CN108864266B

CN108864266B - Protein SSH1 related to rice graininess and grain type as well as encoding gene and application thereof

Info

Publication number: CN108864266B
Application number: CN201810757562.9A
Authority: CN
Inventors: 谭禄宾; 蒋丽芸; 刘凤霞; 付永彩; 顾凭; 朱作峰; 孙传清
Original assignee: China Agricultural University
Current assignee: China Agricultural University
Priority date: 2018-07-11
Filing date: 2018-07-11
Publication date: 2020-08-18
Anticipated expiration: 2038-07-11
Also published as: CN108864266A

Abstract

The invention discloses a protein SSH1 related to rice graininess and grain type, and a coding gene and application thereof. The plant phenotype related protein SSH1 and the coding gene thereof provided by the invention can regulate and control the plant phenotype: the silent strains (RNAi-SSH1-1 and RNAi-SSH1-2) are basically consistent with the phenotype of the mutant SSH1, and show that the dropping property is reduced and the grains are lengthened; t is₁The generation SSH1 gene complementation strains (GC-SSH1-1 and GC-SSH1-2) and the over-expression strains (OE-SSH1-1 and OE-SSH1-2) show increased grain fall and shortened grains. The result shows that the protein SSH1 related to the plant falling-grain property and/or the grain type and the coding gene thereof can regulate the plant falling-grain property and/or the grain type. The invention has important significance for rice breeding.

Description

Protein SSH1 related to rice graininess and grain type as well as encoding gene and application thereof

Technical Field

The invention relates to the field of plant genetic engineering, in particular to a protein SSH1 related to rice grain falling property and grain type, and a coding gene and application thereof.

Background

Rice (Oryza sativa L.) is one of the most important and the earliest domesticated food crops, and rice is eaten by about more than half of the global population. With the annual increase of the world population and the annual decrease of the available cultivated land area, the contradiction of unbalanced food and food supply and demand relations is increasingly prominent. Therefore, it is imperative to breed more rice varieties with high, stable and high quality.

The shattering property is one of important agronomic traits influencing the yield establishment of crops, and is always the key point and the focus of crop genetic research, and the reduction or the loss of the shattering property is one of key events in the crop domestication process. The seeds fall naturally after maturity, which has important ecological significance for the offspring propagation, however, in the process of crop domestication, people tend to select varieties with reduced falling property for further cultivation so as to avoid yield loss and improve crop harvesting efficiency. The results of genetic studies show that rice grain shattering is a typical complex quantitative trait controlled by multiple genes.

The grain type is also an important factor affecting rice yield and is composed of grain length, grain width and grain thickness. In domestication breeding, the grain type is also one of the characters selected manually, and provides an excellent model for domestication research.

Both rice grain shattering and grain type are important agronomic traits affecting yield. Therefore, the gene for further separating and controlling the rice grain dropping property and the grain type not only provides reference for clarifying the molecular regulation mechanism of the grain dropping property and the grain type, but also is beneficial to cultivating a new rice new variety with high yield and moderate grain dropping property by a molecular breeding method.

Disclosure of Invention

The invention aims to provide a protein SSH1 related to rice graininess and grain type, and a coding gene and application thereof.

The protein provided by the invention is obtained from rice and named SSH1 protein, and is (a1) or (a 2):

(a1) a protein consisting of an amino acid sequence shown in a sequence 1 in a sequence table;

(a2) and (b) a protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in the sequence 1, is related to the plant seed shattering property and/or the seed type and is derived from the sequence 1.

In order to facilitate purification and detection of the SSH1 protein of (a1), a tag as shown in Table 1 may be attached to the amino terminus or the carboxy terminus of the protein consisting of the amino acid sequence shown in SEQ ID No. 1 of the sequence Listing.

TABLE 1 sequences of tags

The SSH1 protein of (a2) above may be synthesized artificially, or it may be obtained by synthesizing the coding gene and then expressing it biologically. The gene encoding the SSH1 protein of (a2) above can be obtained by deleting one or several codons of amino acid residues from the DNA sequence shown in sequence No. 4 of the sequence listing, and/or by carrying out missense mutation of one or several base pairs, and/or by attaching a coding sequence of the tag shown in Table 1 above to the 5 'end and/or 3' end thereof.

The gene encoding the SSH1 protein (SSH1 gene) also belongs to the protection scope of the invention.

The gene is a DNA molecule as described in any one of (b1) to (b5) below:

(b1) the coding region is a DNA molecule shown as a sequence 4 in the sequence table;

(b2) a DNA molecule shown as 2310-5997 th nucleotide of a sequence 2 in a sequence table;

(b3) a DNA molecule shown as 2310-5997 th nucleotide of a sequence 3 in a sequence table;

(b4) a DNA molecule which hybridizes with the DNA sequence defined in (b1) or (b2) or (b3) under stringent conditions and encodes a protein associated with the seed shattering and/or the seed type of a plant;

(b5) and (b) DNA molecules which have more than 90% of homology with the DNA sequences limited by (b1) or (b2) or (b3) or (b4) and encode proteins related to the seed shattering and/or grain type of plants.

The stringent conditions can be hybridization and washing with 0.1 XSSPE (or 0.1 XSSC), 0.1% SDS solution at 65 ℃ in DNA or RNA hybridization experiments.

The recombinant expression vector, the expression cassette, the transgenic cell line or the recombinant strain containing the SSH1 gene belong to the protection scope of the invention.

The recombinant expression vector containing the SSH1 gene can be constructed using existing plant expression vectors. The plant expression vector comprises a binary agrobacterium vector, a vector for plant microprojectile bombardment and the like. When SSH1 gene is used to construct recombinant expression vector, any one of enhanced, constitutive, tissue-specific or inducible promoter can be added before its transcription initiation nucleotide, and can be used alone or in combination with other plant promoters; in addition, when the SSH1 gene is used to construct a recombinant expression vector, enhancers, including translational or transcriptional enhancers, may be used, and these enhancer regions may be ATG initiation codon or initiation codon of adjacent regions, etc., but must be in the same reading frame as the coding sequence to ensure proper translation of the entire sequence. The translational control signals and initiation codons are widely derived, either naturally or synthetically. The translation initiation region may be derived from a transcription initiation region or a structural gene. In order to facilitate the identification and screening of transgenic plant cells or plants, plant expression vectors used may be processed, for example, by adding genes expressing color-changing enzymes or luminescent compounds in plants, antibiotic markers having resistance, or chemical reagent-resistant marker genes, etc.

The recombinant expression vector can be specifically a recombinant expression vector obtained by replacing a fragment between BmaHI and SpeI enzyme cutting sites of the plasmid pCAMBIA1301 with a DNA molecule shown in a sequence 4 of a sequence table.

The invention also protects the application of the gene of the SSH1 protein or the SSH1 gene in regulating and controlling the seed shattering property and/or the seed shape of the plant.

The invention also protects a method for cultivating transgenic plants, which is to inhibit the expression of SSH1 gene in target plants to obtain transgenic plants; the transgenic plant satisfies the phenotypes of (c1) and/or (c2) as follows:

(c1) the seed shattering is lower than that of the target plant;

(c2) the kernel length is larger than that of the target plant.

The inhibition of the expression of the SSH1 gene in the plant of interest can be achieved by introducing an interference vector into the plant of interest.

The interference vector may be specifically an interference vector obtained by replacing a fragment between the SpeI and SacI cleavage sites of the plasmid pTCK303/JL1460 with a DNA molecule represented by the 1098 th to 1420 th positions from the 5 'end of the sequence table, and replacing a fragment between the BamHI and KpnI cleavage sites of the plasmid pTCK303/JL1460 with a reverse complement of a DNA molecule represented by the 1098 th to 1420 th positions from the 5' end of the sequence table.

The invention also provides a method for reducing the plant seed shattering and/or increasing the seed length, which comprises the following steps: reducing the expression and/or activity of SSH1 protein in a target plant, reducing the plant falling property and/or increasing the kernel length.

The invention also protects a method for cultivating transgenic plants, which is to introduce the SSH1 gene into a target plant to obtain a transgenic plant; the transgenic plant satisfies the phenotypes of (d1) and/or (d 2):

(d1) the seed shattering property is higher than that of the target plant;

(d2) the kernel length is less than that of the target plant.

The SSH1 gene is introduced into a target plant by any one of the recombinant expression vectors described above. The recombinant expression vector can be transformed into plant cells or tissues by conventional biological methods such as Ti plasmid, Ri plasmid, plant virus vector, direct DNA transformation, microinjection, conductance, Agrobacterium mediation and the like.

The invention also provides a method for increasing the plant seed shattering and/or reducing the seed length, which comprises the following steps: increasing expression and/or activity of SSH1 protein in a target plant, increasing plant shattering and/or reducing kernel length.

The invention also protects the application of the SSH1 protein, or the SSH1 gene, or any one of the methods in plant breeding.

The breeding aims to breed plants with low plant shattering and/or large grain length.

The breeding aims to breed plants with high plant shattering and/or small seed length.

Any of the above plants is a dicotyledonous plant or a monocotyledonous plant. The monocot plant can be a plant of the order gramineae. The plant of the order gramineae may be a gramineae. The gramineous plant may be a plant of the genus oryza. The plant of the genus oryza may specifically be rice, for example indica line YIL 100.

The plant phenotype related protein SSH1 and the coding gene thereof provided by the invention can regulate and control the plant phenotype: the silent strains (RNAi-SSH1-1 and RNAi-SSH1-2) are basically consistent with the phenotype of the mutant SSH1, and show that the dropping property is reduced and the grains are lengthened; t is₁The generation SSH1 gene complementation strains (GC-SSH1-1 and GC-SSH1-2) and the over-expression strains (OE-SSH1-1 and OE-SSH1-2) show increased grain fall and shortened grains. The result shows that the protein SSH1 related to the plant falling-grain property and/or the grain type and the coding gene thereof can regulate the plant falling-grain property and/or the grain type. The invention has important significance for rice breeding.

Drawings

FIG. 1 is a schematic genotype of indica rice line YIL 100.

Fig. 2 is a comparison of the phenotypes of mutant ssh1 and indica line YIL 100.

FIG. 3 is the cloning of the SSH1 gene.

FIG. 4 is a comparison of the relative expression levels of mRNA of the SSH1 gene in different tissues of mutant SSH1 and indica rice line YIL 100.

FIG. 5 shows phenotypic identification of transgenic plants.

Detailed Description

The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. In the quantitative tests in the following examples, three replicates were set up and the results averaged.

Indica rice line YIL100 is derived from an introgression line constructed by taking Yuanjiang ordinary wild rice (O.rufipogon Griff.) as a donor parent and taking excellent indica rice variety Teqing (Oryza sativa ssp. indica) as a receptor parent, and the phenotype of the introgression line is natural shattering. Both the yunnan yuanjiang common wild rice (o. rufipogon Griff.) and the elite indica rice variety ultragreen (Oryza sativa ssp. indica) are described in the literature: tan, l., Li, x, Liu, f., Sun, x, Li, C, Zhu, z, Fu, y, Cai, h, Wang, x, Xie, d., and Sun, c. (2008). Control of key transfer from product to yield growth in edge registration.nat. genet.40: 1360-. The construction method of indica rice line YIL100 is described in the following documents: tan L., Liu F., Xue W., Wang G., Ye S., Zhu Z, Fu Y, Wang X, and Sun C. (2007). Development of Oryza rugosa and Oryza sativa interrogsorosis and assessment for derived-related quantitative traci.JIntegr plantatBio.49: 871 884; a schematic genotype of indica rice line YIL100 is shown in FIG. 1.

Plasmid pCAMBIA1300 is described in the following documents: tan, l., Li, x, Liu, f., Sun, x, Li, C, Zhu, z, Fu, y, Cai, h, Wang, x, Xie, d., and Sun, c. (2008). Control of key transfer from product to yield growth in edge registration.nat. genet.40: 1360-.

Plasmid pCAMBIA1301 is described in the following documents: yu B., Lin Z., Lin H., Li X., Li J., Wang Y., Zhang W., Zhu Z., Zhai W., Wang X., Xie D., Sun C. (2007) TAC1, a majorgquantative train controlling tiller angle in plant J, 52: 891-cake 898. the public is available from the university of agriculture in China to repeat the experiment.

Plasmid pTCK303/JL1460 is described in the following documents: wang Z., Chen C., Xu Y., Jiang R., HanY., Xu Z., and Chong K. (2004). A Practical Vector for Efficient knock down of Gene Expression in Rice (Oryza sativa L.). Plant mol.Bio.Rep.22: 409) 417. the public is available from university of agriculture in China for the repetition of this experiment.

Agrobacterium EHA105 is described in: WuY, Fuy, ZhaoS, GuP, Zhuz, SunC, andTan L. (2015). CLUSTERED PRIMARY BRANCH 1, a new alloy of DWARF11, control personal architecture and seed analysis in plant Bio.J.14: 377-386 the public is available from the university of agriculture in China to repeat the experiment.

Example 1 obtaining of Gene SSH1 controlling Rice Fall characteristic and grain type

Indica rice line YIL100 is treated by Ethyl Methane Sulfonate (EMS) and forms a genotype homozygous mutation system after multi-generation auto-crossing and character observation. A mutant with reduced noisiness was found by screening of the mutant system and was designated ssh 1.

Comparative experimental results for the granularities of ssh1 and YIL100 are shown in FIGS. 2 at A, B and C. A comparison of kernel morphology for ssh1 and YIL100 is shown in fig. 2 at D, E, F and G. Compared with YIL100, the ssh1 mutant has reduced granularities (YIL100 has a particle size ratio of 78.97 + -8.93%, and ssh1 has a particle size ratio of 10.99 + -3.61%). Comparative experimental results for the granularities of ssh1 and YIL100 are shown in FIGS. 2 at A, B and C. A comparison of kernel morphology for ssh1 and YIL100 is shown in fig. 2 at D, E, F and G. Compared to YIL100, the kernel of ssh1 was longer (D and E in fig. 2).

Backcrossing mutant ssh1 with indica rice line YIL100, F₁The generation appeared as a falling grain, and therefore it was assumed that the gene controlling the mutation trait was recessive. Hybridization F₁Selfing to produce F₂In the progeny, the ratio of plants showing shattering to non-shattering was close to 3: 1(205 plants, 72 plants, none, chi)²＝0.10<χ² _0.05,13.84). Therefore, the reduction of ssh1 granularities was controlled by a recessive single gene, and the mutant gene was named the suppression of fusion 1 (ssh 1).

From F ₂20 dominant falling single plants and 20 recessive non-falling single plants are selected to respectively extract DNA, the two groups of DNA are respectively mixed in equal quantity to construct a dominant pool and a recessive pool, and an Illumina HiSeq2500 sequencing platform (Beijing Helikang biotechnology limited,item number BFC2013036) was re-sequenced and a SNP cluster linked to the no-shattering phenotype was detected on chromosome 7 using the MutMap strategy (a in figure 3). Further using 72 recessive individuals to position the target gene SSH1 between the SNP markers SNV4 and SNV8, wherein the physical distance between the two is about 3104Kb (B in figure 3), and 3 SNPs are identified on the basis of combining whole genome re-sequencing and SNP verification. According to rice reference genome (http:// rice. plant biology. msu. edu) annotation and real-time fluorescent quantitative PCR, it was found that only SNP marker SNV6 is located within the gene (the 3' end of the ninth intron of LOC Os07g13170 has a single nucleotide variation from base C to A, specifically shown in sequence 2 and/or sequence 3 at position 5782 in the sequence list, C and D in FIG. 3) and is expressed at the abscission site controlling seed shattering, so LOC _ Os07g13170 was used as the candidate gene and the gene was named SSH 1.

Wherein, the real-time fluorescent quantitative PCR uses rice ubiquitin gene (LOC _ Os03g13170.1 as internal reference gene, and the primers used for identifying SSH1 gene expression are as follows, P1, P2 and P3:

p1_ F: 5'-TGACTATGAGGACGACTTGA-3' (735 th and 754 th sites of the sequence 4 in the sequence table)

P1_ R: 5'-AGATGTACTTCTTGCCGAGC-3' (reverse complement of position 894-913 of sequence 4 in the sequence Listing)

P2_ F: 5'-ATGTCCCAGCATCCACATTT-3' (1234 th-1253 th site of sequence 4 in the sequence table)

P2_ R: 5 '-GGAGTAATGGCAAAGGGGAG 3' (reverse complement of 1387 th 1368 th site of the sequence 4 in the sequence listing)

P3_ F: 5'-GCCACCACCAGTTCTACTTC-3' (position 1472-1491 of sequence 4 in the sequence Listing)

P3_ R: 5 '-CTCTCTGGACAAACGACGAA 3' (reverse complement sequence at position 1569-1588 of sequence 4 in the sequence Listing)

The primers for detecting the ubiquitin gene are as follows:

Ubi_F:5’-CTGTCAACTGCCGCAAGAAG-3’；

Ubi_R:5’-GGCGAGTGACGCTCTAGTTC-3’。

the expression results of the SSH1 gene in different tissue sites of indica rice line YIL100 and mutant SSH1 are shown in figure 4, and when the P1 primer is used for detection, the expression level of the SSH1 gene in the two materials is not obviously different. The detection results of the primers P2 and P3 show that the expression of the SSH1 gene in the mutant is down-regulated in different tissues compared with the expression of the gene YIL100 in all the tissues, so that the SNP SNV6 is supposed to cause the change of the mRNA expression of the SSH1 gene. Wherein DP indicates developing rice ear, AZ indicates abscission, F indicates rice floret, DBP indicates days before flowering, and DAP indicates days after flowering.

The MutMap strategy described above is referenced in the following documents: (1) abe, a., Kosugi, s., Yoshida, k., Natsume, s., Takagi, h., Kanzaki, h., Matsumura, h., Yoshida, k., Mitsuoka, c., Tamiru, m., Innan, h., Cano, l., Kamoun, s., and Terauchi, R. (2012), Genome sequencing real environmental analysis report input vector logic in using mutmap.nat.biotechnol.30: 174-; (2) lu, h, Lin, T, Klein, j, Wang, S, Qi, j, Zhou, q, Sun, j, Zhang, z, Weng, y, and S, (2014). QTL-seq identification an early floor QTL located near floors T in the room, the. app. gene 127: 1491-1499; (3) takagi, H.A., Abe, A.A., Yoshida, K.K., Kosugi, S.A., Natsume, S.A., Mitsuoka, C.A., Uemura, A.A., Utsushi, H.A., Tamiru, M.S., Takuno, S.A., Innan, H.A., Cano, L.M., Kamount, S.A., and Terauchi, R.2013. QTL-seq: Rapid mapping of qualitative position location in rice by book genetic equation of DNA from tissue distributed locations J.74: 174. 183. A.

Primers were designed based on the SSH1 gene of indica 93-11 reference genome sequence (http:// www.gramene.org /) and flanking sequences flanking both sides thereof, and the sequences of the primers were as follows:

f: 5'-GGCTCCAAAGTATAAAACAC-3' (1 st to 20 th positions of the sequence 2 and/or the sequence 3 in the sequence table);

r: 5'-TCAATAGCATATGTATGCAA-3' (reverse complement of position 7181-7206 of sequence 2 and/or sequence 3 in the sequence Listing).

The whole genome DNA of indica rice line YIL100 and mutant ssh1 was extracted by CTAB method. The two DNAs are respectively used as templates to amplify DNA fragments with the length of about 7.2kb, and the sequences of the DNA fragments are respectively a sequence 2 and a sequence 3 in a sequence table.

The 1 st-2309 th sites of the

sequences

2 and 3 in the sequence table are the 5' flanking sequences of the SSH1/SSH1 gene of indica rice line YIL100 and mutant SSH1 respectively, and the 2310 st-5997 th sites are the SSH1/SSH1 gene sequences containing introns (2729 st-2830 th site, 2857 st-2978 th site, 3010 st-3092 th site, 3181 st-3291 st site, 3438 st-4051 st site, 4097 st-4186 th site, 4291 st-4386 th site, 4524 st-4963 st site, 5066 st-5784 th site is the intron sequences). The corresponding SSH1 gene CDS sequence without intron is shown in sequence 4 in the sequence table.

According to the alignment analysis of the sequencing results of the SSH1/SSH1 genes of the indica rice line YIL100 and the mutant SSH1, a single nucleotide variation from C to A exists at the 3' end of the ninth intron of the SSH1 gene, which is specifically shown as the 5782 th position in the sequence 2 and the sequence 3 in the sequence table.

The 2310-5997 th sites of the sequence 2 and the sequence 3 in the sequence table are sequences of SSH1 genes in the genomes of indica rice strain YIL100 and mutant SSH1 respectively; the sequence 4 in the sequence table is the cDNA sequence (CDS) thereof. The sequence 2, the sequence 3 and the sequence 4 in the sequence table are all proteins (SSH1 proteins) shown as a coding sequence 1.

Example 2 application of SSH1 Gene in regulating plant Seleteness and seed type

Firstly, construction of recombinant plasmid GC-SSH1

The fragment between KpnI and XbaI cleavage sites of the plasmid pCAMBIA1300 was replaced with a DNA molecule represented by the sequence 2 in the sequence table to obtain a recombinant plasmid GC-SSH1 (which was verified by sequencing).

Secondly, construction of recombinant plasmid OE-SSH1

The fragment between BmaHI and SpeI enzyme cutting sites of the plasmid pCAMBIA1301 is replaced by DNA molecules shown in a sequence 4 of a sequence table to obtain a recombinant plasmid OE-SSH1 (the sequencing is verified).

Third, construction of recombinant plasmid RNAi-SSH1

The fragment between the SpeI and SacI cleavage sites of the plasmid pTCK303/JL1460 was replaced with the DNA molecule shown from the 1098 th to 1420 th positions of the 5 'end of the sequence listing, and the fragment between the BamHI and KpnI cleavage sites of the plasmid pTCK303/JL1460 was replaced with the reverse complement of the DNA molecule shown from the 1098 th to 1420 th positions of the 5' end of the sequence listing, to give the recombinant plasmid RNAi-SSH1 (which was confirmed by sequencing).

Fourth, obtaining transgenic complementary plant

1. And (2) transforming the recombinant plasmid GC-SSH1 obtained in the first step into agrobacterium EHA105 by using a freeze-thaw method, transforming a transformed agrobacterium into a mutant SSH1 by using a method of infecting rice calluses with agrobacterium, performing 3 rounds of screening by using an NB culture medium containing hygromycin (the concentrations of the hygromycin used in each round of screening are 15mg/L, 25mg/L and 35mg/L), and performing pre-differentiation, differentiation and rooting to obtain a transgenic plant.

2. After step 1 is completed, extracting total DNA of transgenic plants, and performing PCR identification by using hygromycin resistance genes carried by a pCAMBIA1300 vector (the size of a target band of a positive plant is 1,254bp), wherein the primers are as follows:

Hpt_F:5’-TGCATCATCGAAATTGCCGT-3’

Hpt_R:5’-AAACGACGGCCAGTGAATTC-3’

through identification, 11 positive T strains are obtained₀The generation SSH1 gene complementation strain. Will T₀The seed produced by selfing of the generation SSH1 gene complementation strain is named as T₁Generation complementary seeds of T₁The rice plant grown by generation complementary seeds is named as T₁The generation SSH1 gene complementation strain. Two complementation lines were randomly selected and named GC-SSH1-1, GC-SSH 1-2.

3. The relative expression level of SSH1 gene was determined by fluorescence quantitative PCR using 0-4cm young ear cDNA of GC-SSH1-1, GC-SSH1-2 and mutant SSH1 as templates (rice ubiquitin gene was used as reference gene, see specifically primer Ubi used in example 1). The primer for identifying the relative expression amount of SSH1 gene was the primer P3 used in example 1 above.

The relative expression level of SSH1 gene in mutant SSH1 was defined as 1, T₁The relative expression amounts of SSH1 gene in 2 lines of the generation-complementing strain are shown in FIG. 5, H. The results show that T₁The expression level in the two lines of the generation-complementary strain is obviously improved, and is 14.8 times and 36.1 times of that of the SSH1 gene in the mutant SSH1 respectively.

Fifth, obtaining transgenic over-expression plant

1. And (3) transforming the recombinant plasmid OE-SSH1 obtained in the step two into agrobacterium EHA105 by using a freeze-thaw method, transforming a transformed agrobacterium into a mutant SSH1 by using a method of infecting rice calluses with agrobacterium, performing 3 rounds of screening by using an NB culture medium containing hygromycin (the concentrations of the hygromycin used in each round of screening are 15mg/L, 25mg/L and 35mg/L), and performing pre-differentiation, differentiation and rooting to obtain a transgenic plant.

2. After step 1 is completed, extracting total DNA of transgenic plants, and performing PCR identification by using hygromycin resistance genes carried by a pCAMBIA1301 vector (the size of a target band of a positive plant is 1,254bp), wherein the primers are as follows:

Hpt_F:5’-TGCATCATCGAAATTGCCGT-3’

Hpt_R:5’-AAACGACGGCCAGTGAATTC-3’

through identification, 17 positive T strains are obtained₀The generation SSH1 gene overexpression plant. Will T₀The seed produced by selfing of the over-expressed plant of the generation SSH1 gene is named as T₁Over-expressing seeds by T₁The rice plant grown by the over-expression seed is named as T₁The generation SSH1 gene overexpression plant. Two over-expression strains were randomly selected and named OE-SSH1-1 and OE-SSH 1-2.

3. The relative expression level of the SSH1 gene was determined by fluorescence quantitative PCR using 0-4cm young ear cDNA of OE-SSH1-1, OE-SSH1-2 and mutant SSH1 as templates (rice ubiquitin gene was used as reference gene, see specifically primer Ubi used in example 1). The primer for identifying the relative expression amount of SSH1 gene was the primer P3 used in example 1 above.

The relative expression level of SSH1 gene in mutant SSH1 was defined as 1, T₁The relative expression amounts of SSH1 gene in 2 lines of the generation-overexpressed strain are shown in FIG. 5, H. The results show that T₁The expression level of the two strains of the generation over-expression strain is obviously improved and is respectively 2.2 times and 2.9 times of that of the SSH1 gene in the mutant SSH 1.

Sixth, obtaining transgenic silent strains

1. And (3) transforming the recombinant plasmid RNAi-SSH1 obtained in the third step into agrobacterium EHA105 by using a freeze-thaw method, transforming the transformed agrobacterium into wild YIL100 by using a method for infecting rice calluses by using the agrobacterium, performing 3 rounds of screening by using an NB culture medium containing hygromycin (the concentrations of the hygromycin used in each round of screening are 15mg/L, 25mg/L and 35mg/L respectively), and performing pre-differentiation, differentiation and rooting to obtain a transgenic plant.

2. After completing the step 1, extracting the total DNA of the transgenic plants, and performing PCR identification on the hygromycin resistance gene carried by the pTCK303/JL1460 vector by using the following primers (the target band size of the positive plants is 1,254 bp):

Hpt_F:5’-TGCATCATCGAAATTGCCGT-3’；

Hpt_R:5’-AAACGACGGCCAGTGAATTC-3’。

8 positive T strains are obtained through identification₀Generation SSH1 gene-silenced strain. Will T₀The seed produced by selfing of the generation SSH1 gene complementation strain is named as T₁Generation silenced seed consisting of T₁The rice plant grown by the generation silent seed is named as T₁Generation SSH1 gene-silenced strain. Two silencing strains were randomly selected and named RNAi-SSH1-1, RNAi-SSH 1-2.

3. The relative expression level of SSH1 gene was determined by fluorescence quantitative PCR using 0-4cm young ear cDNA of RNAi-SSH1-1, RNAi-SSH1-2 and mutant SSH1 as templates (rice ubiquitin gene was used as internal reference gene, see specifically primer Ubi used in example 1). The primer for identifying the SSH1 gene was the primer P3 used in example 1 above.

T₁The relative expression of SSH1 gene in 2 lines of the generation-silenced strain is shown in FIG. 5H (where WT is indica rice line YIL 100). The results showed that, when the relative expression level of SSH1 gene in mutant SSH1 was taken as 1, the relative expression level of SSH1 gene in indica rice line YIL100 was 13.6, and T was₁The expression levels in the two silencing lines of the generation strain were down-regulated to different degrees, respectively 1.0 (0.074 times of SSH1 gene in YIL100) and 11.39 (0.84 times of SSH1 gene in YIL 100).

Seventhly, obtaining of empty vector plants

1. Replacing the recombinant plasmid GC-SSH1 with the plasmid pCAMBIA1300, and operating according to the fourth step to obtain T₁The empty vector plant (pCAMBIA1300) was transferred.

2. Replacing the recombinant plasmid OE-SSH1 with the plasmid pCAMBIA1301, and performing the operation according to the fifth stepTo obtain T₁Empty vector plants (pCAMBIA1301) were transferred.

3. Replacing recombinant plasmid RNAi-SSH1 with plasmid pTCK303/JL1460, and performing the operation according to the sixth step to obtain T₁Empty vector plants (pTCK303/JL1460) were transferred.

Eight, phenotypic assay

And (3) the plant to be detected: indica rice line YIL100, mutant ssh1, T₁Generation silencing strains (RNAi-SSH1-1 and RNAi-SSH1-2) and T₁Generation complementary strains (GC-SSH1-1, GC-SSH1-2) and T₁Generation overexpression strains (OE-SSH1-1 and OE-SSH1-2) and T₁Substitute empty carrier plant (pCAMBIA1300), T₁Transfer empty vector plants (pCAMBIA1301) and T₁Empty vector plants (pTCK303/JL1460) were transferred.

Seeds of the plants to be tested are respectively sown in the field, 10 plants are randomly selected from each plant line, and the experiment is repeated for three times. The plant is investigated for the seed setting and seed length after maturation.

The experimental results are shown in figure 5(A is main stem ear, E is kernel morphology, F is shattering rate, G is kernel length, wherein WT is indica rice line YIL 100).

The results show that the phenotype of the complementation plants and the overexpression plants is basically consistent with that of the YIL100, the natural falling performance is extremely strong, and the grains are shortened. The silent strain is consistent with the mutant ssh1, the falling seed performance is reduced, and the seed is lengthened. The phenotype of the transgenic empty vector plant (pCAMBIA1300) and the transgenic empty vector plant (pCAMBIA1301) has no significant difference with the mutant ssh 1. The phenotype of the empty vector transfer plant (pTCK303/JL1460) has no significant difference with that of the wild-type YIL 100.

Sequence listing

<110> university of agriculture in China

<120> protein related to rice grain dropping property and grain type, and coding gene and application thereof

<160>4

<170>PatentIn version 3.5

<210>1

<211>436

<212>DNA

<213> Artificial sequence

<220>

<230>

<400>1

Met Val Leu Asp Leu Asn Val Glu Ser Pro Gly Gly Ser Ala Ala

1 5 10 15

Thr Ser Ser Ser Ser Thr Pro Pro Pro Pro Pro Asp Gly Gly Gly

20 25 30

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

35 40 45

Asp Gly Cys Ser Leu Pro Val MET Thr Arg Gln Leu Phe Pro Ser

50 55 60

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

65 70 75

Pro Pro Thr MET Pro Thr Pro Ala Ala Ala Gly Glu Gly Pro Trp

80 85 90

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

95 100 105

Pro Ala Gly Gly Lys Lys Ser Arg Arg Gly Pro Arg Ser Arg Ser

110 115 120

Ser Gln Tyr Arg Gly Val Thr Phe Tyr Arg Arg Thr Gly Arg Trp

125 130 135

Glu Ser His Ile Trp Asp Cys Gly Lys Gln Val Tyr Leu Gly Gly

140 145 150

Phe Asp Thr Ala His Ala Ala Ala Arg Ala Tyr Asp Arg Ala Ala

155 160 165

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

170 175 180

Asp Tyr Glu Asp Asp Leu Lys Gln MET Arg Asn Trp Thr Lys Glu

185 190 195

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

200 205 210

Gly Ser Ser Lys Tyr Arg Gly Val Thr Leu His Lys Cys Gly Arg

215 220 225

Trp Glu Ala Arg MET Gly Gln Leu Leu Gly Lys Lys Tyr Ile Tyr

230 235 240

Leu Gly Leu Phe Asp Ser Glu Ile Glu Ala Ala Arg Ala Tyr Asp

245 250 255

Arg Ala Ala Ile Arg Phe Asn Gly Arg Glu Ala Val Thr Asn Phe

260 265 270

Asp Pro Ser Ser Tyr Asp Gly Asp Val Leu Pro Glu Thr Asp Asn

275 280 285

Glu Val Val Asp Gly Asp Ile Ile Asp Leu Asn Leu Arg Ile Ser

290 295 300

Gln Pro Asn Val His Glu Leu Lys Ser Asp Gly Thr Leu Thr Gly

305 310 315

Phe Gln Leu Asn Cys Asp Ser Pro Glu Ala Ser Ser Ser Val Val

320 325 330

Thr Gln Pro Ile Ser Pro Gln Trp Pro Val Leu Pro Gln Gly Thr

335 340 345

Ser MET Ser Gln His Pro His Leu Tyr Ala Ser Pro Cys Pro Gly

350 355 360

Phe Phe Val Asn Leu Arg Glu Val Pro MET Glu Lys Arg Pro Glu

365 370 375

Leu Gly Pro Gln Ser Phe Pro Thr Ser Trp Ser Trp Gln MET Gln

380 385 390

Gly Ser Pro Leu Pro Leu Leu Pro Thr Ala Ala Ser Ser Gly Phe

395 400 405

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

410 415 420

Pro His Pro Phe Pro Gly His His Gln Phe Tyr Phe Pro Pro Thr

425 430 435

Thr ***

436

<210>2

<211>7206

<212>DNA

<213> Artificial sequence

<220>

<230>

<400>2

GGCTCCAAAG TATAAAACAC ACAAATTCTC TCAAATTTTT AAAAATGTTC AAATTGTGAG 60

TATATACCTA GGAATATAAA TTTGATTCAT CCAAATACCT AACAACAAAA CAACTCAAAC 120

TCAACTTTAT TTCACAATTC ATTATTACAT ACCTAACTAA TTATATGTTT GGATGTTATA 180

TATCTAGAAC CAAAATCTAA CAAAAAAAAG TTCAAACTTG TTAGTAAAGA AGTTAATGTT 240

CATATCTAAA CATTTAAAAA AAATTTCATA CTCATTCAAC ATACACAGTG AAAACAACAT 300

ATCCGTTTCC TTGTTTGTGT AACAAGTGTT TGGTACTTCC TGCCCATTAC TGCACGCTGG 360

TGGTGGACAG TAGCAGCACA GGAGAAGACA GCCATGGGGA TGGGGCGGCC AATGGGAAAC 420

TATACCGGTG GATATTTTGC CGACAGGAAA TGCTGCACGC ACTAACCAAT GTTGCTCACA 480

AAGTTTTTTT TTAAAATTTT TAATTTCTCC CTCGCATCAT GTGGTCCCTG CAGATACATC 540

ATGTCATAAA AGTTTTTCTT AACAGTGAAG TATTATTATT ATTATTGTTT GCTCCTTTGA 600

TATTTATACTTGATTATGAA ATCAAGTTTA TTGTATTATG TAGAATATGC AGTGCCATGC 660

TTGGGTGTCT CTCTCGTCCA GAAGCTGAGT TAGACTTCAT TTTTTTAAAA GATTGTACAT 720

TAATTTAACC ACAACCTAGA AAGTGCTTGT CTTGTGCATA TAAAAACATG TATGTTATAT 780

TATTTTTTTG TGAGGAATAT TATGGAATGG TGGAATATAT TTATCTCCTT TTAAAGAAGC 840

ACCTTGTTTC TATATTTCAT ATCCATACCT TTAAAAACAT CACGTTGTAT TTTGCTCCTT 900

CGTTTGCCCG TTAGCTTATG AGTCAAAGCA AAATTTGAAT TTTTAGACTT AATTCTAGAG 960

TTAATTTTGA GGCATTTTTT TTATTATAGT TTAGTTTTCA GCATTGGTTT TTAAAATTGC 1020

CAAGAGCACA TATATACAAA TTTTATACAT AAATCAATTT TTGTTGCTTC ATTCTATTTT 1080

ACAACTTAAT GGCAGAGCAA ACGATGAAAA TAATTTATAT CGTGTGCTCT AGAAATGTTG 1140

TTTTCACTAT ATCATGGATC AGTTTAGGTA GCCGTGTAGA TATCCTTACT CCGTAATAAA 1200

ACGAGCCAAC AATGCTAGCT TAGAGCCGTT AATTATTGTT ACAAGTAAAT GCAACCTTAA 1260

TCTATATATG CTTATCTGAC TTCAAAAGAA GTACTCCAAC CATGTAACCA AAATCCAACC 1320

ATGTTATAAG TCGATTTAAT TATTCTCTAG CTACTTACTA AAGTTTGATA TACTCTTATA 1380

TAGAGAAAAT TAGCAACATC TACAATACAA AATTAGTTTT ATTCTAACAT TCAACACATT 1440

TTGATAATAC GTTTTAAACA TTAGTATATA ATTCTAAAAA ATAAATTAAA GCAATGTCGA 1500

GTGGAAAATA CAACTTTTAT CACTGTGATG CAATCCTATA TACGACTACC TTTATATACT 1560

ACTCCATTTT GGGTTACAAG ACGTTTTTAC TTTGGTCAAA ATCAAACTCT TTCAAATTTA 1620

ACTAAATTTA TAGATACATA TAGTAATATT TATAATACTA AATTAGTTTG ATTCAATCAA 1680

TAATTGAATA TATATTCATA ATAAATTTGT CTTGGGTTGA AAATGTATTA TTTTTTTTAC 1740

AAACTTGGTT AAACCTAAAG CAGTTTAACT TTGATAAAAG GTCAAAACGT CTTATAATCT 1800

AGATTAGAGG GAATAGTACC AGTAGTCGGC AAACACATTG CAACTATTGG GCCAGAGTCA 1860

AGTCAAACTA AGCATAGCAC ACTCCTCCTT AATACACCCT ACATACACAA AACCATACTA 1920

CTACTACTAG TAGGAGTCTC CACCGTTCTG AAAAAAGAAA AAGAAAAAAA AAACCCTCCA 1980

TCTCCCGCCC AGTGCCGCCG CGCCACGTCA CCCCTCCCCC GCCGCGCGTG TGTGTACGCG 2040

TAGCCTTTCC GGCGGAGACG CAGCACAAAA ATGGTTTTAC CTGCGATACC CTTCGTCTCC 2100

CTCACGTCCC ATCCATCCCT CCCCCCCCTT TTCCCTCCTC CTCCACCTCC ACTGCCATGG 2160

CCGCCTCCGC CGCTCCTCTC TCCCGCGTGG TAGCCGCCGC CGGTTGCCGC CGCTAGGGGA 2220

GCGCGACGGG GGAGCGGTAG GGGCGACCCA GCTCGCTCTG TCGCCCCGCG GCTGGTGGGG 2280

GTGCGCGCGG GGGAGAGCGG TTGGTTAGTA TGGTGCTGGA TCTCAATGTG GAGTCGCCGG 2340

GTGGGTCGGC GGCGACGTCG AGCTCGTCCA CGCCGCCGCC GCCGCCCGAC GGTGGCGGCG 2400

GGGGGTACTT CCGGTTCGAC CTGCTCGGCG GGAGCCCCGA CGAGGACGGG TGCTCCCTGC 2460

CTGTCATGAC GCGCCAGCTC TTCCCTTCGC CGTCTGCGGT GGTGGCGCTG GCGGGGGACG 2520

GGTCGTCGAC GCCACCGCCG ACGATGCCGA CGCCGGCGGC GGCTGGGGAG GGGCCGTGGC 2580

CGCGCCGCGC GGCGGATCTC GGGGTGGCGC AGAGCCAGAG GTCCCCCGCC GGCGGGAAGA 2640

AGAGCCGCCG CGGCCCGAGG TCTCGGAGCT CCCAGTACAG GGGCGTCACC TTCTACAGGA 2700

GGACCGGGCG ATGGGAGTCG CACATCTGGT TAGCTTCGCC TTGCTCGATT TCCCTCACGC 2760

TGACTTCCTC TGCTTGACTC TGCTTGATTG GAGTAGTAATCTCTTCGTTT GTCTCGTCCA 2820

ATTTTGGCAG GGACTGCGGG AAGCAGGTGT ACCTGGGTGA GCTCCTATTT CTAGTCTCCC 2880

AAGCTAAATT CGCTCGCATG ACTGCTAAAT TTGGCTCTCT ACTTAGCTTG ATTCCGATTT 2940

GTTTGCTCCT GCCTTGTTTT TTTTTTTTCT GTTTTCAGGT GGTTTCGATA CAGCTCATGC 3000

CGCAGCGAGG TTACTATAGA TGGTCACCAA TTGTATCTTA ATTGTTGCTC TGGTTTGCCC 3060

TGTGTGCTGA GATTGTAGAA CCCCTTGTGC AGGGCCTATG ATCGCGCGGC GATCAAGTTC 3120

AGAGGCCTCG ACGCGGATAT CAACTTTAAT CTGAATGACT ATGAGGACGA CTTGAAGCAG 3180

GTTGTTTGGA ATATAGTTCA ATTGTCATGT GTGATTCTTT AGGTTCTAAG GGGAGGTTCA 3240

GTCACCGGCC TGTGATGGTT TATGATGGAT TGGTTCGTGC TGTGTTGTTA GATGCGCAAT 3300

TGGACCAAGG AGGAGTTTGT GCACATACTT CGGCGCCAAA GCACAGGATT TGCAAGGGGG 3360

AGCTCAAAGT ACCGGGGTGT GACACTGCAC AAGTGTGGCC GGTGGGAAGC TCGGATGGGC 3420

CAGCTGCTCG GCAAGAAGTA AGATCCCTGA AACATTGATT GTTCTTGCTA GTAACTTACA 3480

ATACAATACA TTTTAGATTA GTTCAGGACC ATTTTTCATG CCATGAACGA AAACTGTGTC 3540

AAGTTTGCTT TTCAAAAAGG GAAAGAAAAA AAATGTGTCA CTACCAAGCT GGAGAATGGT 3600

AGGCTACCCC TGGAGTCCAC ATCCACCTGA AGATTTGGCA CCTTGATGTT GACTCTTCAC 3660

ACACTGGCCA TTTACTATTG GAACACCACA GGTGTTCAGT GATTATAGAT TGCTAGCTTC 3720

TTTAGGAAAG AACTAAGTGA ATCGGAATGG CTCAACGGAA TTAATTGAGG GGAAATGTTT 3780

TTGTTTCCTT GGGGATTGTG TTTTGCGTCC TAAGTCAGTG ACAGGTGAAA AGCTTCATTT 3840

CAATGCTCTT GGTTGTTTTT GCCACTAAGA ATGTTCTGCT TATTTACGTT GAATTTTTGA 3900

TTGGCTGTTC GTATCATAGT TGTGCATTTC CTATATATTA CAATCTTTGT GAACTTTAGA 3960

TAATTCTGGC ATGAGCTCTC TCTGTGGTAT TGTTCTATTA ACAGAAGTAT CATTCTGTCA 4020

ATTCCCCACT GATAAGTGAT CTTTTGTGCA GGTACATCTA TCTAGGATTG TTTGACAGTG 4080

AAATTGAGGC TGCAAGGTTC AGCTAGTAGT ACTTTGCTCT TCTCAGTTTG ATATGTGCTT 4140

GCTATTCCTA TACTTCTCTC ACTTTCATCT ACTCTTCAAC TTGTAGAGCA TATGACCGGG 4200

CAGCTATCCG CTTCAATGGA AGGGAAGCTG TTACTAATTT TGATCCTAGT TCTTATGATG 4260

GAGATGTTCT ACCTGAAACC GACAATGAAG GTATTTCTTT TGTACTATGT ATATTGTGCT 4320

TTATCCACTA GAACGACAAA TCCAAATTTT ATATTAACAA AAGCACACTG GGGATTGTCT 4380

TAGCAGTGGT TGATGGAGAC ATCATTGACT TAAATCTGAG AATTTCACAG CCTAACGTTC 4440

ATGAGCTGAA AAGTGATGGT ACCCTAACTG GGTTCCAGTT GAATTGTGAT TCTCCTGAAG 4500

CTTCAAGTTC TGTTGTTACT CAGGTAAATA TGCTACAATA TTTGGTGTTT GTAACCGCTA 4560

TACCATTTGA GATTTGACCG TCTTCAGATT CTTTTGATCA GGTGGTTAGT CCTTTTTCAT 4620

AACATTATAC TATCACTTTC AAAATAATTA GTACACTAAT TGTATGTGTC ACTAGAATTG 4680

TTGTATGAAA CCATCTGGGA ACCATCTTAT GCAGCTTATT GACAAATGAC AACTCACTGT 4740

TGATGTATTC CCAAATCAAG AAAAGCCAAT CTTTTTATTC AATTTCTCCA GTTCATTGAA 4800

TTGAAAAAGA AATTGCCGAA CAACTACCAC CTGAAGAGAC TGGCTAGATA TTTTGTTTAT 4860

CTGTGTATTT TTATAACCTA TAACAGTTCC AAAGCCACAA AGCTCTTTTA CCTTGATTCT 4920

GATAACGTTA ACTGAGGTAT CATTTTGTGA TTGTTTTCTG TAGCCAATAA GTCCTCAGTG 4980

GCCTGTGCTT CCTCAGGGCA CATCGATGTC CCAGCATCCA CATTTATATG CATCTCCTTG 5040

TCCGGGCTTC TTTGTGAACC TCAGGGTATA TCTATCATCA AAACTTATAT GCAATTTGAA 5100

AAGACAGTTA AACTTGTTTC AAGTTAACAT TCGATTGCTA ATTAGCTACT ACTTCCATGC 5160

TCCTATTTCT TGAGATGACG AACCATATGT TTGTTCCTTT AATTACATAA TGGTAGCCGT 5220

AGATACTGTT TGCCTTGTGA ACTGTCAGAT ATAGCAGGAA GTAAATCTGT CAGCTTGCAA 5280

CTTTCCATGA ACCACAGCTG TCCAACTTTA TACAGTGGAT GATGACTTGC ATTTGATACA 5340

TCCTAGTACT TTCTTGTTGC ACTCTGCAGC TTTAGCTCTG AAGATGCTCT ATATTTCCAT 5400

GAATGCTCAC GTTAGCTCTG AAATTTCGAA CTTTCACTCC ATAACATGCT CATTTAGGCT 5460

GTGTTTAGAT CCAAACTTCA GTTCTTTTCC ATCACATCAA CCTGTCATAC ACACACAACT 5520

TTTCAGTCAC ATCATCTCTA ATTTCAACCA AAATCCAAAC TTTGCCATCA ACTAAACACA 5580

GCCTTATTCA TTGTTCCACC TTTTATTTCT GCCATTGCTA TGGTCCAATC TTGATACCTC 5640

AGAGAGCTGC ATGATGTGTA GAAGTCCTAA CTTAGGTCCG TTGGTTAGAA TCATCCATTC 5700

ATGTTTAGGA AAAGTGATTA ACAACGTGTG CTGCATTAAC CATTAATGTG GTCAACTCTG 5760

ATGCTTGTGC TATGGTCACT GCAGGAAGTA CCTATGGAGA AAAGACCTGA GTTGGGTCCC 5820

CAGTCGTTCC CTACTTCGTG GTCATGGCAA ATGCAGGGCT CCCCTTTGCC ATTACTCCCT 5880

ACTGCAGCAT CATCAGGATT CTCTACGGGC ACCGTCGCCG ACGCCGCCCG CGCGCCTTCC 5940

TCCCGCCCCC ATCCATTTCC CGGCCACCAC CAGTTCTACT TCCCCCCGAC CACCTGACTG 6000

CCACCTATTC TGGTGGAGGC GACGCCTCAC CGTGCATCCA CCGCCGCTTG CCGATAACAT 6060

TCGTCGTTTG TCCAGAGAGG GACCTTCTCC ATATGATATC CCTCTCTATG TTCCACCTGG 6120

TTATGATCAG AATTCTCACG CTCATGATTC TTTTATCTTC ATTTTTGAGT GCGAAACCAC 6180

CAATCGTGAC CGTGCTACCA GGTAAACACC CTTGTACCTC TTTGACTCTA CAATAAAATT 6240

ATTGTAAGAT ATTCTGGCTA AAAGTGATTG GCCCTCCCTA CTGTATTTTT ACAGAGTTGT 6300

AATTGAACAG TGGGCACTAA AGCTGGGGCC CACTCGCACC AGTGTCTAGT GGAGCGAAGC 6360

TTTGCCCTTT TCTTGCTATC CAGCAACTAT TCCTCCGCCT GCCTCCATTG CTGACAGTGG 6420

AGATAGCTTC CCAGTGGTCA CTGTTCCACT GTACTCTGTC ATAGTTTCTG ATCACCCCTA 6480

TTGTGCCTTG TCTCTTTCAC CTCCTCCCTC ACTTGTGCTG CCTCTGGCCA CTAGAGTGGC 6540

GCCCTGCACT GTTGCCATGC ATGGCTGCCA CTTCGAGAGT GGAGATGGGG GTTGGCCTTT 6600

GGGTGATTTC TGTGTGCCAT CTTTTGGCTC AATGTCGTTT TGGGACTGGT GCATGTACTG 6660

GCAATAGGTG AGATGAGATA TGAAATGTCC TCTGCTGCTT TTTCTATCTA AGGAGTAGCT 6720

TAGCTATCAC CAGCTGCCTT TCTCTTCTCA AATAAGGTTG TTAAAAGGGG GGGTCTTAAA 6780

GCTACCCCAA CTGCTGCCAG GGGCCCTCTT CAATGCCCTC CCAGCTGCCG TTTCTTTCTT 6840

TATTGGTTTG CTTCTCTGCT GCTTCTTTTG CCTGGTTGGA TGGATGTTTG TTTGTTCGTA 6900

TGAATTGGGG GAGCTACTAG TATCATTCAT GAGTAGAGGC AGGCAGGGCA GCGCAGGCTG 6960

CTGGCTGGCC AGGCACTGAT GTGCCAACAG CTATTTCCAG TGAAACGGCT GCCATTTTTC 7020

TTTGCTGGGA TTGTGATAGT GGTAGGCTGG TAGTACTAGC TAGCAGTGTA TTGGGGGAAA 7080

TACTGGTGGT CCATTTGCCT GTTAGCCAAT CTGATGCTTG CCTTTCCATC CGCTGGTGAT 7140

GGTCCATTTG CTTTGGCATC TTGCTTGCCT GTTAGTACTA CAAAAGTTGC ATACATATGC 7200

TATTGA 7206

<210>3

<211>7206

<212>DNA

<213> Artificial sequence

<220>

<230>

<400>3

GGCTCCAAAG TATAAAACAC ACAAATTCTC TCAAATTTTT AAAAATGTTC AAATTGTGAG 60

TATATACCTA GGAATATAAA TTTGATTCAT CCAAATACCT AACAACAAAA CAACTCAAAC 120

TCAACTTTAT TTCACAATTC ATTATTACAT ACCTAACTAA TTATATGTTT GGATGTTATA 180

TATCTAGAAC CAAAATCTAA CAAAAAAAAG TTCAAACTTG TTAGTAAAGA AGTTAATGTT 240

CATATCTAAA CATTTAAAAA AAATTTCATA CTCATTCAAC ATACACAGTG AAAACAACAT 300

ATCCGTTTCC TTGTTTGTGT AACAAGTGTT TGGTACTTCC TGCCCATTAC TGCACGCTGG 360

TGGTGGACAG TAGCAGCACA GGAGAAGACA GCCATGGGGA TGGGGCGGCC AATGGGAAAC 420

TATACCGGTG GATATTTTGC CGACAGGAAA TGCTGCACGC ACTAACCAAT GTTGCTCACA 480

AAGTTTTTTT TTAAAATTTT TAATTTCTCC CTCGCATCAT GTGGTCCCTG CAGATACATC 540

ATGTCATAAA AGTTTTTCTT AACAGTGAAG TATTATTATT ATTATTGTTT GCTCCTTTGA 600

TATTTATACT TGATTATGAA ATCAAGTTTA TTGTATTATG TAGAATATGC AGTGCCATGC 660

TTGGGTGTCT CTCTCGTCCA GAAGCTGAGT TAGACTTCAT TTTTTTAAAA GATTGTACAT 720

TAATTTAACC ACAACCTAGA AAGTGCTTGT CTTGTGCATA TAAAAACATG TATGTTATAT 780

TATTTTTTTG TGAGGAATAT TATGGAATGG TGGAATATAT TTATCTCCTT TTAAAGAAGC 840

ACCTTGTTTC TATATTTCAT ATCCATACCT TTAAAAACAT CACGTTGTAT TTTGCTCCTT 900

CGTTTGCCCG TTAGCTTATG AGTCAAAGCA AAATTTGAAT TTTTAGACTT AATTCTAGAG 960

TTAATTTTGA GGCATTTTTT TTATTATAGT TTAGTTTTCA GCATTGGTTT TTAAAATTGC 1020

CAAGAGCACA TATATACAAA TTTTATACAT AAATCAATTT TTGTTGCTTC ATTCTATTTT 1080

ACAACTTAAT GGCAGAGCAA ACGATGAAAA TAATTTATAT CGTGTGCTCT AGAAATGTTG 1140

TTTTCACTAT ATCATGGATC AGTTTAGGTA GCCGTGTAGA TATCCTTACT CCGTAATAAA 1200

ACGAGCCAAC AATGCTAGCT TAGAGCCGTT AATTATTGTT ACAAGTAAAT GCAACCTTAA 1260

TCTATATATG CTTATCTGAC TTCAAAAGAA GTACTCCAAC CATGTAACCA AAATCCAACC 1320

ATGTTATAAG TCGATTTAAT TATTCTCTAG CTACTTACTA AAGTTTGATA TACTCTTATA 1380

TAGAGAAAAT TAGCAACATC TACAATACAA AATTAGTTTT ATTCTAACAT TCAACACATT 1440

TTGATAATAC GTTTTAAACA TTAGTATATA ATTCTAAAAA ATAAATTAAA GCAATGTCGA 1500

GTGGAAAATA CAACTTTTAT CACTGTGATG CAATCCTATA TACGACTACC TTTATATACT 1560

ACTCCATTTT GGGTTACAAG ACGTTTTTAC TTTGGTCAAA ATCAAACTCT TTCAAATTTA 1620

ACTAAATTTA TAGATACATA TAGTAATATT TATAATACTA AATTAGTTTG ATTCAATCAA 1680

TAATTGAATA TATATTCATA ATAAATTTGT CTTGGGTTGA AAATGTATTA TTTTTTTTAC 1740

AAACTTGGTT AAACCTAAAG CAGTTTAACT TTGATAAAAG GTCAAAACGT CTTATAATCT 1800

AGATTAGAGG GAATAGTACC AGTAGTCGGC AAACACATTG CAACTATTGG GCCAGAGTCA 1860

AGTCAAACTA AGCATAGCAC ACTCCTCCTT AATACACCCT ACATACACAA AACCATACTA 1920

CTACTACTAG TAGGAGTCTC CACCGTTCTG AAAAAAGAAA AAGAAAAAAA AAACCCTCCA 1980

TCTCCCGCCC AGTGCCGCCG CGCCACGTCA CCCCTCCCCC GCCGCGCGTG TGTGTACGCG 2040

TAGCCTTTCC GGCGGAGACG CAGCACAAAA ATGGTTTTAC CTGCGATACC CTTCGTCTCC 2100

CTCACGTCCC ATCCATCCCT CCCCCCCCTT TTCCCTCCTC CTCCACCTCC ACTGCCATGG 2160

CCGCCTCCGC CGCTCCTCTC TCCCGCGTGG TAGCCGCCGC CGGTTGCCGC CGCTAGGGGA 2220

GCGCGACGGG GGAGCGGTAG GGGCGACCCA GCTCGCTCTG TCGCCCCGCG GCTGGTGGGG 2280

GTGCGCGCGG GGGAGAGCGG TTGGTTAGTA TGGTGCTGGA TCTCAATGTG GAGTCGCCGG 2340

GTGGGTCGGC GGCGACGTCG AGCTCGTCCA CGCCGCCGCC GCCGCCCGAC GGTGGCGGCG 2400

GGGGGTACTT CCGGTTCGAC CTGCTCGGCG GGAGCCCCGA CGAGGACGGG TGCTCCCTGC 2460

CTGTCATGAC GCGCCAGCTC TTCCCTTCGC CGTCTGCGGT GGTGGCGCTG GCGGGGGACG 2520

GGTCGTCGAC GCCACCGCCG ACGATGCCGA CGCCGGCGGC GGCTGGGGAG GGGCCGTGGC 2580

CGCGCCGCGC GGCGGATCTC GGGGTGGCGC AGAGCCAGAG GTCCCCCGCC GGCGGGAAGA 2640

AGAGCCGCCG CGGCCCGAGG TCTCGGAGCT CCCAGTACAG GGGCGTCACC TTCTACAGGA 2700

GGACCGGGCG ATGGGAGTCG CACATCTGGT TAGCTTCGCC TTGCTCGATT TCCCTCACGC 2760

TGACTTCCTC TGCTTGACTC TGCTTGATTG GAGTAGTAAT CTCTTCGTTT GTCTCGTCCA 2820

ATTTTGGCAG GGACTGCGGG AAGCAGGTGT ACCTGGGTGA GCTCCTATTT CTAGTCTCCC 2880

AAGCTAAATT CGCTCGCATG ACTGCTAAAT TTGGCTCTCT ACTTAGCTTG ATTCCGATTT 2940

GTTTGCTCCT GCCTTGTTTT TTTTTTTTCT GTTTTCAGGTGGTTTCGATA CAGCTCATGC 3000

CGCAGCGAGG TTACTATAGA TGGTCACCAA TTGTATCTTA ATTGTTGCTC TGGTTTGCCC 3060

TGTGTGCTGA GATTGTAGAA CCCCTTGTGC AGGGCCTATG ATCGCGCGGC GATCAAGTTC 3120

AGAGGCCTCG ACGCGGATAT CAACTTTAAT CTGAATGACT ATGAGGACGA CTTGAAGCAG 3180

GTTGTTTGGA ATATAGTTCA ATTGTCATGT GTGATTCTTT AGGTTCTAAG GGGAGGTTCA 3240

GTCACCGGCC TGTGATGGTT TATGATGGAT TGGTTCGTGC TGTGTTGTTA GATGCGCAAT 3300

TGGACCAAGG AGGAGTTTGT GCACATACTT CGGCGCCAAA GCACAGGATT TGCAAGGGGG 3360

AGCTCAAAGT ACCGGGGTGT GACACTGCAC AAGTGTGGCC GGTGGGAAGC TCGGATGGGC 3420

CAGCTGCTCG GCAAGAAGTA AGATCCCTGA AACATTGATT GTTCTTGCTA GTAACTTACA 3480

ATACAATACA TTTTAGATTA GTTCAGGACC ATTTTTCATG CCATGAACGA AAACTGTGTC 3540

AAGTTTGCTT TTCAAAAAGG GAAAGAAAAA AAATGTGTCA CTACCAAGCT GGAGAATGGT 3600

AGGCTACCCC TGGAGTCCAC ATCCACCTGA AGATTTGGCA CCTTGATGTT GACTCTTCAC 3660

ACACTGGCCA TTTACTATTG GAACACCACA GGTGTTCAGT GATTATAGAT TGCTAGCTTC 3720

TTTAGGAAAG AACTAAGTGA ATCGGAATGG CTCAACGGAA TTAATTGAGG GGAAATGTTT 3780

TTGTTTCCTT GGGGATTGTG TTTTGCGTCC TAAGTCAGTG ACAGGTGAAA AGCTTCATTT 3840

CAATGCTCTT GGTTGTTTTT GCCACTAAGA ATGTTCTGCT TATTTACGTT GAATTTTTGA 3900

TTGGCTGTTC GTATCATAGT TGTGCATTTC CTATATATTA CAATCTTTGT GAACTTTAGA 3960

TAATTCTGGC ATGAGCTCTC TCTGTGGTAT TGTTCTATTA ACAGAAGTAT CATTCTGTCA 4020

ATTCCCCACT GATAAGTGAT CTTTTGTGCA GGTACATCTA TCTAGGATTG TTTGACAGTG 4080

AAATTGAGGC TGCAAGGTTC AGCTAGTAGT ACTTTGCTCT TCTCAGTTTG ATATGTGCTT 4140

GCTATTCCTA TACTTCTCTC ACTTTCATCT ACTCTTCAAC TTGTAGAGCA TATGACCGGG 4200

CAGCTATCCG CTTCAATGGA AGGGAAGCTG TTACTAATTT TGATCCTAGT TCTTATGATG 4260

GAGATGTTCT ACCTGAAACC GACAATGAAG GTATTTCTTT TGTACTATGT ATATTGTGCT 4320

TTATCCACTA GAACGACAAA TCCAAATTTT ATATTAACAA AAGCACACTG GGGATTGTCT 4380

TAGCAGTGGT TGATGGAGAC ATCATTGACT TAAATCTGAG AATTTCACAG CCTAACGTTC 4440

ATGAGCTGAA AAGTGATGGT ACCCTAACTG GGTTCCAGTT GAATTGTGAT TCTCCTGAAG 4500

CTTCAAGTTC TGTTGTTACT CAGGTAAATA TGCTACAATA TTTGGTGTTT GTAACCGCTA 4560

TACCATTTGA GATTTGACCG TCTTCAGATT CTTTTGATCA GGTGGTTAGT CCTTTTTCAT 4620

AACATTATAC TATCACTTTC AAAATAATTA GTACACTAAT TGTATGTGTC ACTAGAATTG 4680

TTGTATGAAA CCATCTGGGA ACCATCTTAT GCAGCTTATT GACAAATGAC AACTCACTGT 4740

TGATGTATTC CCAAATCAAG AAAAGCCAAT CTTTTTATTC AATTTCTCCA GTTCATTGAA 4800

TTGAAAAAGA AATTGCCGAA CAACTACCAC CTGAAGAGAC TGGCTAGATA TTTTGTTTAT 4860

CTGTGTATTT TTATAACCTA TAACAGTTCC AAAGCCACAA AGCTCTTTTA CCTTGATTCT 4920

GATAACGTTA ACTGAGGTAT CATTTTGTGA TTGTTTTCTG TAGCCAATAA GTCCTCAGTG 4980

GCCTGTGCTT CCTCAGGGCA CATCGATGTC CCAGCATCCA CATTTATATG CATCTCCTTG 5040

TCCGGGCTTC TTTGTGAACC TCAGGGTATA TCTATCATCA AAACTTATAT GCAATTTGAA 5100

AAGACAGTTA AACTTGTTTC AAGTTAACAT TCGATTGCTA ATTAGCTACT ACTTCCATGC 5160

TCCTATTTCT TGAGATGACG AACCATATGT TTGTTCCTTT AATTACATAA TGGTAGCCGT 5220

AGATACTGTT TGCCTTGTGA ACTGTCAGAT ATAGCAGGAA GTAAATCTGT CAGCTTGCAA 5280

CTTTCCATGA ACCACAGCTG TCCAACTTTA TACAGTGGAT GATGACTTGC ATTTGATACA 5340

TCCTAGTACT TTCTTGTTGC ACTCTGCAGC TTTAGCTCTG AAGATGCTCT ATATTTCCAT 5400

GAATGCTCAC GTTAGCTCTG AAATTTCGAA CTTTCACTCC ATAACATGCT CATTTAGGCT 5460

GTGTTTAGAT CCAAACTTCA GTTCTTTTCC ATCACATCAA CCTGTCATAC ACACACAACT 5520

TTTCAGTCAC ATCATCTCTA ATTTCAACCA AAATCCAAAC TTTGCCATCA ACTAAACACA 5580

GCCTTATTCA TTGTTCCACC TTTTATTTCT GCCATTGCTA TGGTCCAATC TTGATACCTC 5640

AGAGAGCTGC ATGATGTGTA GAAGTCCTAA CTTAGGTCCG TTGGTTAGAA TCATCCATTC 5700

ATGTTTAGGA AAAGTGATTA ACAACGTGTG CTGCATTAAC CATTAATGTG GTCAACTCTG 5760

ATGCTTGTGC TATGGTCACT GAAGGAAGTA CCTATGGAGA AAAGACCTGA GTTGGGTCCC 5820

CAGTCGTTCC CTACTTCGTG GTCATGGCAA ATGCAGGGCT CCCCTTTGCC ATTACTCCCT 5880

ACTGCAGCAT CATCAGGATT CTCTACGGGC ACCGTCGCCG ACGCCGCCCG CGCGCCTTCC 5940

TCCCGCCCCC ATCCATTTCC CGGCCACCAC CAGTTCTACT TCCCCCCGAC CACCTGACTG 6000

CCACCTATTC TGGTGGAGGC GACGCCTCAC CGTGCATCCA CCGCCGCTTG CCGATAACAT 6060

TCGTCGTTTG TCCAGAGAGG GACCTTCTCC ATATGATATC CCTCTCTATG TTCCACCTGG 6120

TTATGATCAG AATTCTCACG CTCATGATTC TTTTATCTTC ATTTTTGAGT GCGAAACCAC 6180

CAATCGTGAC CGTGCTACCA GGTAAACACC CTTGTACCTC TTTGACTCTA CAATAAAATT 6240

ATTGTAAGAT ATTCTGGCTA AAAGTGATTG GCCCTCCCTA CTGTATTTTT ACAGAGTTGT 6300

AATTGAACAG TGGGCACTAA AGCTGGGGCC CACTCGCACC AGTGTCTAGT GGAGCGAAGC 6360

TTTGCCCTTT TCTTGCTATC CAGCAACTAT TCCTCCGCCT GCCTCCATTG CTGACAGTGG 6420

AGATAGCTTC CCAGTGGTCA CTGTTCCACT GTACTCTGTC ATAGTTTCTG ATCACCCCTA 6480

TTGTGCCTTG TCTCTTTCAC CTCCTCCCTC ACTTGTGCTG CCTCTGGCCA CTAGAGTGGC 6540

GCCCTGCACT GTTGCCATGC ATGGCTGCCA CTTCGAGAGT GGAGATGGGG GTTGGCCTTT 6600

GGGTGATTTC TGTGTGCCAT CTTTTGGCTC AATGTCGTTT TGGGACTGGT GCATGTACTG 6660

GCAATAGGTG AGATGAGATA TGAAATGTCC TCTGCTGCTT TTTCTATCTA AGGAGTAGCT 6720

TAGCTATCAC CAGCTGCCTT TCTCTTCTCA AATAAGGTTG TTAAAAGGGG GGGTCTTAAA 6780

GCTACCCCAA CTGCTGCCAG GGGCCCTCTT CAATGCCCTC CCAGCTGCCG TTTCTTTCTT 6840

TATTGGTTTG CTTCTCTGCT GCTTCTTTTG CCTGGTTGGA TGGATGTTTG TTTGTTCGTA 6900

TGAATTGGGG GAGCTACTAG TATCATTCAT GAGTAGAGGC AGGCAGGGCA GCGCAGGCTG 6960

CTGGCTGGCC AGGCACTGAT GTGCCAACAG CTATTTCCAG TGAAACGGCT GCCATTTTTC 7020

TTTGCTGGGA TTGTGATAGT GGTAGGCTGG TAGTACTAGC TAGCAGTGTA TTGGGGGAAA 7080

TACTGGTGGT CCATTTGCCT GTTAGCCAAT CTGATGCTTG CCTTTCCATC CGCTGGTGAT 7140

GGTCCATTTG CTTTGGCATC TTGCTTGCCT GTTAGTACTA CAAAAGTTGC ATACATATGC 7200

TATTGA 7206

<210>4

<211>1909

<212>DNA

<213> Artificial sequence

<220>

<230>

<400>4

ATCCCTCCCC CCCCTTTTCC CTCCTCCTCC ACCTCCACTG CCATGGCCGC CTCCGCCGCT 60

CCTCTCTCCC GCGTGGTAGC CGCCGCCGGT TGCCGCCGCT AGGGGAGCGC GACGGGGGAG 120

CGGTAGGGGC GACCCAGCTC GCTCTGTCGC CCCGCGGCTG GTGGGGGTGC GCGCGGGGGA 180

GAGCGGTTGG TTAGTATGGT GCTGGATCTC AATGTGGAGT CGCCGGGTGG GTCGGCGGCG 240

ACGTCGAGCT CGTCCACGCC GCCGCCGCCG CCCGACGGTG GCGGCGGGGG GTACTTCCGG 300

TTCGACCTGC TCGGCGGGAG CCCCGACGAG GACGGGTGCT CCCTGCCTGT CATGACGCGC 360

CAGCTCTTCC CTTCGCCGTC TGCGGTGGTG GCGCTGGCGG GGGACGGGTC GTCGACGCCA 420

CCGCCGACGA TGCCGACGCC GGCGGCGGCT GGGGAGGGGC CGTGGCCGCG CCGCGCGGCG 480

GATCTCGGGG TGGCGCAGAG CCAGAGGTCC CCCGCCGGCG GGAAGAAGAG CCGCCGCGGC 540

CCGAGGTCTC GGAGCTCCCA GTACAGGGGC GTCACCTTCT ACAGGAGGAC CGGGCGATGG 600

GAGTCGCACA TCTGGGACTG CGGGAAGCAG GTGTACCTGG GTGGTTTCGA TACAGCTCAT 660

GCCGCAGCGA GGGCCTATGA TCGCGCGGCG ATCAAGTTCA GAGGCCTCGA CGCGGATATC 720

AACTTTAATC TGAATGACTA TGAGGACGAC TTGAAGCAGA TGCGCAATTG GACCAAGGAG 780

GAGTTTGTGC ACATACTTCG GCGCCAAAGC ACAGGATTTG CAAGGGGGAG CTCAAAGTAC 840

CGGGGTGTGA CACTGCACAA GTGTGGCCGG TGGGAAGCTC GGATGGGCCA GCTGCTCGGC 900

AAGAAGTACA TCTATCTAGG ATTGTTTGAC AGTGAAATTG AGGCTGCAAG AGCATATGAC 960

CGGGCAGCTA TCCGCTTCAA TGGAAGGGAA GCTGTTACTA ATTTTGATCC TAGTTCTTAT 1020

GATGGAGATG TTCTACCTGA AACCGACAAT GAAGTGGTTG ATGGAGACAT CATTGACTTA 1080

AATCTGAGAA TTTCACAGCC TAACGTTCAT GAGCTGAAAA GTGATGGTAC CCTAACTGGG 1140

TTCCAGTTGA ATTGTGATTC TCCTGAAGCT TCAAGTTCTG TTGTTACTCA GCCAATAAGT 1200

CCTCAGTGGC CTGTGCTTCC TCAGGGCACA TCGATGTCCC AGCATCCACA TTTATATGCA 1260

TCTCCTTGTC CGGGCTTCTT TGTGAACCTC AGGGAAGTAC CTATGGAGAA AAGACCTGAG 1320

TTGGGTCCCC AGTCGTTCCC TACTTCGTGG TCATGGCAAA TGCAGGGCTC CCCTTTGCCA 1380

TTACTCCCTA CTGCAGCATC ATCAGGATTC TCTACGGGCA CCGTCGCCGA CGCCGCCCGC 1440

GCGCCTTCCT CCCGCCCCCA TCCATTTCCC GGCCACCACC AGTTCTACTT CCCCCCGACC 1500

ACCTGACTGC CACCTATTCT GGTGGAGGCG ACGCCTCACC GTGCATCCAC CGCCGCTTGC 1560

CGATAACATT CGTCGTTTGT CCAGAGAGGG ACCTTCTCCA TATGATATCC CTCTCTATGT 1620

TCCACCTGGT TATGATCAGA ATTCTCACGC TCATGATTCT TTTATCTTCA TTTTTGAGTG 1680

CGAAACCACC AATCGTGACC GTGCTACCAG GTAAACACCC TTGTACCTCT TTGACTCTAC 1740

AATAAAATTA TTGTAAGATA TTCTGGCTAA AAGTGATTGG CCCTCCCTAC TGTATTTTTA 1800

CAGAGTTGTA ATTTGAACAG TGGGCACTAA AGCTGGGGCC CACTCGCACC AGTGTCTAGT 1860

GGAGCGAAGC TTTGCCCTTT TCTTGCTATC CAGCAACTAT TCCTCCGCC 1909

Claims

1. The application of a protein or a coding gene thereof in regulating and controlling the seed shattering property and/or the seed type of a plant; the amino acid sequence of the protein is shown as a sequence 1 in a sequence table.

2. Use according to claim 1, characterized in that: the encoding gene is a DNA molecule as described in any one of (b1) to (b3) below:

(b3) a DNA molecule shown as 2310-5997 th nucleotide of a sequence 3 in a sequence table.

3. The application of the DNA molecule shown by the 2310-5997 th nucleotide of the sequence 3 in the sequence table in reducing the seed shattering property and/or increasing the seed length of plants.

4. A method for cultivating transgenic plants is to inhibit the expression of protein coding genes shown in a sequence 1 in a sequence table in a target plant to obtain transgenic plants; the transgenic plant satisfies the phenotypes of (c1) and/or (c2) as follows:

(c1) the seed shattering is lower than that of the target plant;

(c2) the kernel length is larger than that of the target plant.

5. The method of claim 4, wherein: the encoding gene is a DNA molecule as described in any one of (b1) to (b3) below:

6. A method of reducing plant shattering and/or increasing kernel length comprising the steps of: reducing the expression quantity of the protein shown in the sequence 1 in the sequence table in the target plant, reducing the plant falling property and/or increasing the seed length.

7. A method for cultivating transgenic plants is to introduce coding genes of protein shown in a sequence 1 in a sequence table into target plants to obtain transgenic plants; the transgenic plant satisfies the phenotypes of (d1) and/or (d 2):

(d1) the seed shattering property is higher than that of the target plant;

(d2) the kernel length is less than that of the target plant.

8. The method of claim 7, wherein: the encoding gene is (b1) or (b2) as follows:

(b2) a DNA molecule shown as 2310-5997 th nucleotide of a sequence 2 in a sequence table.

9. A method of increasing plant shatter and/or decreasing kernel length comprising the steps of: increasing the expression quantity of the protein shown in the sequence 1 in the sequence table in the target plant, increasing the plant falling property and/or reducing the seed length.

10. Use of the method of any one of claims 4 to 9 in plant breeding.