CN107418958B - Rice RCN20 gene and its coding protein and application - Google Patents

Abstract

The invention relates to a rice RCN20 gene and its coded protein, the CDS sequence of the gene is shown in SEQ ID No.2, and the protein sequence is shown in SEQ ID No. 3. The gene is expressed in multiple tissues of rice, and the expression level is highest in tillering buds and leaf sheaths. Experiments show that the RCN20 gene mutant enables rice to show that the tillering number is reduced seriously, only 3 to 4 tillering are produced, and lateral meristem of the mutant can be formed normally in a seedling stage; in the mature period, tillering buds are formed but cannot be elongated, the cell shapes of the tillering buds are irregular, and the cell shapes of wild-type tillering buds are regular. Transformation of the rice rcn20 mutant revealed that the mutant oligotillering phenotype could be restored. The RCN20 gene can directly control the tillering number of rice by regulating the shape of cells, so that the RCN20 gene is expected to be applied to the directional design of rice plant shapes, can be used as an important gene resource and is beneficial to the improvement of rice yield.

Description

Rice RCN20 gene and its coding protein and application

Technical Field

The invention belongs to the field of crop molecular biology, and particularly relates to a rice RCN20 gene, a protein coded by the gene and application of the gene in regulation and control of rice tillering number.

Background

Rice is one of the most important food crops, and the rice is taken as the staple food by half of people in the world. In the 21 st century, agricultural land use is sharply reduced, population is continuously increased, and food shortage is serious. To solve this crisis, it is important to increase rice yield. With the completion of the rice genome-wide program, many genes and QTLs (quantitative trait loci) associated with various important agronomic traits were mapped. The research progresses lay a foundation for improving the yield of the rice and provide precious breeding materials and genetic resources.

Tillering is one of important agronomic traits determining the yield of rice, is a special branch trait formed in the growth and development process of monocotyledons, and has a certain rule. The early and late tillering of rice is related to the characteristics of the variety, and the tillering growth speed also has difference among the varieties, namely genetic factors are one of the main reasons for influencing the dynamic change of tillering. From the breeding perspective, the rice variety with good yield is characterized by strong tillering capacity, high ear forming rate, large number of ears and high maturing rate, which is also the guarantee of realizing high yield and high quality of super rice, namely, the harvest index is improved on the basis of higher biomass of super rice.

The formation process of rice tillering can be divided into two main steps, i.e. the formation of tillering buds and the elongation of tillering buds. Usually, a tillering bud is formed in each rice leaf axilla, but only the tillering bud positioned on the non-elongated internode at the base part of the stem can be elongated to form tillering; and the tillering bud at the upper part of the stem, which extends between the joints, is not extended generally and is in a dormant state all the time. Thus, the number of tillers produced by rice depends not only on the number of tillering buds that can be formed, but also on the number of tillering buds that can be elongated. The breeding of the variety with stable tillering change, long effective tillering period and high earning rate can generate enough population quantity, ensure good population structure and better coordinate the contradiction between the population and the individual, thereby further improving the rice yield.

^{TAD1 TE}The first discovered key gene regulating rice Tillering is MOC1, a homologous gene of tomato And arabidopsis thaliana derived (LS/LAS) suppressor gene, MOC1 belongs to the GRAS family, encodes a transcriptional regulator, is expressed in Tillering shoot, regulates initiation And growth of Tillering shoot at different stages of vegetative growth And reproductive growth of axillary meristem, thus, rice mutant MOC1 almost completely loses Tillering ability, only 1 main stem, no Tillering appears, spikelet on panicle is also significantly reduced, demonstrates MOC1 plays a positive role in rice Tillering And ear branching, recently, two new genes, TAD1(Tillering And dferf 1) And (tiller enhacer), are discovered at leaf axis, are co-expressed with MOC1, regulate rice Tillering And branching, TAD1 is a mutant of rice Tillering And branching, a rice transgenic line expressing a rice plant promoter, a rice transgenic line expressing a rice plant protein, a rice transgenic line expressing a protein expressing a rice transgenic line expressing a rice transgenic line expressing a rice.

The existing research results show that the strigolactone is used as a phytohormone to inhibit plant branching. It is synthesized from carotenoid precursors through the action of a series of enzymes, and four Arabidopsis genes have been mapped to participate in the control of strigolactones. Among them, MAX1 encodes a cytochrome P450 enzyme, MAX2 is a member of the F-box LRR family, MAX3 and MAX4 encode the Carotenoid Cleaving Dioxygenase (CCD) family proteins CCD7 and CCD8, respectively. MAX1, MAX3 and MAX4 are all involved in the biosynthesis of strigolactones, and MAX2 senses strigolactone signals. The rice homologs MAX2, MAX3, MAX4 have been defined by FENBIE as D3, D17/HTD and D10, respectively. Wherein D17 encodes carotenoid-cleaving dioxygenase 7(carotenoid cleavage dioxygenase 7, CCD7), D10 encodes carotenoid-cleaving dioxygenase 8(carotenoid cleavage dioxygenase8, CCD8), and homologous chromosomes of MAX1 have not been found in rice so far. However, two newly located rice genes, D27 and D14, demonstrated novel components of strigolactone biosynthesis and signaling pathways. D27 is expressed in vascular bundle tissue and encodes an alpha/beta-carotenoid isomerase. D14 encodes a hydrolase/esterase 4, which may be involved in strigolactone perception.

A great breakthrough in the aspect of illustrating rice tillering is to find the strigolactone which is terpenoid lactone capable of inhibiting the growth of tillering buds. From this discovery, new mutants D27 and D14 have been identified by reduced strigolactone levels and responses. D27 and D14 may each play important roles in strigolactone biosynthesis and signal transduction. There are many axillary growth and dwarfing mutants in rice and Arabidopsis thaliana. These mutants have similar phenotypes, increased branching and shortened plant strains, and the comparative experiments between rice and Arabidopsis have found that the phenotypes of the mutants are due to deletion of strigolactone synthesis and signals. Strigolactone, an terpenoid lactone, is the end product of the MAX/RMS/D pathway, inhibiting the growth of tillering buds.

The rice TB1 gene (OsTB1) was cloned from the similar sequence of the maize BRANCHED regulatory gene TEOSINTE BRANCHED 1(TB 1). Both genes encode putative transcription factors that have a basic helix-loop-helix DNA binding motif, designated TCP domains. The genetic locus of OsTB1 demonstrated to be a homologous gene to maize TB 1. We found that overexpression of OsTB1 reduced tillering number, while tillering increased in FC1 mutant and OsTB1 function was absent. These results show that OsTB1 is involved in the negative regulation of rice lateral branching, similar to the TB1 gene in maize. The lateral branch consists of two parts, tillering bud formation and tillering bud elongation. OsTB1 may play an important role in regulating and controlling the elongation of tillering buds, and tillering primordium of a transgenic plant overexpressed in OsTB1 can still normally reproduce. OsTB1 was expressed throughout the tillering shoot as shown by the promoter-GUS fusion gene, but could inhibit only the late elongation.

The rice RCN20 gene belongs to a beta-tubulin family, the beta-tubulin family in rice has 8 genes which are divided into 4 types, TUB1, TUB4 and TUB6 are the first type; TUB3, TUB5, TUB7 are of the second class; TUB2 is of the third type; TUB8 is of the fourth type. RCN20 is TUB 2. Alpha-tubulin and beta-tubulin are the major components that make up microtubules, and their basic units, tubulin heterodimers, that make up microtubules, form microtubules, and these two types of proteins need to bind GTP first and then to the (+) terminal (extension) of microtubules; after becoming part of microtubules, GTP bound to tubulin is hydrolyzed to GDP. Although both of these proteins can bind GTP, only β -tubulin has gtpase activity and can hydrolyze bound GTP to GDP, whereas α -tubulin does not. Whether β -tubulin binds GTP or GDP in the tubulin dimer affects the stability of the dimer in microtubules. GTP-bound dimers tend to form microtubules, while GDP-bound dimers tend to dissociate from microtubules; thus, such a cycle of GTP and GDP constitutes the dynamic equilibrium of microtubules. At present, no research report on the RCN20 gene of rice and the function thereof exists.

Disclosure of Invention

The invention aims to provide a rice RCN20 gene and a coding protein and application thereof.

The rice RCN20 gene provided by the invention has the CDS sequence:

1) A nucleotide sequence shown as SEQ ID No. 2; or

2) The nucleotide sequence shown in SEQ ID No.2 is substituted, deleted and/or added with one or more nucleotides; or

3) A DNA sequence which hybridizes with the nucleotide sequence defined in 1) under stringent conditions.

The invention provides a protein coded by rice RCN20 gene, which comprises the following components:

1) An amino acid sequence shown as SEQ ID No. 3; or

2) Protein which is derived from the protein 1) and has the same functional activity and is obtained by substituting, deleting and/or adding one or more amino acids in the amino acid sequence shown in SEQ ID No. 3.

The invention provides a biological material containing the rice RCN20 gene, which is a carrier, a host cell and a transformed plant cell.

the invention provides application of the rice RCN20 gene or the protein coded by the gene in preparing transgenic plants.

The plant is rice.

The invention provides application of the rice RCN20 gene or the protein coded by the gene in rice germplasm resource improvement.

The invention provides application of the rice RCN20 gene or the protein coded by the gene in improving rice yield.

The invention provides the application of the rice RCN20 gene or the protein coded by the gene in crop improvement breeding and seed production.

The crops are rice.

The invention provides application of the rice RCN20 gene or the protein coded by the gene in promoting the shape regulation of tillering bud cells of crops.

The invention provides application of the rice RCN20 gene or the protein coded by the gene in increasing the tillering number of crops.

The invention has the advantages that: the invention discovers that the rice RCN20 gene is expressed in a plurality of tissues of rice, wherein the expression level is the highest in tillering buds and leaf sheaths. Experiments show that the rice RCN20 gene can directly influence the tillering number of rice after mutation. RCN20 is transformed into rice RCN20 mutant, and the phenotype of the normal tillering number of the mutant can be recovered, so that the RCN20 gene can directly regulate and control the tillering number and be applied to the control of the plant shape of the rice, and the cell shape of the tillering bud can be promoted to be regular, so that the rice productivity is improved.

Drawings

FIG. 1 shows the phenotype of rcn20 mutant versus wild-type Nissan rice 808. Fig. 1A, 1B, 1C show that the mutants only have 3 to 4 tillers. The rice rcn20 mutant tillering bud could form (fig. 1D, arrows indicate tillering buds, where the left panel is the wild type at six-leaf one-heart stage, the right panel is mutant rcn20), but some could not elongate at the later stage (fig. 1E, upper layer is wild type, lower layer is mutant rcn 20; 1F, left is low-node tillering of wild type, right is low-node tillering of mutant rcn 20; 1G, left is high-node tillering of wild type, right is high-node tillering of mutant rcn20, where arrows indicate both tillering buds).

FIG. 2 is a diagram showing the location and structure of RCN20 gene.

FIGS. 3A and 3B show the expression pattern analysis and GUS staining of RCN20 gene in rice tissues, respectively.

FIG. 4 is the structural diagram of vector pCAMBIA1305.1: RCN 20.

FIG. 5 is the structural diagram of vector pCAMBIA1305.1-APFHN: RCN 20.

FIG. 6 shows pCAMBIA1305.1-APFHN:RCN 20 transformed rice RCN20 mutant can restore its phenotype. FIG. 6A shows that RCN20 genome DNA is transferred into rice RCN20 mutant, the number of the mutant tillers is recovered to be normal, and the number of the mutant tillers is as much as that of the wild type, wherein Line1 and Line2 respectively refer to transgenic lines with normal tillering numbers; FIG. 6B shows that the rice mutant RCN20 has significantly increased tillering number and is slightly more than the wild type, where OE-1 and OE-2 refer to transgenic lines with over-expressed genes, respectively, after the over-expressed RCN20 genomic DNA is transferred into the rice mutant RCN 20.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention.

Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art; all reagents used in the examples are commercially available unless otherwise specified.

The rice (Oryza sativa) variety Sheng Rice 808 used in the examples was a standard variety, and the rice few-tillering mutant rcn20 was from the institute of crop science, Chinese academy of agricultural sciences.

Example 1 acquisition and phenotypic analysis of Rice rcn20 mutants

The japonica rice variety Sheng rice 808 is subjected to chemical mutagenesis by EMS to obtain an oligotiller variant rcn20(reduced culm number-20). In the rice rcn20 mutant, the 2 nd exon of the rcn20 gene, namely the 616 th nucleotide of the coding region of the gene, is mutated from G to A, so that the 206 th amino acid is mutated from alanine to threonine.

Phenotypic analysis shows that the height of the rice rcn20 mutant plant is shortened by 32.2% compared with the wild type, and the tillering number of the rcn20 mutant is obviously reduced and is only 3-4 tillering (fig. 1A, 1B and 1C). The rice rcn20 mutant tillering bud could form (fig. 1D, where a is the wild type at six-leaf one-heart stage, b is mutant rcn20), but some could not elongate at later stage (fig. 1E, upper layer is wild type, lower layer is mutant rcn 20; fig. 1F, left is the low-node tillering of wild type, right is the low-node tillering of mutant rcn 20; fig. 1G, left is the high-node tillering of wild type, right is the high-node tillering of mutant rcn 20). The lateral meristem of the mutant can be normally formed in the seedling stage; in the mature period, tillering buds are formed but cannot be elongated, the cell shapes of the tillering buds are irregular, and the cell shapes of wild-type tillering buds are regular. The mutant has only 3 to 4 tillers, the plant is slightly short, the root is umbrella-shaped, and the root length is seriously shortened.

Example 2 acquisition of Rice rcn20 Gene and functional complementation test

The RCn20 mutant and the indica rice variety Dular with normal phenotype and high polymorphism are hybridized to obtain an F ₂ segregation population, genetic analysis and gene location are carried out, and the strain analysis of character segregation of the F2 generation shows that both the normal individual plant and the mutant individual plant accord with the segregation ratio of 3:1, so that the mutant character is controlled by a pair of recessive genes.

30 mutants of F2 are used as materials, 170 Indel markers uniformly distributed on 12 rice chromosomes are used for positioning candidate genes on the No.3 chromosome, the candidate genes are linked with Indel markers M2 and M3, the physical distance between the two markers is 300Kb (an A diagram of a figure 2), in order to further finely position the candidate genes, the F ₂ generation positioning population is expanded to 202 strains, new markers M6, M7 and M8 are developed for linkage analysis, recombination events of target genes at the markers are found to be 10, 2 and 6 respectively, the candidate genes are positioned between M7 and M8, and the physical distance between the two markers is about 87 Kb.

There were 8 genes between markers M7 and M8 (FIG. 2, panel B), according to gene annotation information provided by the TIGR website (http:// rice. plant. msu. edu /). Wherein the gene sequence (FIG. 2, panel C) of LOC _ Os03g01530 has a gene function related to phenotype, and the full-length genomic DNA of the gene is subjected to PCR amplification. The primers used are shown in Table 1, and the sequencing results of the wild type and the mutant were analyzed using DNAStar software. In the rcn20 mutant, the mutation from G to A in exon 2 of the gene, i.e., the 616 th nucleotide of the coding region of the gene, resulted in the mutation from alanine to threonine in amino acid 206 (FIG. 2, panel D). RCN20 genome DNA total length of about 1811bp (SEQ ID NO.1), CDS total length 1344bp (shown as SEQ ID NO.2 sequence), containing 2 exons, 1 intron, encoding a protein product (shown as SEQ ID NO.3 sequence) consisting of 447 amino acids.

TABLE 1 primer sequences referred to in example 2

Example 3 Rice RCN20 Gene expression Pattern

In order to determine the tissue expression mode of the RCN20 gene, the expression level of the gene in each tissue of rice including roots, stems, leaves, leaf sheaths, scions and tillering buds is detected by a Real-time PCR method, and the result shows that the RCN20 gene is expressed in the tissues of roots, stems, leaves, leaf sheaths, scions and tillering buds of rice, wherein the expression level is highest in the tillering buds, higher in the leaf sheaths and relatively lower in the leaves and the stems (see fig. 3A and 3B).

TABLE 2 primer sequences related to this example

Example 4 pcambial 305.1: : transformation of rice RCN20 mutant with RCN20 vector

In order to carry out a function complementation experiment, an RCN20 gene function complementation vector driven by a self promoter and an over-expression vector driven by a rice ACTIN1 promoter are respectively constructed.

The RCN20 gene function complementary vector is driven by gene self promoter, 2169bp before ATG translation initiation site is selected as gene promoter, 604bp after TAA, including 251bp of complete 3' -UTR. EcoRI site is introduced into the 5 'end, PmlI site is introduced into the 3' end, and the size of PCR product is 4193 bp. Recombination into EcoRI + PmlI site of pCAMBIA1305.1 constitutes a reversion vector driven by self-promoter (FIG. 4). In addition, a plant binary expression vector pCAMBIAl305.1-APFHN was used to construct an overexpression vector, and PCR amplification was performed using cDNA as a template, using amplification primers 03g01530CDS as shown in Table 3. The SalI site was introduced at both the 5 ' and 3 ' ends, and contained 54bp 3 ' -UTR, and the PCR product length of 1398bp. was recombined into the SalI site of pCAMBIA1305.1-APFHN. Driven by a constitutive high-expression rice Actin1 promoter. The constructed vector is shown in FIG. 5.

The constructed complementary vector and the overexpression vector are transferred into agrobacterium EHA105 by an electric shock method, and the seed induction callus of the rice rcn20 mutant knot is used as a receptor material to carry out the transformation of rice by an agrobacterium-mediated transformation method. The functional complementation vector driven by its own promoter yielded 5 independent transformed lines, 4 of which were restored to wild type phenotype. Whereas the overexpression vector driven by the ACTIN1 promoter yielded 7 independent transformation lines, 6 of which were restored to wild-type phenotype (FIG. 6A). The tillering of the over-expressed transgenic plants is counted, and the result shows that the tillering number of the transgenic plants is obviously more than that of the mutant and slightly more than that of the wild type, and the height of the plants is also consistent with that of the wild type (figure 6A). These results indicate that it is indeed due to the mutation of the RCN20 gene that reduced the number of tillers of the RCN20 mutant.

TABLE 3 primer sequences related to this example

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the technical principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Sequence listing

<110> institute of crop science of Chinese academy of agricultural sciences

<120> rice RCN20 gene and its coding protein and application

<160> 27

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1811

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 1

aacactccac taacgcctcc cgacgccgcc gccgcctcct cctcttgttt cgccaccgcc 60

gccgccgcct acgaggaaga agaaagaaga ggaaggagaa gatgagggag atcctccaca 120

tccagggtgg gcaatgcggg aaccagatcg gcgccaagtt ctgggaggtg gtgtgcgcgg 180

agcacgggat cgacgccacc gggcgctacg acggcgactc cgacctgcag ctggagcgcg 240

tcaacgtgta ctacaatgag gcctcctgcg ggcgcttcgt cccgcgcgcc gtgctcatgg 300

acctggagcc gggcaccatg gactccgtcc gctccggccc ctacggccac atcttccgcc 360

cggacaactt cgtcttcggc cagtccggcg ccggcaacaa ctgggccaag ggacactaca 420

ccgagggcgc cgagctcatc gacgccgtcc tcgacgtcgt acgaaaggag gccgagaact 480

gcgactgcct ccaaggtaac tagtacccta actatctcag atctgcctct cctcaaatca 540

aatctgagtt caaatgctgc tgctgctgca gggttccagg tttgccactc gcttggcggc 600

ggcacggggt ccggcatggg cacgctcctc atctccaaga tccgggagga gtaccccgac 660

cggatgatgc tcaccttctc cgtcttcccc tcgcccaagg tctccgacac cgtggtggag 720

ccctacaacg ccaccctctc cgtccaccag ctcgtcgaga acgccgacga gtgcatggtg 780

ctcgacaacg aggccctcta cgacatctgc ttccgcaccc tcaagctcac caccccaagc 840

ttcggcgacc tcaaccacct catctccgcc accatgagcg gcgtcacctg ctgcctccgc 900

ttccccggcc agctcaactc cgacctccgc aagctcgccg tcaacctcat ccccttccct 960

cgcctccact tcttcatggt cggcttcgcc ccgctcacct cccgcggctc gcagcagtac 1020

cgcgccctca ccgtcccgga gctcacccag cagatgtggg acgccaagaa catgatgtgc 1080

gccgccgacc cccgccacgg ccgctacctc accgcctccg ccatgttccg cggcaagatg 1140

tccaccaagg aggtggacga gcagatgctc aacgtccaga acaagaactc ctcctacttc 1200

gtcgagtgga tccccaacaa tgtcaagtcc accgtctgcg acatcccccc caccggcctc 1260

aagatggcct ccaccttcat cggcaactcc acctccatcc aggagatgtt ccgccgcgtc 1320

agcgagcagt tcaccgccat gttccgcagg aaggccttct tgcactggta cacgggcgag 1380

ggcatggacg agatggagtt caccgaggct gagagcaaca tgaacgacct cgtctccgag 1440

taccagcagt accaggacgc caccgccgac gacgagggtg aatacgagga tgaggaagaa 1500

gaggctgatc tccaggacta attactactc tactgcaata tcaatgtcca atctaatggg 1560

gttcggttca cttcttgtgt tgcttaggct atcttctcaa gtgttgtcgt tcttaattat 1620

tactcctagc tgctactcta ctctacttga tgtattatta ttactcagct ttgattgatg 1680

ataggcacaa taccctgttg ccttgtactc gtaggagtat gtgctgttac tgtctaactg 1740

atctgtatga attatcatcc tatttttgtt aatctatctt ctagtaaaat gcagagttgc 1800

tgcttagagt a 1811

<210> 2

<211> 1341

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 2

atgagggaga tcctcccatc cagggtgggc aatgcgggaa ccagatcggc gccaagttct 60

gggaggtggt gtgcgcggag cacgggatcg acgccaccgg gcgctacgac ggcgactccg 120

acctgcagct ggagcgcgtc aacgtgtact acaatgaggc ctcctgcggg cgcttcgtcc 180

cgcgcgccgt gctcatggac ctggagccgg gcaccatgga ctccgtccgc tccggcccct 240

acggccacat cttccgcccg gacaacttcg tcttcggcca gtccggcgcc ggcaacaact 300

gggccaaggg acactacacc gagggcgccg agctcatcga cgccgtcctc gacgtcgtac 360

gaaaggaggc cgagaactgc gactgcctcc aagggttcca ggtttgccac tcgcttggcg 420

gcggcacggg gtccggcatg ggcacgctcc tcatctccaa gatccgggag gagtaccccg 480

accggatgat gctcaccttc tccgtcttcc cctcgcccaa ggtctccgac accgtggtgg 540

agccctacaa cgccaccctc tccgtccacc agctcgtcga gaacgccgac gagtgcatgg 600

tgctcgacaa cgaggccctc tacgacatct gcttccgcac cctcaagctc accaccccaa 660

gcttcggcga cctcaaccac ctcatctccg ccaccatgag cggcgtcacc tgctgcctcc 720

gcttccccgg ccagctcaac tccgacctcc gcaagctcgc cgtcaacctc atccccttcc 780

ctcgcctcca cttcttcatg gtcggcttcg ccccgctcac ctcccgcggc tcgcagcagt 840

accgcgccct cacccagcag atgtgggacg ccaagaacat gatgtgcgcc gccgaccccc 900

gcaccgtccc ggagctccac ggccgctacc tcaccgcctc cgccatgttc cgcaaggagg 960

tggacgagca gatgctcaac gtccagaaca agaactcctc ctacggcaag atgtccactt 1020

cgtcgagtgg atccccaaca atgtcaagtc caccccccca ccggcctcaa gatggcctcc 1080

accttcatcg gcaactccac ctccgtctgc gacatccatc caggagatgt tccgccgcgt 1140

cagcgagcag ttcaccgcca tgttccgcag gaaggccttc ttgcactggt acacgggcga 1200

gggcatggac gagatggagt tcaccgaggc tgagagcaac atgaacgacc tcgtctccga 1260

gtaccagcag taccaggacg ccaccgccga cgacgagggt gaatacgagg ataggaagaa 1320

gaggctgatc tccaggacta a 1341

<210> 3

<211> 447

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 3

Met Arg Glu Ile Leu His Ile Gln Gly Gly Gln Cys Gly Asn Gln Ile

1 5 10 15

Gly Ala Lys Phe Trp Glu Val Val Cys Ala Glu His Gly Ile Asp Ala

20 25 30

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

35 40 45

Val Tyr Lys Gly His Tyr Thr Glu Gly Ala Glu Leu Ile Asp Ala Val

50 55 60

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

65 70 75 80

Phe Gln Val Cys His Ser Leu Gly Gly Gly Thr Gly Ser Gly Met Gly

85 90 95

Thr Leu Leu Ile Ser Lys Ile Arg Glu Glu Tyr Pro Tyr Asn Glu Ala

100 105 110

Ser Cys Gly Arg Phe Val Pro Arg Ala Val Leu Met Asp Leu Glu Pro

115 120 125

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

130 135 140

Pro Asp Asn Phe Val Phe Gly Gln Ser Gly Ala Gly Asn Asn Trp Ala

145 150 155 160

Asp Arg Met Met Leu Thr Phe Ser Val Phe Pro Ser Pro Lys Val Ser

165 170 175

Asp Thr Val Val Glu Pro Tyr Asn Ala Thr Leu Ser Val His Gln Leu

180 185 190

Val Glu Asn Ala Asp Glu Cys Met Val Leu Asp Asn Glu Ala Leu Tyr

195 200 205

Asp Ile Cys Phe Arg Thr Leu Lys Leu Thr Thr Pro Ser Phe Gly Asp

210 215 220

Leu Asn His Leu Ile Ser Ala Thr Met Ser Gly Val Thr Cys Cys Leu

225 230 235 240

Arg Phe Pro Gly Gln Leu Asn Ser Asp Leu Arg Lys Leu Ala Val Asn

245 250 255

Leu Ile Pro Phe Pro Arg Leu His Phe Phe Met Val Gly Phe Ala Pro

260 265 270

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

275 280 285

Leu Thr Gln Gln Met Trp Asp Ala Lys Asn Met Met Cys Ala Ala Asp

290 295 300

Pro Arg His Gly Arg Tyr Leu Thr Ala Ser Ala Met Phe Arg Gly Lys

305 310 315 320

Met Ser Thr Lys Glu Val Asp Glu Gln Met Leu Asn Val Gln Asn Lys

325 330 335

Asn Ser Ser Tyr Phe Val Glu Trp Ile Pro Asn Asn Val Lys Ser Thr

340 345 350

Val Cys Asp Ile Pro Pro Thr Gly Leu Lys Met Ala Ser Thr Phe Ile

355 360 365

Gly Asn Ser Thr Ser Ile Gln Glu Met Phe Arg Arg Val Ser Glu Gln

370 375 380

Phe Thr Ala Met Phe Arg Arg Lys Ala Phe Leu His Trp Tyr Thr Gly

385 390 395 400

Glu Gly Met Asp Glu Met Glu Phe Thr Glu Ala Glu Ser Asn Met Asn

405 410 415

Asp Leu Val Ser Glu Tyr Gln Gln Tyr Gln Asp Ala Thr Ala Asp Asp

420 425 430

Glu Gly Glu Tyr Glu Asp Glu Glu Glu Glu Ala Asp Leu Gln Asp

435 440 445

<210> 4

<211> 18

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 4

gcccaagcca acgtcgtc 18

<210> 5

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 5

agaggaacaa tcaaccagac aa 22

<210> 6

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 6

gcatgttgta ctcctcctta ct 22

<210> 7

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 7

atgcatgggt tctcaagtga 20

<210> 8

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

cagatcctat ggcccctgag 20

<210> 9

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 9

caggtgtttg acggttatgg t 21

<210> 10

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 10

gaagagagta ccggaagcga 20

<210> 11

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 11

cttgtgtgtg ttttgcgctc 20

<210> 12

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 12

actgcaagga ttaaggggct 20

<210> 13

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 13

aaggggctgt tcggattgta 20

<210> 14

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 14

tcagctgaac ctcctgtagc 20

<210> 15

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 15

aaaggtggtt ggctagacga 20

<210> 16

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 16

atgggacatg cagctcactc a 21

<210> 17

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 17

tcgttccgta tttttacccg a 21

<210> 18

<211> 19

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 18

ttcgaattcc ctctgctcg 19

<210> 19

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 19

cagtggtcag tggcagagaa 20

<210> 20

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 20

caccctggct gactacaaca 20

<210> 21

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 21

ttcttcttgc ggcagttgac 20

<210> 22

<211> 18

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 22

atgggcacgc tcctcatc 18

<210> 23

<211> 18

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 23

gaagcagatg tcgtagag 18

<210> 24

<211> 39

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 24

ccatgattac gaattctgca accacatttg acattttct 39

<210> 25

<211> 40

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 25

gtcaccaatt cacacgtgtg atcaagtggc tgtatgatcg 40

<210> 26

<211> 37

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 26

ccatgattac gaattcatga gggagatcct ccacatc 37

<210> 27

<211> 41

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 27

cccttgctca ccatggtacc gtcctggaga tcagcctctt c 41

Claims (2)

1. The rice RCN20 gene mutant is characterized in that , of the No.2 exon of a CDS sequence of a gene sequence with the TIGR website number LOC _ Os03G01530, namely the No. 616 nucleotide of a coding region of the gene, is mutated from G to A, so that the No. 206 amino acid of the corresponding coding amino acid sequence is mutated from alanine to threonine.

2. A biomaterial containing the rice RCN20 gene mutant according to claim 1, wherein the biomaterial is a vector, a host cell incapable of being propagated into an animal plant species, or a transformed plant cell incapable of being propagated into a plant species.