtgt atgtctacga gttgcccact aaatacaaca 1380 agaagatggt ggcaaaagat tccagatgcc tcagccacat gtttgctgca gagattttca 1440 tgcatcgatt cctcttgtca agtgcgatcc ggactttaaa tccggagga gctgattggt tctatactcc agtatacg acgtgcgatc ttactccatg gggtcatccc ttgcccttca 1560. agtctccaag gattatgaga agtgcaattc agttcatttc ctcgcattgg ccctattgga 1620. acaggacaga tggggcagac catttctttg ttgtgccaca tgattcgga gcatgcttcc attatcaggt aaatcaagga atcattaacc ttgaactatt gtatatcaaa tcatggaac attack cctcatctac attack caacgaatc attcagctcc aatgtacca ttgttaccct tttacatgtt caaccatggg atcttattg catttggttt ttccatgatt caggaagaga aggccattga gcggggatc ctgccattgc ttcgccgtgc gacgctggtc cagacttttg ggcagaagga tcatgtctgc ctcaaggaag ggtccatcac aatcccacca 2040. 2040. 2040. 2040. 2040. 2040. 2040. 2040. 2040. 2040. 2040 tttgtctatt tccgtggttt attctatgat accgcaaatg atcctgaggg tggttactat gcaaggtgag caccaagaac atgcaatcac tcgatgcttc ctcataagca ttgttgatga attattgtaa cccaaaagtc ttgatgaacc tgctttatgc aatgcagagg tgcccgtgca 2220 tcggtgtggg agaacttcaa gaacaatcct cttttgaca tctccacaga ccacccacca 2280 acctactacg aggacatgca gcgttccatc ttctgcttgt gccctctagg ctgggctccg 2340 tggagccccc ggttggttga agccgttgtg ttcggctgca ttccagtgat cattgcagac 2400 gacattgtcc taccttttgc ggacgcgatc ccgtgggatg agatcggtgt atttgtggcg 2460 gaggacgacg tcccaaagct ggacaccatc ctgacatcga taccaatgga tgtgatcctg 2520 cggaagcaga ggctgctagc gaacccgtcg atgaagcagg ccatgctgtt cccgcagccc 2580 gcccagccag gtgacgcctt ccaccagata ctcaacgggc tgggacggaa gcttccacac 2640 ccgaagagcg tgtacctgga cccggggcag aaggtgctga actggaccca gggcccggtg 2700 ggtgacctga agccatggta ggtcagatgc acccacccac catacatagt tgctgctgtc 2760 ctcgcttgtt tgtgcgcaca gcacattctt cagaagatca gatgagagac tgatgatgat 2820 cagtaaaaga aaaccaagcc aatccgttaa atgtaaagcc tcctttcttc cttcctccta 2880 ctacctttgc ttctttttga gacgatgtgc cctgcctgcc ctgcgccctt gccagtgaca 2940 ttcccttggt ttttatgttg ttgttgctgt agtagtatta gacaactggt acctataggc 3000 gatgaaggtg ttgtatcata tacttaagtg ctacctccgt tttttaatag atgacgccgt 3060 tgactttttc tcacatgttt gacccttcgt cttattcaaa aaatttacgt aattataatt 3120 tattttgtta tgagttgttt tatcattcat agtactttaa gtgtgattta tatcttatac 3180 atttgcataa aatttttgaa taagacgaat ggtcaaacat gtaagaaaaa gtcaacggcg 3240 tcatctaata aaaaacggag agagtagtac tagtaagttg atgatttatg acgggcaaaa 3300 aatatgtaag gttatatcac aacattcaga gccccaatcc agatggatgg atggattccc 3360 ctgcctgtgt gctgcactgt ttgcaaggct gcagaagatt tggtgcttgt gttgttcctg 3420 ggcttgcaca gatgtttcat gtatccaaag tcctgggtga agtgaaatgg atgagataag 3480 gaaaaaaaga agtcca 3496 <210> 5 <211> 1254 <212> DNA <213> Rice (Oryza sativa L.) <400> 5 atggggacga ggaggaggag tgcgcgagcg cgagcgcggc cgccgctggc catgccgctc 60 gccgtgcttc ttctcttcgc ctgttcttcc ggcgtcgcag ccgcggccgc tcagggcatc 120 gagcggatca aagacgatcc tgttggaaag ctgaaggtgt atgtctacga gctgcccccc 180 aagtacaaca agaacattgt ggcgaaagat tccagatgcc tcagccacat gtttgccaca 240 gagatcttca tgcatcgctt cctgctgtcg agtgcgatcc ggacttcgaa tccggacgaa 300 gccgattggt tctacactcc agtgtacacc acatgcgacc tcacgccatg gggccatccc 360 ctgaccacca agtctccacg catgatgaga agtgcgatca agttcatctc caagtactgg 420 ccctactgga acagaacgga gggtgcggat catttcttcg ttgtgccaca tgactttgca 480 gcgtgcttct acttccagga ggcgaaggct attgagcgag gcatccttcc ggtgctgcgc 540 cgtgctacgc tggtgcagac gtttgggcag aagaaccatg cgtgtctcaa ggatggctcc 600 atcacagtcc cgccgtatac tcctgctcac aagatcagag ctcatcttgt tccaccagag 660 actcctcggt caatctttgt ctacttccgt ggtttgttct atgatacttc gaatgatcct 720 gagggtggtt actatgcaag gggcgcccgt gcatcggtgt gggagaactt caagaacaac 780 ccgatgttcg acatctctac ggaccacccc caaacctact acgaggacat gcagcgtgca 840 gtcttctgcc tgtgcccgct gggctgggcg ccatggagcc ctcgtctggt ggaggccgtg 900 gtgttcggct gcatcccggt gatcatcgca gacgacatcg tcctcccctt ctccgacgcg 960 atcccgtggg aggagatcgc cgtgttcgtg gccgaggacg atgtcccgca gctggacacc 1020 atcctgacct ccataccaac ggaggtgatc ctccggaagc aggcgatgct cgcggagccg 1080 tcgatgaagc agaccatgct gttcccgcag cctgcggagc ccggggatgg cttccaccag 1140 gtgatgaacg cgctggcgag gaagctgccc catggcaggg atgtgttcct caagcccggg 1200 cagaaggtgc tcaactggac cgaggggacc cgggaagacc tgaagccgtg gtag 1254 <210> 6 <211> 3449 <212> DNA <213> Rice (Oryza sativa L.) <400> 6 caactattct attctgaatt ctcccatgcc ttctcctggt aaagaataat cccaccacct 60 cccctcta ctacctctcc ctcccccaa actcgagcct ccatctccct cccgatcca 120 gacaccgatc gaggcgttga cgacgacgtc tagctcatgg ggacgaggag gaggagtgcg 180 cgagcgcgag cgcggccgcc gctggccatg ccgctcgccg tgctttctct cttcgcctgt 240 tcttccggcg tcgcagccgc ggccgctcag ggcatcgagc ggatcaaagg tatgtgcgtc 300 tgttgctccg gtaatccaat taattacgcg ccgtcggttg ttgcatagat ctgaggaatc 360 acacgcgtgc tatgcccctc gtcgcgttt tggtggccgg aatttgattc ggtcgagtgc 420 tctctgctat gctatcgcac tgaacagagt gctccaaaat cttgagatcg aatctattta 480 tttcatctat gtgatgcgtc gtcaatcgat tatattatat atataatcct gacttgaata 540 tgctgagcag ctgaccataa atgaagtttg attaattcttt ttccatgtag caagacttct 600 atgctatcgt agctgttcca ttactttttc tttgcctgct ttctcgctac tccctcggcc 660 acaaaaatta gataatcttt gcttacttat tgatccaaaa acaagttcat cattaattat 720 tggtccaaaa acaagttcat cattaagtaa tcattttatt agaatttgtg aaagtagtga 780 acagatgtat tcgaagtaga taaaatagca tacaacttta ttaggatttg ataaaatgga 840 gggtattcta gtcttttctt ttatatctat tggtatgtgt gggatggccg gattgtttaa 900 aaatggagtt attttgggat ggatgaagtg ctagtatagc ttcaataatt atgctttgac 960 tgaagaatta aaaagaacac tttcagcaat gttttaactc catttatatt aactttcctt 1020 gctcgcggat atgattaaaa ttcacgaagt gctgctggtg atgtgctaga gaagacgatc 1080 ctgttggaaa gctgaaggtg tatgtctacg agctgccccc caagtacaac aagaacattg 1140 tggcgaaaga ttccagatgc ctcagccaca tgtttgccac agagatcttc atgcatcgct 1200 tcctgctgtc gagtgcgatc cggacttcga atccggacga agccgattgg ttctacactc 1260 cagtgtacac cacatgcgac ctcacgccat ggggccatcc cctgaccacc aagtctccac 1320 gcatgatgag aagtgcgatc aagttcatct ccaagtactg gccctactgg aacagaacgg 1380 agggtgcgga tcatttcttc gttgtgccac atgactttgc agcgtgcttc tacttccagg 1440 tgagtcatgt acaaggttcg gattcggaaa attttcagta attttgaact ccattgagaa 1500 gaaattattt agactgcaaa ctttagtttt ttgtggatta gcaaaatttg tgttaactcg 1560 gtcaaaattt gttcaatctt caaattgtat gggccttggt catacatagt gccataaacc 1620 atgaatcatc ctctcctata cactccctcc atttgaaaaa tataagtatt tttaattcaa 1680 aatgcgtagt tctaaggcga taatttcaat gtgcgggaat ccatgtattt tcagccaatc 1740 agatgcttgg gataggaatt taagcaattc aaaatttgaa atttgactaa taaaatgact 1800 aaaaacagaa gttttccgtt aaccttgatc tttgctagac aatcagcaaa tgctcatatt 1860 ttataacgga gagagagtgt attattaccc atgtaacgtg gaatatgcaa actcatagaa 1920 tggaaattca tcctagtaac gaagccacac acaacacctg ttatttatcc cttactcatg 1980 tttattcgct tctagcatca tgtacatcca acgtgcattc agatcaacat gttcctactt 2040 ttcaggaggc gaaggctatt gagcgaggca tccttccggt gctgcgccgt gctacgctgg 2100 tgcagacgtt tgggcagaag aaccatgcgt gtctcaagga tggctccatc acagtcccgc 2160 cgtatactcc tgctcacaag atcagagctc atcttgttcc accagagact cctcggtcaa 2220 tctttgtcta cttccgtggt ttgttctatg atacttcgaa tgatcctgag ggtggttact 2280 atgcaaggta tgcacctaat taaattatgc agctgaaaag ctgtaggtct taatcaatca 2340 ttgattagaa agtcttgata cacctaccat taggttctat attgtgtaaa gcctaaggga 2400 attgatggat ctgaagtagc agaaattcag gaatattttt gccctccaaa ttttgaaatt 2460 acatgataac aaaatttgaa taaattttgc aaaattaaaa aaaaaaagtt gctgggtgag 2520 gggaagggtg gcaaggtggt tgaaatttca taccaaaatg ttgaatcctg cctaacatat 2580 gcaatgcagg ggcgcccgtg catcggtgtg ggagaacttc aagaacaacc cgatgttcga 2640 catctctacg gaccaccccc aaacctacta cgaggacatg cagcgtgcag tcttctgcct 2700 gtgcccgctg ggctgggcgc catggagccc tcgtctggtg gaggccgtgg tgttcggctg 2760 catcccggtg atcatcgcag acgacatcgt cctccccttc tccgacgcga tcccgtggga 2820 ggagatcgcc gtgttcgtgg ccgaggacga tgtcccgcag ctggacacca tcctgacctc 2880 cataccaacg gaggtgatcc tccggaagca ggcgatgctc gcggagccgt cgatgaagca 2940 gaccatgctg ttcccgcagc ctgcggagcc cggggatggc ttccaccagg tgatgaacgc 3000 gctggcgagg aagctgcccc atggcaggga tgtgttcctc aagcccgggc agaaggtgct 3060 caactggacc gaggggaccc gggaagacct gaagccgtgg tagatagata tatacagtat 3120 atgatagtgt tatggcacca ttgccattca ccatggctgg tgttcttgtg taaatgacac 3180 caccggatga gaagctgatc aatgaggata tattgatgat gatgagcgaa ccacaccatt 3240 gaatgtaaag ctccctttcc ttgcttcttt ttaaggccat gtgcctgtat ggcaatgtct 3300 gttttgacca ttgttgtagt gttagcgtat tacaggtcat ggattatgta atactagtca 3360 ataacccata cgttacaccg tgcaacttgc acaggctatt tgttcatata tatataaaac 3420 ataaatagta aaattttgtc ataggcaga 3449 <210> 7 <211> 1269 <212> DNA <213> Oryza sativa L. <400> 7 atggggacga ggccgtgcgc cggcgtggcg tcggcggtgg cggcggcggt ggccgttctg 60 ctgctcgccg tgtcttgctt cgccgcggcg gcgacgacga cgcagaagca tggcaggatg 120 tcagggaagg gcggcgacgt gctggaggac gacccgacgg ggagctgaa ggtgttcgtg 240. 240. 240. 240. 240. 240. 240. 240. 240. 240. 240 cacatgttcg cggcggagat cttcatgcac cagttcctcc tctccagccc cgtgcgcacc 300 ctcgacccgg aggaggccga ctggttctac acacccgcct acaccacctg cgacctcacc 360 ccgcagggct tcccgctccc cttccgcgcc ccgcgcatca tgcgcagcgc cgtccgctac 420 gtcgccgcca cgtggcccta ctggaaccgc accgacggcg ccgaccactt cttcctcgcg 480 ccgcacgact tcggcgcctg cttccactac caggaggagc gcgccatcga gcgcggcatc 540 ctcccggtgc tccggcgcgc cacgctggtg cagacgttcg gccagcgcca ccacccctgc 600 ctccagccgg ggtcgatcac cgtgccgccg tacgccgatc cccgcaagat ggaggcgcac 660 cgcatcagcc ccgcgacgcc gcgctcgatc ttcgtgtatt ttcgggggct tttctacgac 720 atggggaatg atcccgaggg cgggtactac gcccgcggcg cgaggcgtc ggtgtgggag 780 aacttcaagg acaacccgct gttcgacatc tcgacggagc acccggcgac gtactacgag 840 gacatgcagc gggccatctt ctgcctgtgc ccgctggggt gggcgccgtg gagcccccgg 900 ctggtggagg cggtggtgtt cgggtgcatc ccggtgatca tcgccgacga catcgtgctc 960 cccttcgcgg acgccatccc gtggggggag atcagcgtgt tcgtggcgga ggaggacgtg 1020 ccgaggctgg acaccatcct ggcgtccgtc ccgctcgacg aggtcatccg gaagcagcgg 1080 ctgctggcgt ccccggcgat gaagcaggcc gtgctgttcc accagccggc gaggcccggc 1140 gacgcgttcc accagatact caacgggctc gcgcgcaagc tgccgcaccc caagggcgtc 1200 ttcctcgagc ccggcgagaa gggcatcgac tgggaccagg gcctcgagaa cgacctcaag 1260 ccatggtag 1269 <210> 8 <211> 3938 <212> DNA <213> Rice (Oryza sativa L.) <400> 8 tttttctact cttccccttg cttcgcttcg ctcatcagct tgaccgcctc acctcaccac 60 cccaagctag acgacctcgt cttcctcctc cacctcctcg cgacctcgca ttgttagatc 120 agcctcgatt catctccaat ctttcggcgc cgctcccgga gagagggaca gcggcgcagg 180 cggtgagcac gaggaggagc agagctaatc aaaagccctc tcgccgctcg agctctcgtg 240 caccgtgccg ccgccgccga cgccgacgct gacgccgacg gcttaattca gtcttgtcgg 300 ctacatctct tggcgtgctg aaattggtgg gagcacgaga gcggcgccaa tggggacgag 360 gccgtgcgcc ggcgtggcgt cggcggtggc ggcggcggtg gccgttctgc tgctcgccgt 420 gtcttgcttc gccgcggcgg cgacgacgac gcagaagcat ggcaggatgt caggtacgct 480 ccgcctcacc agcaacctac ctaccttagc tagataaata gatagataag caactactga 540 cctacctgct gccaattcct cgtctcctcc gcttggattc ttgattccat gggatgggtc 600 accgtcgatg gcgtctcaaa ttgacacccc tcgagattca gatttaccgg cttgcgtcac 660 cgccgatgaa ttcgcaaaat gtcactctct attgctcatc ctgcattcgc gaagcatgct 720 tgcaattcat ccaatccatt catacgctct cataaaattc tcctctgctt cacaattcac 780 acatccgatt ctttcggttc cattccacca ctacaaaatc agcttcagtt cttgctttct 840 tggatcaata tgcagttata caaattatcc tgcaataaca atccagtagc tagctagcta 900 gcttccctgt gaccaacca tcaaaccagg ctgtgatgag tgatccaacc atgtggccag cacaggccac caacttttat caagaacctg tactacatgc tgcagcagca gtaactgtaa ctgtaatac agcaatgcca cccagtaacc accaacctgc ctgcctgcct gcctgtaact gcttgcagct gcagtgagct caccagaatt ggttggtagg tgaagccttt aggacagctt cttggtgaca tgatcacact actacattgc atcgcatttg catcaatggc tgcttctagt 1260. taggtcacca ccaccatgct tgatggctca gatctctcct tcttcttcgt ccccaaagtt ttcccttttc tgtaccatct cctttgtgat cacgcatcca tagtgcggct gcctcagga 1320 tcgaggcaaa ggctgccatt attgccgtct cttgtgagca gcgcctgtgc ctaacttgca caacttgttc ttctttaatc tgcaaattag agcctgcgag atctggaaat ttgttagtat aaaaagtgaa aaaagtgtgc tcgattgagc agtggtcgcc cacttactgg cctgtgaatg attctgtttc ttcttcttcg tcctttttttt ttcttgaact ggacatgcta acaaccaacc 1560. tggttggtga ttaacaatag ataaacataa aactccatga aagtagacag gagttgatag ggattgattt ttagtagtat agcacaagat cctatggttg gtgaactact gactgcaact 1680 aattcggtca cttccaccaa ctaatatgct cactaatact agtgattagt tatattgtac 1740 tgttatgaac tgaagttggt gcagacgtgc agttgttctg aaactgataa tctatacccc 1800 aacctgaaca tctggcagca cattttgctg ttgtgaactc aaaaggcagt tctgaaactg 1860 aacatgtata atccacaaca attaatttgg gctcatttta ctgttcttta ctgaaactaa 1920 tcatgtaaac caccatcctg aacatattgc acattcctgc tgctttgaac tgaaaagatg 1980 gtgctctgat ctaagaaggc attgctgaaa ctgaccctgc aaaacacaca aacatttaat 2040 tttactctct ttctttttct ggaactcttc aggtttgctg tttatgaact gaaaaactga 2100 cgctgaaact gatcatgtgg atctctagct acaatgtagt tgtagggtaa ttatattaca 2160 attatactat ggttatactg taaggttttt tcaccttaaa aaacttgcaa cagtttttta 2220 gcaattccca aacccccaac ctgagcatct ggcacatttt tatgctctga tcccaaaagg 2280 cagcctgaaa ctgatactgc aaaaacgaca tgttactact cgtttgctga acattgtccg 2340 catttcggca tgtacttcct ccctccgtcc caaagtataa gaattagaga ggcctattca 2400 gattcgtagc actaggaaat gtccgatccg atccgtttca aggtttacca tggcaaatca 2460 ataccacgaa caactaacaa gcagtgaaaa ttaactgatg gatgtggatg tggattgtgc 2520 agggaagggc ggcgacgtgc tggaggacga cccgacgggg aagctgaagg tgttcgtgta 2580 cgagatgcca cgcaagtaca acctgaacct gctggccaag gacagccggt gcctgcagca 2640 catgttcgcg gcggagatct tcatgcacca gttcctcctc tccagccccg tgcgcaccct 2700 cgacccggag gaggccgact ggttctacac acccgcctac accacctgcg acctcacccc 2760 gcagggcttc ccgctcccct tccgcgcccc gcgcatcatg cgcagcgccg tccgctacgt 2820 cgccgccacg tggccctact ggaaccgcac cgacggcgcc gaccacttct tcctcgcgcc 2880 gcacgacttc ggcgcctgct tccactacca ggaggagcgc gccatcgagc gcggcatcct 2940 cccggtgctc cggcgcgcca cgctggtgca gacgttcggc cagcgccacc acccctgcct 3000 ccagccgggg tcgatcaccg tgccgccgta cgccgatccc cgcaagatgg aggcgcaccg 3060 catcagcccc gcgaccgccg gctcgatctt cgtgttttt cggggcttt tctacgacat 3120 ggggaatgat cccgagggcg ggtactacgc ccgcggcgcg agggcgtcgg tgtgggagaa 3180 cttcaaggac aacccgctgt tcgacatctc gacggagcac ccggcgacgt actacgagga 3240 catgcagcgg gccatcttct gcctgtgccc gctggggtgg gcgccgtgga gccccccggct 3300 ggtggaggcg gtggtgttcg ggtgcatccc ggtgatcatc gccgacgaca tcgtgctccc 3360 cttcgcggac gccatcccgt ggggggagat cagcgtgttc gtggcggagg aggacgtgcc 3420 gaggctggac accatcctgg cgtccgtccc gctcgacgag gtcatccgga agcagcggct 3480 gctggcgtcc ccggcgatga agcaggccgt gctgttccac cagccggcga ggcccggcga 3540 cgcgttccac cagatactca acgggctcgc gcgcaagctg ccgcacccca agggcgtctt 3600 cctcgagccc ggcgagaagg gcatcgactg ggaccagggc ctcgagaacg acctcaagcc 3660 atggtagccg ccatggatga tgagctatat acacacagat tatagagacg agacatggat 3720 tggatcgcga gagatgtaca ttgattggtt cgtttcttct gattaagccg ctgttaattt 3780 cttcttcttt ctttcttttt tctagttttt tttccttgtt gttttttctt tcaaggaggc 3840 gatccgtgtt catgtattgt gcaattgtgc gtgcttaatt aatcgttgca atgtgcacga 3900 tttcttctat agtgaaatta gccttttccc tttttgtc 3938 <210> 9 <211> 1305 <212> DNA <213> Rice (Oryza sativa L.) <400> 9 atggtgggcg caagggctgg gcgggtccca gcggcggcgg cggcggcggc tgcggtgctc 60 atcgtcgctg cgtgcgtctt ctcgtcgctg gcgggggccg ccgccgccgc cgaggtggtc 120 gggggagcgg ctcaggggaa caccgagcgc atctcaggaa gtgctggtga tgtgctagaa 180 gacaatccag ttgggcggtt gaaggttttt gtttatgact tgccaagcaa gtataacaag 240 aggatagtcg ccaaggatcc ccgatgcctc aaccacatgt ttgctgctga gatatttatg 300 catcgtttct tgctctctag tgctgttcga acgctcaacc ctgaacaggc tgattggttc 360 tatgcccctg tttacactac ttgtgatttg actcatgctg gacttccatt gccattcaag 420 tctcctagga tgatgcggag cgcaatccag tttctttcaa gaaaatggcc attctggaat 480 agaactgatg gagcagatca ttctttgtt gttccacatg attttggggc atgctttcac 540 tatcaggaag agaaagctat tgagcgtggg attacktccat tgctccgccg tgccacattg 600 gttcaaactt ttggacagaa gaaccatgtg tgcttgaagg agggttctat cactatacca 660 ccatatgcgc ctcctcagaa aatgcaagct cactgattc ctcctgatac tccccgttca 720 atctttgttt acttcagagg cctgttctat gataggga atgaccctga gggtggggtat 780 tatgcaagag gtgcccgagc ttcactgtgg gagaatttca agaacaatcc cctcttcgac 840 atttctacgg aacaccagc tacttactt gaagatatgc agcgctcagt cttctgcttg 900 tgccccttgg gatgggcacc atggagccct agatggtgg aggctgtt ttcggttgc 960 attccagtca tcatagctga tgatattgtg cttccgttg ctgacgcaat cccatgggac 1020 gaaatcggtg ttttgttga cgaggaggat gttccaaggc tagactcaat cctcacatcc 1080 atcccaattg atgatatcct aaggaagcaa agattactcg ccaatccatc atgaacag 1140 gccatgttgt ttccgcagcc agcacaacct agagatgcat tccatcagat cctcaacggc 1200 cttgctcgga agctcccaca cccagacagt gtatacttaa agccaggcga gaagcatctc 1260 aactggactg cggggcctgt agcagatctg aaaccttgga aatag 1305 <210> 10 <211> 3555 <212> DNA <213> Oryza sativa L. <400> 10 ccccccacca ctttgccatt ttcccttctc tcttccccct ttttttgctg cctccctttg 60 cttcctcctc cagcgatcaa gctctcgcga cttctagaag cttccacggc gtctcccccc 120 cgaggcattt ctggtgggtg gagcgggggg gcggcgctca tggtgggcgc aagggctggg 180 cgggtcccag cggcggcggc ggcggcggct gcggtgctca tcgtcgctgc gtgcgtcttc 240 tcgtcgctgg cgggggccgc cgccgccgcc gaggtggtcg ggggagcggc tcaggggaac 300 accgagcgca tctcaggtga gcgagctgta cttctgcacc cgattcgttc gacttgctgg 360 gttgttagaa tgttagcttg tggattgtga atttgtgatg cgagtgattc agttgtagcc 420 tcttgtgttg gatttgttca cgaacttggc taggggaatc gctcaactgg ctcagcttgt 480 ttgtaaatat ttttcccttt acctgattca aagttgggat gctgttgcta ggcgaggacg 540 tgtcagccta acaccactca gttcttaggc cgaaattctc ctctaatcta gtagtagtac 600 ttcaaccatg gatattcata tgcaattgca agttcgacct gatatgtgc attgtacaac 660 aacacccgtg ctcttagaat cgcacattcg cactgatatg cttttcccc cctataaaaa 720 aggaaatccg gatgtccaat actagtttga tgtaaattca gtactgactg caatggatcg 780 ctgattctcg tgcttatcga aggaatgtaa tctctccgcg ccagcaagg gttgtgtga 840 catatagaaa tgttgaatta ccattagca cctatttctc ctcctctcc gtttcttta 900 cttgttacat agttacatt tgatagtttt aagacaatca tatatgcttg tgctacctgc 960 agacctacac ttgttatgtc attgtctcat tgacaagtc ctatccaact aacgttagtt 1020 attgtcaaga tactgtgatc tgcttggagc caatccaatt gtgcatataa cttttccgat 1080 aatggaattg gatccaatta ttgttctat ttaatacaat attccagga taaacagtta 1140 ccccataaat gcaattgtgc caaaaag agaggaat ctacctgagt ttttaggag 1200 agtggtgcat tctgagttta tgttcagag aaacagaa tttcgttca attattcatg 1260 atttattgaa aatgaatttcc cctaagcttg tattgagttt tacattcttt cttcttgtta 1320 acatttattt ccaccttaaa tattacttat taattatatg gggaatatcc aatgaaaacc 1380 atcaatag cggaaacccg tgaaaaccat acctaaaact ttctaaatt catgaaaaaa 1440 attackagata tagctataga catgaaatac attcatacca attctcaagt ccaacttag 1500 cttcattga gagaacaa aaagagaat tctaggtgaa tagtgtcatg ttaccattca 1560 cccaaaatgt cttttctgtt actcctaaat aagttgagt ttgaacttga tattttggtgg 1620 gatgtattt catgcctaca tccatctca gtattttt tcatgaattt atgaaacttt 1680 taggttaggt ttcatgggtt tcacttatt tgatggttt cgctggatat gtccccttat 1740 tatataa cttcatgact gttttcttg gatctgctt atcagtttta tgagatagt 1800 actatgtgct tctactcaac cavatcatt gacctgcagg aagtgctggt gagtgctag 1860 aagacaatcc agttgggcgg tgaagtttt tgtttatga cttgccaagc aagtataaca 1920 agggatagt cgccaaggat ccccgatgcc tcaccacat gttgctgct gagatattta 1980 tgcatcgttt cttgctctct agtgctgttc gaacgctcaa ccctgaacag gctgattggt tctatgcccc tgtttacact acttgtgatt tgactcatgc tggacttcca ttgccattca agtctcctag gatgatgcgg agcgcaatcc agtttctttc aagaaaatgg ccattctgga atagactga tggagcagat catttctttg ttgttccaca tgattttggg gcatgctttc actatcaggt aatgatatat tggtttattt gtcttcttac ctgccaagtc cttgtcactg ttgtagctag tcatgatttt agttccacaa tagttttaag aaataggtag ctcaggatag atcctacctt tagtccctat ctattctgaa actgcatatg caatggttat atgtctttac aggaaga agctattgag cgtgggatac ttccattgct ccgccgtgcc acattggttc aaacttttgg acagaagac catgtgtgct tgaaggaggg ttctatcact attackcaccat atgcgcctcc tcagaaaatg caagctcact tgattcctcc tgatactccc cgttcaatct ttgtttactt cagaggcctg ttctatgaca atgggaatga ccctgagggt gggtattatg 2700. 2700. 2700. 2700. 2700. 2700. 2700. 2700. 2700. 2700 ttattgactc tgttgaactt gttatacaga ggtgcccgag cttcactgtg ggagaatttc 2760 aagaacaatc ccccttcga catttacg gaacacccag ctacttacta tgaagatatg 2820 cagcgctcag tctctgctt gtgccccttg ggatgggcac catggagccc tagattggtg 2880 gaggctgtag tttcggttg cattccagtc atcatagctg atgatattgt gcttccgttt 2940 gctgacgcaa tcccatggga cgaatcggt gttttgttg acgaggagga tgttccaagg 3000 ctagactcaa tcctcacatc catcccatt gatgatatcc taaggaagca agattactc 3060 gccaatccat caatgaaca ggccatgttg ttccgcagc cagcacaacc tagagatgca 3120 ttccatcaga tcctcaacgg ccttgctcgg aagctcccac acccacag tgtatactta 3180 aagccaggcg agaagcatct caacggact gcggggcctg tagcagatct gaaccttgg 3240 aaatagaaaa gctcacatga aaaactcggc ttgatgtta tgtgaagtt ttaagttatg 3300 tccttttcca ctgaggagtt tctagtttca catagctaga tgatatttat caagttgta 3360 tatctccaat gtcgctgagc taatgccta ctctgtggaa gtgagtggaa cagtttgtag 3420 ctttgtaatt gtataaaaaa aattaaagca atgaatgtca gaatttgttc ttgtgttcac 3480 taaactagct gggtgacccg cgcaattgcg cggctagcac ccatagaaag ttatatatat 3540 tttaatatga tttta 3555 <210> 11 <211> 1263 <212> DNA <213> Rice (Oryza sativa L.) <400> 11 atgggatcaa gaacggtggg gtggtggcta ctggcggcgg ccgtcgtgct cgccgcggcg 60 gcggcggatt ccggggaggc ggagcgcgcg gcagagcagc acagcgagcg catctcaggg 120 agtgccggcg atgtgctcga agacaatcct gtggggaggt tgaaggtctt catctatgac 180 ctcccaagaa agtacaacaa gaagatggtc aacaaggatc cccggtgcct caatcacatg 240 tttgctgcag aaatattcat gcatcgcttc ttgctctcga gtgctgttcg gacactcaac 300 cccaaggagg ctgattggtt ttacacgcca gtttatacta cttgtgacct aactccggct 360 ggactgccct tgccatttaa gtcgccgagg gtgatgagga gtgcgatcca gtacatttcg 420 cacaagtggc ccttctggaa tagaacagat ggagcagacc acttctttgt tgtcccgcat 480 gattttggtg catgttttca ctatcagga gaaaaagcta ttgagcgtgg aattctaccg 600. ttgctgcaac gtgctacgtt ggtccaaaca tttggacagg agaccatgt ttgcttaaaa gagggttcta tcactatacc accctacgct cctccacaga aaatgcaggc tcacttgatc 660 ccccctgaca ctccacgttc aatcttcgtc tacttccggg gactattcta tgacactgga 720 aatgaccccg agggtggtta ctatgcaaga ggggcgcgag cttccctatg ggaaaacttc 780 aagaacaatc cattattcga catttccacg gatcaccctc ccacctacta tgaagacatg cagcgtgctg tcttctgtct gtgcccattg ggctgggcac catggagccc taggttggtt900 gaggctgtgg tctttggctg cattccagtc atcattgctg acgacattgt gctaccattt 960 gccgatgcaa tcccttggga ggaattggc gtgtttgttg aggagaga tgttccaaag ttagacacca tcctcacatc gatgccaatc gacgatattt tagagca aagattgctt gcaaatccat caatgaagca ggccatgttg tttccacagc cagcccaacc aagagatgcg ttccaccaga tcttgaatgg ccttgctcgc aagcttccac acccagaggg agtatacttg caacccagtg acaagcgcct caactggact gctggacctg ttggagatct gaaagcttgg 1260 tag 1263 <210> 12 <211> 4127 <212> DNA <213> Rice (Oryza sativa L.) <400> 12 cccctctttc tctctcttcc ctcgatttgt cccaaccaaa tcttcttggc ttcatccatc 60 cttgctccac gtcccaatct cctcctcctc ctcctcccca cttcttcttc tttgtttttt 120 ctcttttttt ttcccgaatt cccagaccaa atctcctcgc tttttgcttc gttctccgct 180 ccggtgaccc ggaggaggcg gaggttggag aaagtttgcg cctttgcggt gagtggtggt 240 gtcggtgatg ggatcaagaa cggtggggtg gtggctactg gcggcggccg tcgtgctcgc 300 cgcggcggcg gcggattccg gggaggcgga gcgcgcggca gagcagcaca gcgagcgcat 360 ctcaggttat ctctctcaag accgattttt tctttctttt tttttcacat agaaaagagg 420 ggtgcttttt tagatgctgg gaaccttgat gtgtgctgat gctattaacc aagatgtgtg 480 gtggtatgat ttgcttaatc tagtgtgccg atttgggagg agggtatgtt agttcttggc 540 tgttgcttgg agttgagagg acttaggtca atcgtttgca ggctatctga gattcagttg 600 gagatctctt cttgtgccat ccattgtatt ccttttttag tttacagtcc aagtatgtta 660 ctgtcaaagt tgtggcctat gtgcatcatc atcatcatcc ccatccactg caaagttcca 720 agctgaggtt gtaggttcta ttggtggttg ttccccgttt ccttgttaat tttcttgatt 780 ttgttacctc cctgccgcta agattgctta tgcggtaaca tgcaattttt gatggtgata 840 tttgttcctt tgattggatt tcatggatcc aacaagagga acataactca atcacaaatt 900 tgatcaaatc caagtagcac cgtgccgttc catggttcaa ttcatcatgt tcaagtgata 960 agtacttcac agctctcaag aaagtgataa aacacttaac ttggtattac tgctggtata 1020 gtttcacaaa tatagttaag agtcttagag atgggttcat gcttcgttgc tgtgaaactt 1080 aattgtttca tatttagcga ttgtgttatt tctaaaccat caattgttaa attgtaggga 1140 gtgccggcga tgtgctcgaa gacaatcctg tggggaggtt gaaggtcttc atctatgacc 1200 tcccaagaaa gtacaacaag aagatggtca acaaggatcc ccggtgcctc aatcacatgt 1260 ttgctgcaga aatattcatg catcgcttct tgctctcgag tgctgttcgg acactcaacc 1320 ccaaggaggc tgattggttt tacacgccag tttatactac ttgtgaccta actccggctg 1380 gactgccctt gccatttaag tcgccgaggg tgatgaggag tgcgatccag tacatttcgc 1440 acaagtggcc cttctggaat agaacagatg gagcagacca cttctttgtt gtcccgcatg 1500 attttggtgc atgttttcac tatcaggtgt gtgctaattc ccttggcatc tcacatttgg 1560 atttgtctgt gtgtaattct ctttctgatt tcattgttta ctttggagtt tggacatacc 1620 cttttagatg tcaacgctaa ggataaaaca ttattttgac cttttctact cacaagttct 1680 gaagtagtgt tattaactgt tgtatactat ttgtaggaag aaaaagctat tgagcgtgga 1740 attctaccgt tgctgcaacg tgctacgttg gtccaaacat ttggacagga gaaccatgtt 1800 tgcttaaaag agggttctat cactatacca ccctacgctc ctccacagaa aatgcaggct 1860 cacttgatcc cccctgacac tccacgttca atcttcgtct acttccgggg actattctat 1920 gacactggaa atgaccccga gggtggttac tatgcaaggt tagcaatgtt tttaaagtaa 1980 cagatctgaa tttttatcct tctggttttg tcccatgtac tcacttcagt gatgttatac 2040 agaggggcgc gagcttccct atgggaaaac ttcaagaaca atccattatt cgacatttcc 2100 acggatcacc ctcccaccta ctatgaagac atgcagcgtg ctgtcttctg tctgtgccca 2160 ttgggctggg caccatggag ccctaggttg gttgaggctg tggtctttgg ctgcattcca 2220 gtcatcattg ctgacgacat tgtgctacca tttgccgatg caatcccttg ggaggaaatt 2280 ggcgtgtttg ttgaggagaa ggatgttcca aagttagaca ccatcctcac atcgatgcca 2340 atcgacgata ttttaagaaa gcaaagattg cttgcaaatc catcaatgaa gcaggccatg 2400 ttgtttccac agccagccca accaagagat gcgttccacc agatcttgaa tggccttgct 2460 cgcaagcttc cacacccaga gggagtatac ttgcaaccca gtgacaagcg cctcaactgg 2520 actgctggac ctgttggaga tctgaaagct tggtagtgag cagcttccct ctccttggac 2580 gatccttgta tgaaaacctc gtagggtcag tcatcagttc atacttcctg aaatgtgatg 2640 taatcaacgt gcagtgccgg gtgtaatgta caccttggcc ttatctaact taatgaatct 2700 ttgtaagtta gtaaattttg aaagaaatga aaattgtatc gttggaatca gaatggcaag 2760 ttagatcagt ggttattttg gttgccaagt ttttgcagca taccctcata catatattgg 2820 cgaagaactg gtggcaagaa tgctgaactg cacatcaggt aagtcacaag gttgtaagta 2880 acattagatg aatgaagatg tacatttttc atgtttttgg acagatatgc ttagattctg 2940 ccttttacgg atacatccat gctattttgc aagccatgta actgatttat aattaatgca 3000 gcctctccgg tggttccatg acgatcatcg cattactcgt tatcatcatc agccaggtcg 3060 aactgcttga gcagcgcgcc gcagatgggc agtgggatgt cttgaggta gccgagtggg 3120 ttggctaagc gcaggttgtc acagatgccg acattcctgg tgagcaaaac acgggcacga 3180 tccctttcgg cctcgatttc agagcagttt gcaagggggc tgtggacgag gacaacctcg 3240 cagatgtcct cctcatggct gtccttgagc tcaatctttgt atgtcccggt ccatcagtc 3300 acaccgtcaa tcgcacgctc gaccttgtcg gtgccaaagt gcttgcactc cagcctgacc 3360 ttggcacctg caatcaagag atcaggtgag ccaatgctgg acttctaacc aggttagcgt 3420 ttcatgttta caatttgatg cacaaactct gcagtttctt tggtcttcct aagttctgct 3480 gctcatatat tcggagatta ttcatttcag catttgcatt gcagactcct gacccaaaaa 3540 agaatttcct tacccttgat atactcggtg acattggtct cgaacccggc gcggcacgtg 3600 tcacagtaga cacggccttg gacgacgtag tccggtgcgt cggtggcagt ggcagtgctg 3660 gcaaggacat agaagaggat gccgaagatc accggaatgg tgcggagaga ggccatggat 3720 gatgtggctg caaatgtgtt tttccccttc ctctattatt atttggggaa aatgtgagag 3780 gagggtactg aaatggcctg gtttggaacg gattcgagct gtggtataag taacctgggg 3840 aacctggagg atttccatgg ctgtccttgc aagtaggaat cgttggagat gccgttggga 3900 tcgagtggct gaagagaagg cagattgttg atttgatcct acttttgatg atgaccaagc 3960 taaaaatggg tgaatgtttt gacgagcgag gctattatgc cgttgtgggt gttttagcca 4020 gccagaaatc tgtcgaattt tgacgttacg gaagcagcag acaacttggg aacctgaata 4080 gatttcatgt tgggcgcctt gaatgcagta acttcatagg atgttca 4127

Claims

1. Application of substances that regulate the expression of protein-coding genes in reducing plant height and / or inhibiting plant vascular development; The protein is IRX10-IRX10L-IRX10L3, and IRX10-IRX10L-IRX10L3 is composed of IRX10, IRX10L, and IRX10L3. The IRX10 is a protein encoded by a DNA molecule whose coding sequence is shown in SEQ ID No. 1; The IRX10L is a protein encoded by a DNA molecule whose coding sequence is shown in SEQ ID No. 3; The IRX10L3 is a protein encoded by a DNA molecule whose coding sequence is shown in SEQ ID No. 7; The substance that regulates the expression of protein-coding genes is any one of the following: B1) A nucleic acid molecule that inhibits or reduces the expression of the protein-coding gene; the nucleic acid molecule is a DNA molecule that expresses an sgRNA targeting the protein-coding gene or is an sgRNA targeting the protein-coding gene, wherein the target sequence of the sgRNA is nucleotides 1918-1937 of sequence 2 in the sequence listing, nucleotides 1325-1344 of sequence 4 in the sequence listing, and nucleotides 444-463 of sequence 8 in the sequence listing; B2) Expression cassettes, recombinant vectors, or recombinant microorganisms containing the nucleic acid molecules described in B1); The plant in question is rice.

2. A method for reducing rice plant height, comprising knocking out the gene encoding the protein of claim 1 in the rice genome to obtain rice with reduced plant height.

3. The method according to claim 2, characterized in that: The gene encoding the protein described in claim 1 in the rice genome that is knocked out is formed by inserting nucleotide A between nucleotides 1934 and 1935 in sequence 2 of the rice genome sequence listing, inserting nucleotide C between nucleotides 1340 and 1341 in sequence 4, and inserting nucleotide T between nucleotides 459 and 460 in sequence 8.

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