CN108586593A - With the relevant albumen of rice seed holding and its encoding gene and application - Google Patents

Abstract

The invention discloses a kind of applications with the relevant albumen of rice seed holding and its encoding gene.Albumen provided by the present invention is following any：A) protein that amino acid sequence forms shown in SEQ ID No.1；B) amino acid sequence defined by a) by the substitution of one or several amino acid residues and/or is lacked and ored add, and with the relevant protein of the seed holding of vegetable seeds；C) and a) b) in it is any defined by amino acid sequence there is 95% or more, 90% or more, 85% or more or 80% or more homology, and with the relevant protein of the seed holding of vegetable seeds.SH3 albumen and its encoding gene provided by the present invention are of great significance in terms of the seed holding of regulation and control vegetable seeds, will play a significant role in cultivating high yield new variety of plant.

Description

With the relevant albumen of rice seed holding and its encoding gene and application

Technical field

The invention belongs to plant genetic engineering fields, are related to a kind of and the relevant albumen of rice seed holding and its encoding gene Application.

Background technology

Oryza (Oryza L.) is containing there are two cultigens, i.e. Asian Cultivated Rice (O.sativa L.) and Oryza glaberrima Steud (Oryza glaberrima Steud.), they independently originate from Asia and Africa.Asian Cultivated Rice yield is high, alive Boundary is planted various regions extensively, and Oryza glaberrima Steud yield is relatively low, is only limitted to West Africa area.

Oryza glaberrima Steud be before about 3000 independently originate from Africa cultivated rice and Asian Cultivated Rice be by different Progenitor species are tamed.Asian Cultivated Rice is just introduced West Africa area, is planted so as to cause Africa by 16 middle of century, Portuguese The species of training rice are drastically reduced, so the cultivated area of Oryza glaberrima Steud is much smaller than Asian Cultivated Rice now.Currently, Asia is cultivated Rice plantation extensively in the world, food source is provided for global nearly half population.Oryza glaberrima Steud is distributed mainly on non- Continent is western, is one of local main cereal crops, and positive effect is played to the civilization and economy in Africa.However, with Asia Continent cultivated rice is compared, and the correlative study of Oryza glaberrima Steud is very deficient, is gone through to the population genetic variations and evolution of Oryza glaberrima Steud History is known little about it.

Since Oryza glaberrima Steud has the characteristics that growth is fast, impoverishment tolerant, drought-resistant, Productive statistics are low, so special There is certain production advantage in ecological environment.Therefore, further deepen the research to Oryza glaberrima Steud to plant for improving Africa The yield of training rice plays an important roll.

Seed holding is the rice varieties of the important character one of closely related with Rice Production, too easy shattering or too difficult shattering It should not all be applied in production.Excavation with rice seed holding related gene will be to understand rice domestication to accumulate new material, New genetic resources are also provided for Breeding Application.

Invention content

The object of the present invention is to provide a kind of applications with the relevant albumen of rice seed holding and its encoding gene.

In a first aspect, a kind of claimed protein.

Protein provided by the present invention is named as SH3, concretely following any：

A) protein that amino acid sequence forms shown in SEQ ID No.1；

B) by amino acid sequence defined by a) by the substitution of one or several amino acid residues and/or missing and/or Addition, and with the relevant protein of the seed holding of vegetable seeds；

C) and a)-b) in it is any defined by amino acid sequence have 95% or more, 90% or more, 85% or more or 80% or more homology, and with the relevant protein of the seed holding of vegetable seeds.

Wherein, b) and c) shown in protein can be from rice protein.

For the ease of the purifying of the protein, can be connected in the amino terminal or carboxyl terminal of the protein upper as follows Label shown in table.

Table：The sequence of label

Label	Residue	Sequence
			Poly-Arg	5-6 (being usually 5)	RRRRR
Poly-His	2-10 (being usually 6)	HHHHHH
			FLAG	8	DYKDDDDK
Strep-tag II	8	WSHPQFEK
			c-myc	10	EQKLISEEDL

Second aspect, the nucleic acid molecules of claimed code for said proteins.

The nucleic acid molecules can be DNA, such as cDNA, genomic DNA or recombinant DNA；The nucleic acid molecules can also be RNA, such as mRNA, hnRNA or tRNA.

In one embodiment of the invention, the nucleic acid molecules are specially that the gene of code for said proteins (is named as SH3 genes), the gene concretely it is following it is any shown in DNA molecular：

1) DNA molecular shown in 2209-11125 of SEQ ID No.2；

2) DNA molecular shown in SEQ ID No.3；

3) under strict conditions with 1) -2) in the DNA molecular of any restriction hybridize and encode protein described in claim 1 DNA molecular；

4) with 1) -3) in any restriction DNA sequence dna have 95% or more, 90% or more, 85% or more or 80% with Upper homology, and the DNA molecular of code for said proteins.

Above-mentioned stringent condition can be with 6 × SSC, and the solution of 0.5%SDS hybridizes at 65 DEG C, then with 2 × SSC, It is primary that 0.1%SDS and 1 × SSC, 0.1%SDS respectively wash film.

Wherein, 2209-11125 of SEQ ID No.2 are sequence of the SH3 genes in rice genome；SEQ ID No.3 is its cDNA sequence (CDS).SEQ ID No.2 and SEQ ID No.3 encode protein shown in SEQ ID No.1.

The third aspect, the claimed recombinant vector containing the nucleic acid molecules, expression cassette, transgenic cell line Or recombinant bacterium.

Wherein, the recombinant vector can be recombinant expression carrier or recombinant cloning vector.

The recombinant expression carrier can use existing plant expression vector construction.The plant expression vector includes double base agriculture Bacillus carrier and the carrier etc. that can be used for plant micropellet bombardment, as pCAMBIA1300, pCUbi1390, pCHF3, PGreen0029, pCAMBIA3301, pBI121, pBin19, pCAMBIA2301, pCAMBIA1301-UbiN or other are derivative to plant Object expression vector.The plant expression vector also may include 3 ' end untranslated regions of foreign gene, that is, include that polyadenylic acid is believed Number and it is any other participation mRNA processing or gene expression DNA fragmentation.The bootable polyadenylic acid of polyadenylation signals adds Enter the 3 ' ends to mRNA precursor.When using the gene constructed recombinant expression carrier, it can be added before its transcription initiation nucleotide Any type is enhanced, composing type, organizing specific type or inducible promoter, such as cauliflower mosaic virus (CAMV) 35S is opened Mover, ubiquitin gene Ubiquitin promoters (pUbi), stress induced promoter rd29A etc., they can be used alone or with Other plant promoters are used in combination；In addition, when using gene constructed recombinant expression carrier of the invention, enhancing also can be used Son, including translational enhancer or transcriptional enhancer, these enhancer regions can be ATG initiation codon or neighboring region starting Codon etc., but must be identical as the reading frame of coded sequence, to ensure the correct translation of entire sequence.The translation control letter Number and the source of initiation codon be extensive, can be natural, can also be synthesis.Translation initiation region can come from Transcription initiation region or structural gene.It, can be to used for the ease of transgenic plant cells or plant are identified and screened Recombinant expression carrier is processed, and the enzyme or luminophor of color change can be generated as the coding that can be expressed in plant is added Gene, resistant antibiotic marker or anti-chemical reagent marker gene etc..Also any selected marker can be not added with Gene directly screens transformed plant with adverse circumstance.

In the present invention, the promoter for starting the genetic transcription in the recombinant expression carrier is the endogenous startup of rice Son.

The expression cassette is by that can start the promoter of the gene expression, the gene and transcription terminator group At.

Fourth aspect, claimed protein described previously or the nucleic acid molecules or the recombinant vector, Application in the seed holding of expression cassette, transgenic cell line or recombinant bacterium regulation and control vegetable seeds.

In the application, the protein or the nucleic acid molecules expression quantity in the plant and/or activity are got over The seed holding of height, the plant is stronger；The protein or the nucleic acid molecules expression quantity in the plant and/or activity Lower, the seed holding of the plant is weaker.

In terms of 5th, claimed following method (A) or (B)：

(A) a kind of method reducing plant fallen, includes the following steps：Reduce the table of protein described in recipient plant Up to amount and/or activity.

(B) a kind of method improving plant fallen, includes the following steps：Improve the table of protein described in recipient plant Up to amount and/or activity.

In terms of 6th, claimed following method (C) or (D)：

(C) a kind of method for cultivating the genetically modified plants that seed holding reduces includes the steps that following (a1) and (a2)：

(a1) encoding gene of protein described in recipient plant is knocked out or inhibits to express, obtain transgenosis plant Object；

(a2) it is obtained compared with the recipient plant from genetically modified plants obtained by step (a1), what seed holding reduced turns base Because of plant.

(D) a kind of method for cultivating the genetically modified plants that seed holding improves includes the steps that following (b1) and (b2)：

(b1) encoding gene that the protein is imported into recipient plant obtains the transgenosis for expressing the encoding gene Plant；

(b2) it is obtained compared with the recipient plant from genetically modified plants obtained by step (b1), what seed holding improved turns base Because of plant.

Further, in method (C), it is described " to the encoding gene of protein described in recipient plant carry out knock out or Inhibit expression " it can be realized by any technological means that can realize this purpose, such as (such as by sequence specific nuclease CRISPR/Cas9 nucleases) specific cleavage is carried out to the encoding gene, to reduce its table in the recipient plant It reaches.

In the present invention, described " carrying out inhibition expression to the encoding gene of protein described in recipient plant " is specifically logical Cross the realization of CRISPER/Cas9 technologies；To meet 5 '-N in DNA fragmentation shown in SEQ ID No.3_X- NGG-3 ' or 5 '-CCN- N_XThe segment of -3 ' series arrangements rule is target sequence；N indicates that any one of A, G, C and T, 14≤X≤30, and X are integer, N_X Indicate X continuous deoxyribonucleotides.More specifically, in one particular embodiment of the present invention, the X is 19. Correspondingly, the target sequence is specially following any：6040-6058 of SEQ ID No.2, the of SEQ ID No.2 6056-6074,6068-6086 of SEQ ID No.2,10435-10453 of SEQ ID No.2.

Further, in method (D), " encoding gene that the protein is imported into recipient plant " can pass through The recombinant expression carrier containing the encoding gene is imported into the recipient plant to realize.

In the above-mentioned methods, it by the recombinant expression carrier for the encoding gene for carrying the protein or is used for The gene editing tool that is used when " carrying out inhibition expression to the encoding gene of protein described in recipient plant " import it is described by Body plant, concretely：By using Ti-plasmids, Ri plasmids, plant viral vector, directly delivered DNA, microinjection, conductance, Agriculture bacillus mediated equal conventional biology methods conversion plant cell or tissue, and the plant tissue of conversion is cultivated into plant.

In the present invention, all plant can be monocotyledon or dicotyledon above.Wherein, the list Cotyledon plant such as grass, it is specific such as rice, more specific such as African rice.

In one embodiment of the invention, the recipient plant is specially the wild rice varieties W1411 in Africa.

It is demonstrated experimentally that the encoding gene of protein shown in SEQ ID No.1 can be expressed in rice varieties W1411 wild to Africa Carry out CRISPR-Cas9 gene editings, compared with the plant without transgenosis under the same terms, the seed holding of transgenic paddy rice It reduces, becomes not shattering.SH3 albumen and its encoding gene provided by the present invention have in terms of the seed holding of regulation and control vegetable seeds It is significant, it will play a significant role in cultivating high-yield rice new varieties.

Description of the drawings

Fig. 1 is Africa wild rice W1411 compared with the seed holding of African rice varieties IRGC104165.

Fig. 2 is that SH3 genes (ObSH3) position schematic diagram.

Fig. 3 is W1411 in embodiment 2 and is transferred to pBWA (V) H-cas9i2 zero loads and CRISPR-Cas9 gene editings load The sequencing result of the transgenic line of body (Cas9-1, Cas9-2, Cas9-3 are respectively 3 independent transgenic lines).

Fig. 4 is W144 in embodiment 2 and is transferred to pBWA (V) H-cas9i2 zero loads and CRISPR-Cas9 gene editings load The seed holding of the transgenic line of body compares.

Specific implementation mode

Experimental method used in following embodiments is conventional method unless otherwise specified.

The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.

African rice varieties IRGC104165 and African wild rice W1411：It can be obtained from International Rice, network address：http:// irri.org/。

PBWA (V) H-cas9i2 carriers can obtain (http from Wuhan Biorun Bio-Tech. Co., Ltd.:// Www.biorun.net/), article No. corresponding to carrier is BR-cas9-i2.Be recorded in " Yongxiang Liao, Que Bai, Peizhou Xu,et al.Mutation in Rice Abscisic Acid2 Results in Cell Death, Enhanced Disease-Resistance,Altered Seed Dormancy and Development.Front.Plant Sci., 28 March, 2018. " one texts.

The positioning of embodiment 1, the African rice seed holding gene of regulation and control

Oryza glaberrima Steud kind IRGC104165 hybridizes with Africa wild rice W1411 (Fig. 1) constructs one comprising 168 The F of single plant₂Segregating population.In this F₂In group, seed holding all with molecular labeling RM5626 and MR81 close linkages, finally will The SH3 assignments of genes gene mapping are between the two labels.Then using 2650 recessive single plants in the segregating population after expanding, and according to Mark the primers between RM5626 and MR81, finally by the SH3 assignments of genes gene mapping to molecular marker SNP 29 and SNP31 this Between two labels (Fig. 2).In Oryza glaberrima Steud IRGC104165, the physical distance between the two labels is 17kb, Website http://www.softberry.com/ is upper according to Oryza glaberrima Steud IRGC96717 (CG14) sequence prediction, which does not have There is candidate gene.In African wild rice W1411, the physical distance between the two labels is 63kb, with IRGC104165 phases Than there are one 45kb segments to be inserted into this section, in website http://www.softberry.com/ is upper according to African wild rice W1411 sequence predictions have 6 open reading frame ORF in the section of this 63kb, by RT-PCR analysis shows, at this 6 Only have ORF3 to be expressed at the absciss layer position of control seed shattering in ORF, therefore the candidate gene that ORF3 is SH3, control Africa The shattering character of cultivated rice.

The acquisition and its functional verification of embodiment 2, SH3 transgenosis Africa rice

One, the acquisition of SH3 gene orders

It is as follows according to the SH3 genes of African wild rice W1411 and promoter sequence design primer, primer sequence：

F：5 '-tgcagagacttccgggttga-3 ' (1-20 of SEQ ID No.2)；

R：5 '-taagacggacgattaaagtt-3 ' (the 11802-11821 reverse complemental sequences of SEQ ID No.2 Row).

The complete genome DNA of African wild rice W1411 is extracted using CTAB methods.Using this DNA as template, amplification length is about The DNA fragmentation of 12kb, sequence are SEQ ID No.2.

1-2208 of SEQ ID No.2 are the endogenous SH3 gene promoter sequences of African wild rice W1411, the 2209-11122 for containing introne SH3 gene orders (2284-6027,6151-7505,7633-10168, 10218-10378,10455-11014 are intron sequences).The corresponding SH3 gene C DS sequences without introne are such as Shown in SEQ ID No.3.

Two, the structure of expression vector

Expression vector is specifically by Wuhan Biorun Bio-Tech. Co., Ltd. (http://www.biorun.net/) structure, item Mesh number is MOLE20161879.Specially selected in the SH3 gene C DS sequences (SEQ ID No.3) of African wild rice W1411 Target is selected, the present invention is rice U3 promoters using snoRNA promoters, and starting transcription site is A (U6 G), generally The target spot of CRISPR/Cas9 is selected as 23bp length, therefore target spot need to meet AN20GG, considers further that the spacer of 20bp adds its gRNA The RNA secondary structures of skeleton, the present invention choose following four target spots, and four target sequences are specially：First target sequence： AGCAATAGCATGCTAAACA, specially 6040-6058 of SEQ ID NO.2；Second target sequence： ACATCGTGACCGTCCGTTG, specially 6056-6074 of SEQ ID NO.2；Third target sequence： TCCGTTGTGGCCATTGCAC, specially 6068-6086 of SEQ ID NO.2；4th target sequence： TCAGTACTGCAGCAAAGAA, specially 10435-10453 of SEQ ID No.2.It is then respectively synthesized following 3 pairs again Primer, the enzyme enzyme site Bsa I (GGTCTC) in primer front end are expanded (its using sgRNA skeleton+OsU3 promoters as template Shown in particular sequence SEQ ID No.4, wherein 1-76 are sgRNA skeletons, 77-472 are OsU3 promoters).

SH3-1F：5’-cagtGGTCTCaggcatgtttagcatgctattgctgttttagagctagaaatagca-3’

SH3-1R：5’-cgatGGTCTCatcacgatgttgccacggatcatctgcacaactcttttaaa-3’

SH3-2F：5’-cagtGGTCTCagtgaccgtccgttggttttagagctagaaatagcaagttaaaat-3’

SH3-2R：5’-cgatGGTCTCaccattgcactgccacggatcatctgcacaactcttttaaa-3’

SH3-3F：5’-cagtGGTCTCaatggccacaacggagttttagagctagaaatagcaagttaaaat-3’

SH3-3R：5’-cagtGGTCTCaaaactcagtactgcagcaaagaatgccacggatcatctgcacaa-3’

Wherein, first target sequence corresponds to 16-34 (reverse complementals) of SH3-1F, and second target sequence corresponds to SH3- 16-20 (reverse complementals) of 1R and 12-25 of SH3-2F, third target sequence corresponds to the 17-20 of SH3-2R 11-25 (reverse complementals) of position and SH3-3F, the 4th target sequence correspond to 16-34 of SH3-3R.

Above 3 kinds of amplified productions Bsa I digestions are recycled respectively, with T4 ligases and Bsa I after 3 kinds of segments are mixed Carrier pBWA (V) H-cas9i2 skeleton large fragments of digestion connect, and obtain African wild rice CRISPR-Cas9 gene editings and carry Body is named as CRISPR-Cas9-SH3 after sequencing is correct.

CRISPR-Cas9-SH3 carrier structures are described as：CRISPR-Cas9-SH3 carriers are in pBWA (V) H-cas9i2 The Bsa I restriction enzyme sites of empty carrier insert the recombinant plasmid obtained after sgRNA expression cassettes.Wherein, the sgRNA expression cassettes of insertion Particular sequence as shown in SEQ ID No.5, which is followed successively by " SH3-1F+gRNA+OsU3 promoters+SH3-1R+SH3-2F + gRNA+OsU3 promoter+SH3-2R+SH3-3F+gRNA+OsU3 promoters+SH3-3R ".Wherein, 1-45 are SH3-1F Primer sequence (does not include restriction enzyme site and protection base), and 46-100 are gRNA sequences, and 101-496 are opened for OsU3 Promoter sequences, 466-505 are SH3-1R primer sequences (reverse complemental) (not including restriction enzyme site and protection base), the 502-545 are SH3-2F primer sequences (not including restriction enzyme site and protection base), and 537-591 are gRNA sequences, 592-987 are OsU3 promoter sequences, and 957-996 (do not include digestion for SH3-2R primer sequences (reverse complemental) Site and protection base), 992-1036 are SH3-3F primer sequences (not including restriction enzyme site and protection base), the 1028-1082 are gRNA sequences, and 1083-1478 are OsU3 promoter sequences, and 1458-1502 are SH3-3R primers Sequence (reverse complemental) (does not include restriction enzyme site and protection base).

Three, Africa wild rice W1411 and identification are converted

1, convert and be sequenced identification

By the CRISPR-Cas9-SH3 of step 1 structure by Bombardment-Mediated Transformation into African wild rice W1411, with containing tide The NB culture mediums of mycin carry out 2 wheel screenings, then through pre- differentiation, and differentiation obtains transfer-gen plant.It tests while being arranged to African wild The control of pBWA (V) H-cas9i2 empty carriers is transferred in raw rice W1411.

Transfer-gen plant is identified by sequencing.Concrete operations are as follows：Growth period transformed plant and the blade of W1411 are taken, Total DNA is extracted using CTAB methods, sequencing identification is carried out for the above plant using following primer：

SH1-exon2F：5’-CGGCTTACTTGTATTTCTTCCA-3’；

SH1-exon2R：5’-GCATGGTGTCTTTTCCGTTT-3’.

As shown in figure 3, W1411 is the acceptor material of non-transgenosis, Cas9-1, Cas9-2, Cas9-3 distinguish qualification result For 3 independent transgenic lines.As seen from Figure 3, having respectively obtained the sites SH3 gene C DS has a 1-bp insertions, 5-bp missings with And the transgenic line of 19-bp missings, as a result lead to the variation of its encoding histone, so as to cause the reduction of its seed holding.

Four, the Function Identification of transgenosis rice

Identification is screened and be sequenced through step 2 hygromycin resistance obtains 9 plants of transfer-gen plants altogether.It is planted with this 9 plants of transgenosis Strain, and the W1411 for being transferred to the control of pBWA (V) H-cas9i2 empty carriers and being handled without transgenosis is experiment material.Observation The seed holding of each experiment material.

As a result it shows：

Compared with unloaded control and the W1411 handled without transgenosis, screens and be sequenced through step 2 hygromycin resistance and reflect Fixed transfer-gen plant is more difficult to shattering, Cas9-1, Cas9-2, and Cas9-3 is respectively 3 independent transgenic lines (Fig. 4).

<110>China Agricultural University

<120>With the relevant albumen of rice seed holding and its encoding gene and application

<130> GNCLN180982

<160> 5

<170> PatentIn version 3.5

<210> 1

<211> 186

<212> PRT

<213> Oryza barthii L.

<400> 1

Met Ser Ala Gln Ile Val Pro Ala Pro Glu His Val Cys Tyr Val His

1 5 10 15

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

20 25 30

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

35 40 45

Ser Val Asn Leu Arg Gly Leu Val Gln Ala Phe Pro Ala Glu Asp His

50 55 60

Leu Gln Asp Asn Leu Lys Met His Asn Met Ser Phe Arg Glu Asn Tyr

65 70 75 80

Ser Glu Tyr Gly Ser Ser Ser Arg Tyr Gly Arg Val Pro Met Met Phe

85 90 95

Ser Lys Asn Asp Thr Glu His Met Leu His Val Arg Pro Pro Glu Lys

100 105 110

Arg Gln Arg Val Pro Ser Ala Tyr Asn Arg Phe Ile Lys Glu Glu Ile

115 120 125

Arg Arg Ile Lys Ala Asn Asn Pro Asp Ile Ser His Arg Glu Ala Phe

130 135 140

Ser Thr Ala Ala Lys Asn Trp Ala His Phe Pro Asn Ile His Phe Gly

145 150 155 160

Leu Gly Ser His Glu Ser Ser Lys Lys Leu Asp Glu Ala Ile Gly Ala

165 170 175

Pro Ser Pro Gln Lys Val Gln Arg Leu Tyr

180 185

<210> 2

<211> 11821

<212> DNA

<213> Oryza barthii L.

<400> 2

tgcagagact tccgggttga cggatgccac ctctgcacgt aaaaactgtt tttgcccgca 60

caggaaaatt gttttcgtag cagtgaaaag aggtgtaaaa aagcacgcaa cgcatgcata 120

tcaaataatt cgtcgaatac ggaacaaatt ggaatcgcta gctatagtct ctgttcacca 180

ccaaaaagaa tcccacaaaa aaataattct taacccttct tgtaggagta ctccatatat 240

atacgtaatg ccgtggctta atttactatt gcaccgtaat cttgtgatca aggacttccc 300

cttatttggt taattaggtg aagcaccaac tgactaaaag tgtatatata gttgcgtatt 360

tagtcgtccc acaaactttc caattttcca agatgcgggt gcatatatat atatatatat 420

atatatatat atatatatat atatatatat agtgtaaatc aacttcgtaa ttaagcataa 480

ctgtccactt gtactttctg caaaatgtga tacatcctct gtagcaggtg tggaccaata 540

agaaccacac tatcaaaagc agttctgaaa atatatttat tccccctctt tccaaaaaaa 600

ataaaacagt ggatgaatca tgggcgcaac atggtaataa tataaaaatg attgttaaca 660

cataataatt ccaacaatgt gaagaacaat tttgtcttta acaacttttt tttcttttta 720

aaaaagcaca tgtttagcga tttatttcta gaaaccccct aaagggtatg ttcggaactg 780

ctggttccca gcttctccgc ctccttttcc gcacgcacgt ttttcaaact gttaaacggt 840

gcgtttttta ccaaaagttt atatacgaaa gttgcttaaa aaatcatatt gatctatttt 900

ttttaaaaaa taggtaatac ttaattaatc acgcgctaat agaccgctcc gttttccgtg 960

cgtgtgagat gggttcccaa cacccacgaa caaacacagc ctaagagttc ggcaaaaatt 1020

tcttctgttg aacttcatga aaaaaaaatg tttcctacaa aaggtcattg cactacaaat 1080

aaagtgagat gacagaagct tgatgtttca gatagctgat ttagtcaaat atttaagcgg 1140

ttcggttgta cgtccaacct ctacctgaat taaaattcta gatttagatc cagcacccat 1200

caatttgata agtacacaca tgcttgtcca tactcgtgcc cgcgggtgtt tgctgccttt 1260

gataatatcc cagtttgcgt agagaataca cacacgtata tgggttagtg tgtgcatcaa 1320

atgcattctc ttaaaaagaa agaaataatt gagatatttt aaaaagccta aggtacattt 1380

cttgtctaaa ttaataccat tatgaataat atatttgtat tcgaaaccaa cattcgcacg 1440

aagtatcctt acgaaaaatg tttgaaacct gaaatttctc agaatgatat aatagttaaa 1500

atataatatg gcagcttctc aacatattat tcaaagaacc actacctcaa gataacttga 1560

attttaatat tagatgagtt atatagaaag accagtttgt gttgacatga catgtggctg 1620

ctgagtagtt acatgcagac aatccaatcc atcctttggg ggtacaatga gggagccccc 1680

aaccattcat tgttgcaatc atgctagcta ctatagcatt ttccacctcc aacaccaccc 1740

ccttcacatg agccacgcaa caaaaaggcc tattcccccc tcaccctctc ccgtttgcac 1800

caccaccaat ctctctctct ctctctctct ctctctctaa aagatctcta tctatctaaa 1860

gatctgagct acaattaatt aaaacctgat aagaagaaga ccctgagctc aatcccttct 1920

tattatacta tatatatact tctgtatttg tgcacagtgt tggtacaggt atctgcagac 1980

agccatatct atctttctag ctaattggca atatgcctga agtccactgc tcgtcctaag 2040

atttacttct ccaccacccc cccccccccc ccactccttt gcttcttctt cttcatcatc 2100

atcttcttcc tcctcctgtt aattaagttc ttgatttcta agctagcaat tgagcacaca 2160

tcgatcgcga gagacgcagc ggcgcccgtc taccgcgggg ctagaggaat gtcggcacag 2220

atcgtgccgg cgccggagca tgtgtgctac gtgcactgca acttctgcaa cacaattctc 2280

gcggtaattt gtctgaattg ttgatccctt tctccttctc tctctctctc tccccttcct 2340

tgatttcctt gcctcctctt ttctacatga agtttctttt cttttttggg ggggaactct 2400

cgatgaaggt gcagatctac aagggcaagg gagatcgatg catatatgag atgattggtt 2460

gcttggcttt tctttagaat tgtgtatgat ttgggcagtt catttgcgtg atttctttaa 2520

tttttaattt gtgcgaaaag atgtgtgtca tgctgccaag agattgggtt tcgatatata 2580

tgttactttt tttttaccca attacccttc gttttctttc tttaattgtc gacagagttc 2640

agatagcttc acctcctttc cttttcgact cattaaaact ctggtccggt tttctctgaa 2700

agagagcttg tcgaccctaa tttttttgat ttagtctatc atggagtcat gggcatgatt 2760

acatatgaat gggaaaaagg gcaaaggtca tgatgatctt gagaattgag ctgcttctct 2820

ctctttgcac cattttgtgt cgtttgatta tatgggttca tgaattattt gggcgagttg 2880

atgaacttca attaactggt acgtactaga gtagctagct ctagatcatc cttgttttct 2940

ctctcgactc ttaatttcct tcagaaccta gcgtagctga tgctagatcg atcttacgta 3000

catatgaagt gcagttgatt ttcttgccgg tcgactggta ctttatatgg ctaccatata 3060

taagagtatc gtcgtttcag gtgttattac tagtaatcga attgctacaa attaaagagc 3120

atgcaataat aacacataac tttaattagt atatgcaact actgtgtgta atgttctcct 3180

gcttgcctct tagatcgatc atcttttaaa atccacttgc tcatcccctc taagcacaaa 3240

tacagcatct gaaggactca gaaacggcaa cctctcgaag tacggtactg tgttacaaga 3300

attaaggtgc aaattgcatc aataattcct gtatgtatgt gttcccgtgc tagctgttcc 3360

actcattcgt tcttcccgtg ctatatcttg gctttcttga tggtttctct ttcttttttt 3420

tcttttcttt ttggtgcaat gcatggtgtg agtttaagga ctaagtacaa ataactaaaa 3480

ctacataatt aaaatgaagc agaagcagct agtaaccata aggcaaaaag attgttacgt 3540

ccttccaaag aaaaaaaatt gtggtttgag cttgagctag cttcttctcc ttcctcctct 3600

tctttttttc cagagagaga gagagagaga gagagagaga gatgcttgag atggcatggc 3660

cggctagctt cactcaatac atgcaaggga attagatcgg tccttatata tagactatag 3720

tcatcagtga tgtgttgaac tggtctgaat ggcgttttaa ctagctaatc cccatgttga 3780

actcatatat acactatcag gaattcgtat aaacatttta atttctctct ctattttttt 3840

ttctgttttc ccacttcttc ttggatcttg ccttgcttgc tttggaagta cacgctgcta 3900

gactagtgat atgatatgat cgatccccct cagtcagtca gtctgatctg catatatggg 3960

cggcgagagt gctacgaaat atcgcaaatt cgtgcacaca cacaacataa caaatctcta 4020

gctagttttc tacggttctc aatgaattga agtatatata tctttatata tagtgcatat 4080

caattattct tgttgattaa ttagttagtt ccgttcgatt ctcgacatgt taacttcttg 4140

gcacaattac atgcatacag tctttatata ggagtatata tatcttgttg atctgcgcta 4200

cttgcatgct tcaatatatc tttcagactt tcagaagaga aattatattc cttatctctg 4260

aagtctaaat agctactgta gttcgcaaga actttattgg aggccatctt ttactccatc 4320

tcaatctctc ctactccctc cagaaagata atcaaaataa gcgcatttgg gaacaaatat 4380

agctagtgac cattaatgag agtcgggccc aaacaattca agtattgtaa ataccataac 4440

agcaaaaagt agaatacacc aaaatgagtc tattaccagt tccaggaaga gcacaccaaa 4500

tgacagaaac tattactcaa tcccaaaaca aattaatttt tactatagct aattagctat 4560

gaacttggaa agtagtgacc gggatataga gagtgtacac gccaacgaca aatgcagagc 4620

cgtacgtact gacttcctgc atctacatca cccactgtaa tgtcaattgc tgaggcgtag 4680

tacacatgtc tagtgtagat agtacatgta tagctacata tgcatccatg catgatcttt 4740

cacaagatct taccagcatg tctcaacttt actactgcaa ctccaatact tatatatgta 4800

tgtcttcttg ctctctccta ccgaaagatg aaaaaaaggg gattgaatca tcttgtgcat 4860

tgcatgaccc acacttaaat gtactccctc catccaaaaa aaactcaacc acccctccta 4920

agacaacacc cctcctaaga caacgaatct gaacaaaagg tagtctaaat tcattgtcca 4980

catcctcttt taggttgagt tttttttgga tggagggagt acccacacaa acacacatgc 5040

aggagaggag atgttgtgct tgagagtgaa ttggcttctg catgaagggt cactgtctct 5100

agctagcacc tttctttcct gcactgttca tgcatgtagc tgaaggcttt gtgatttcct 5160

actactgcta ctactactct gtggagaggc cagacagaga ggcatgacat gacataccat 5220

gcaaatgcat ggcttcttcc tttaaaaaag tcaccaatag atatgttgtg aggttgtttg 5280

cttctgtcac aaccaactac tagcagctag cccttttgtg ctagatatgt gtggagtgaa 5340

gtgtgatttg agctaactcc cctcatggga gcacacacaa tcattctcat tcagtcactt 5400

tgcttaatta tttctttctt ctttcttatg ggagtgagtg cccaaaaagt gagtgaatgt 5460

agggaatcaa acaaagtaga ggtactgtgc tcttcaggtt tgggaagaaa aatgggaacg 5520

gcaagggacc aatgtgtgca ggcattggaa aagggtgccc caattctgat ctgatcagat 5580

atatattcag atcagcagag cccctgataa aagtcatgga ggaacagtta tccacatcta 5640

aagctatagc tgtgtattaa tgtgtatctg cagtctacac cagccttatg aaacatgcat 5700

gcatgcatgc atgcctttga gatttgattt tttacttctc agattctttt gtacaaatgc 5760

aaaaagaaaa tgatcctttg gaatctagca tttgttacat atttacaata taaagtgcta 5820

tacttgttta cttttcattc tacttgttcc agcacatgca tatgtaattg tacacctgct 5880

caaatttaca atgagtggca tatatattat acatattaat tacggcttac ttgtatttct 5940

tccataaaag aacatgtcta gaatatttct tttcttgaca gtaaatattc aggacatatt 6000

atactgattc ttggaatcaa tctacaggtc agtgttccaa gcaatagcat gctaaacatc 6060

gtgaccgtcc gttgtggcca ttgcactagc ctgttgtcag tgaacttgcg aggattggtc 6120

caagcattcc cagcagaaga tcatttacag gttgacacat caatatatac tactctttca 6180

tgcatgaaat tatggataca gatacaatga ctacattaaa tcatattatc tctttttaat 6240

aaaaaaacac acatatgttg cctatatata tctagaatcc aaattaatgg cacttttcaa 6300

ttataaccat tgctactgca tgcttagtct gatagagcta gaaatcaacg aacaattaat 6360

gtatatgtac cggagcattc ttcacatacc cagctggaaa aaaatcacat gcagatcgat 6420

gtatgttaaa ctaccctata tatgtagaag tagtatagta ctactagtat atgtagataa 6480

ctactccctc cgtttcacaa tgtaagtcat tctagcattt cccacattta tactgatatt 6540

aataaatcta gacatatata tctatctaga ttcattaaca ttaatataaa tgtgagaaat 6600

gctagaatga cttatattgt gaaacggagg aagtagattt tactcacaga gaaaaacaga 6660

aaagacacca tgcaatgcat gcatgggtca tctcaagctt gaatacttta catgtacagc 6720

agagacaaat taaagcattg cactatttac tcacaggaaa gaattttaaa gctcttgcac 6780

atgccaaaat atatactagg agtaccaagc catatggcta gatcagctag gtagctcagg 6840

ttgctcagct gatcttatac tacatgactt gtgccccttt gtcaaaccct gcagggttat 6900

tccccgcaaa aaaaaaaatc ctgtagggtt attgataaat actagtagat gttttgcatt 6960

tgtttttata tgctactata tataatggaa aaaatcctgc cacacaagta cttgaattaa 7020

atttccaata tgaaaagcac gagtttcgaa ttccggcaat aactacatgt taggtttgat 7080

atactccctc cgtcagtaag gtgtgtttag ttcacgctaa aattggaagt ttgattgaaa 7140

ttgaaaagat gtgacggaaa agatggaagt ttgtgtgtgt aggaaatttt tgatgtgatg 7200

gaaaagttgg aagtttgaaa aaaaagtttg gaactaaact cggcctaaat atgagggata 7260

cgttactgta gaatgtacaa aattgattcg ccaaattcat gtttagagat tttcatgttt 7320

gcttcacaaa acaatcaaaa gccatgcaga tttttcgaaa tgatctgaac agtaaccacc 7380

aatgttttac ttaattttct tgtcttgact tggttataat ttccatgtaa atacacacta 7440

agtagcatct agaggaaaaa tgtgtgactt taagttgatt aacttgcttc ttacattttc 7500

ctcaggacaa tttgaagatg cacaatatga gcttccgcga aaactactct gaatacggct 7560

catcttccag atatggccga gtgccaatga tgttctcaaa aaatgacaca gagcatatgc 7620

tgcacgtgcg acgtaagaca atgcatatga acctgcataa taaatttagt gtacatgtgc 7680

gacggagcca tatttctaga gattattgat tttactatat gaagcaatat aaaggtatac 7740

aaatgctagg ggaatattgc atatgttcac atatacatgc tgggtgcata taccactttt 7800

cttctctaaa ccctttttat ctttgttgtt ttgctcagac aactggggta cgtgtgttac 7860

ttcttaatta acacaaatac atgtatagag aaattatact ggcttaatca attcctgagt 7920

tatgcacttt tttaaacaga tatatttaga aaattcgacg agggataaac ctctactaaa 7980

aagtcagaaa aatagaaaat ttaaacaagt tgcattgatt cagctagagg tggataataa 8040

actagctagg ctcacaagct ggcttgttaa gctcgagttt tatatcaatt taacatgtat 8100

taatctatta tatgttcaca taatagtaaa cactactccc tctgtttcat attaaagtcg 8160

tttcatatta taagttattt tgattttata aaaaaaagtc gttttgattg ttttcctagc 8220

tagtcaaagt aatttaagta tgaccaagtt tacagaaaaa tataacattt ctaatacaga 8280

acaaacatat taaaacatat tcaatgctag atttaatata actaatttgg tgttttaagt 8340

gttgcaaaag gtttctataa atttaatcaa aactttaaaa attttgacta gaaaaagtca 8400

aaatgactta ttatgtgaaa cggaggaaga gtaactagta aagtactaac tagacatgaa 8460

tagtttgaaa gtaatttcaa gaagtattat ccatagtata agtattgact aataggcact 8520

cggtatatat ggaaaatatg gtgtgtctag ttaagctaac aagctaaaaa tccatctcac 8580

aagttcgagt cgatcagcca agccacaagc tttgagtttt tttccccaac cctagctagg 8640

cttagtaaag cttgttccac ccaagatgcg cgtctgctgt tattgatcac gttaagttct 8700

gttaaagttt atggtcccca tcgcttggct tatttcctaa taagccaaaa cggcttatta 8760

gaaaataaaa atgaatttgt aggcaaaact tttataattg tgttcttggt gacttaaaag 8820

ccaatgttga aaagaaacta cgttgaaaat atctcaaaat caatctcaaa attaagcttg 8880

aaaatttaaa atttggcttt ttctttggtt gattaggcca tccgatggga gcctatactg 8940

ggctgtatat agaaattatt cttatatgaa ttaatgaatg aaacgagatg tgcatgtgtg 9000

cgtgagttct agctagtagc tagtaccatg catggttcgg ttaaagattt ttccgtgcgg 9060

tcaaaataca tttttcagtg cggaggtccc acccgtctgt accctgcatg tctgcgaaaa 9120

tccatatttt cgcgtgcggg cggcctaacc gcatgagata atccatcttc ccgtgcggtc 9180

gggttaagta gaccacacgg gataatcgat tatcccatgc ggtcaaatta taccgcccgc 9240

acgcaaaaaa aaactgaaaa aaaataaaat caaaaacact aaccctaacc ctagaaatct 9300

agaatcgccc gctgccgctg ctcgtgttcg ccatcgtcct catgtcgtcg tcgtggtcgc 9360

gccgtcgccg ctctcatcat cgtcgttgtg gtcgcgctgc cgccgctccc ggccgccgtc 9420

actgctcccg gttgccagcg cggcgcggat ccgaccggtc acccgccgcg ccgtctccgc 9480

cagcagccgt cgtgccgccc gctcccggcc gccgcctcgt tggcggcaag ccgggcaact 9540

cccgcgcccg tgcgctcggt ggggagggga ggagctagag gaaaggcaga gagagagaga 9600

ggagtgattg agggtgaata aaagggaaaa ggaatagaga agatgaaaag tttaaagtga 9660

gtaagagaga aaagagggag aggtattttc gcgtgcagtc cacttaagag gcccgcacgc 9720

gaattcgcgt gcgggcctct taagggtcag cacgcaaaaa taggcttatt tttgcgtgcg 9780

gaccgttaag aggcttgcct gcgaaaatcg atatttgcgt gcaggtctct taagagtccg 9840

cacgcgaaaa tgggaggcca atttttgcag acgcgcgcga aaacggtccg tctgggaaaa 9900

tggaagctcc ttgtccagaa aaatgttttg tgtactagta tatatttgaa aatactgatg 9960

catgaactag ctacttgagt actatttcat tttgtgagcc caaaaatatt gacgtacgaa 10020

ctacttgact actacgtact cctattaatt tcttgaaaaa cctaggagta gtatacatac 10080

tctcaagagt atgtgcttta gatagtaatt aaacagtatc tggatgctta tacttttttt 10140

ttacaaatct ttttcataca acttgcagct ccagagaaga ggcagcgggt tccttcagct 10200

tataacagat ttatcaagtg agtttgatct cgagacaatt cgatcgatcc atatatatat 10260

atatagatga tctagacagc ttataacatc tgagagtttt cagataattg atataaatct 10320

gccgtattct atactaagag agtgtttgat tttgacaccc actaatatat cactacaggg 10380

aagagatccg gaggataaaa gcaaacaatc ctgacataag ccacagagaa gccttcagta 10440

ctgcagcaaa gaatgtgagt acatgtgtag ctaggggaaa caaattacta tcttaaactt 10500

ttacatatac tgatataata gctgttcttt tttttactac aaactataat gcaaattctc 10560

acacactttg gcctggttta gttcccaact ttttcttcaa acttctaact tttccatcac 10620

atcgtcccaa ttttaaccaa acttctaatt ttagcgtgaa ctaaatatac ccttgatcat 10680

tgctgaatat atattgatag aagtatgtga gccatataca tagagttaca tatcaacaat 10740

actacatatt gttgactttg tatgtcactg ctagcaagat attaaatcca tggcaattgc 10800

catttgctta atactcttat acatatagta gtgtccatat tgatcactaa atactcagtc 10860

tatatgaatt taatttatat aacatattcg taaaaaaaat agtggtcaaa gttatatatt 10920

taagaccatg tcatagtcta aaccatcatc tacttctgac tagagggagt aactctcatc 10980

tcatatgcat cctacttttt tttttttttt gcagtgggcg catttcccga acatccattt 11040

cgggttaggc tcccatgaga gcagcaagaa gctcgatgag gccattggag ctcctagtcc 11100

ccaaaaggtt caaaggctgt attgatcgag tacacatgat gatcgatgga tcctacgtgc 11160

ttgttgatct catgttgtaa gtaacttaat taagaaaaaa agtctacgct tatttatttc 11220

ttaagtatca tcagttaaat ttgtgagcga gtatatagat gagttgtcat gatcagtgag 11280

tgatcgtatg catgcagtgc tgagaattaa aggtgtatat gcatgagtaa cgttacatgc 11340

agctagtgtc gacgcacgta tgacagactg aataaactct tcaccatgtt gtttttttat 11400

gtttgaaatg aattaattaa ttgtccatga ttgcaatcca tttcataccg gtattattgg 11460

ccagtgcaga tcaaagtttc tacagcttat aagtactata tattgtatcg tatatgtgtg 11520

cacgatgcac taatccattg tcatttgtct acacggtacg caagatcgat catactgtgt 11580

acacatgcat actgtgcata gtgctgttct tttgtgctaa atactaaaca tatgaattgt 11640

caatcagtaa tgtactacta tattatatat ataatactat atgtatgtag attttaatga 11700

aagattgagc ttgcatttat cctgcaattg agaaacattg gaccaggtga aatggtactc 11760

cttccgttcc attttaagtg caaccataag tttccgtgcc caactttaat cgtccgtctt 11820

a 11821

<210> 3

<211> 561

<212> DNA

<213> Oryza barthii L.

<400> 3

atgtcggcac agatcgtgcc ggcgccggag catgtgtgct acgtgcactg caacttctgc 60

aacacaattc tcgcggtcag tgttccaagc aatagcatgc taaacatcgt gaccgtccgt 120

tgtggccatt gcactagcct gttgtcagtg aacttgcgag gattggtcca agcattccca 180

gcagaagatc atttacagga caatttgaag atgcacaata tgagcttccg cgaaaactac 240

tctgaatacg gctcatcttc cagatatggc cgagtgccaa tgatgttctc aaaaaatgac 300

acagagcata tgctgcacgt gcgacctcca gagaagaggc agcgggttcc ttcagcttat 360

aacagattta tcaaggaaga gatccggagg ataaaagcaa acaatcctga cataagccac 420

agagaagcct tcagtactgc agcaaagaat tgggcgcatt tcccgaacat ccatttcggg 480

ttaggctccc atgagagcag caagaagctc gatgaggcca ttggagctcc tagtccccaa 540

aaggttcaaa ggctgtattg a 561

<210> 4

<211> 472

<212> DNA

<213> Artificial sequence

<400> 4

gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt 60

ggcaccgagt cggtgctttt tttgtcgtag aaaggaatct ttaaacatac gaacagatca 120

cttaaagttc ttctgaagca acttaaagtt atcaggcatg catggatctt ggaggaatca 180

gatgtgcagt cagggaccat agcacaagac aggcgtcttc tactggtgct accagcaaat 240

gctggaagcc gggaacactg ggtacgttgg aaaccacgtg atgtgaagaa gtaagataaa 300

ctgtaggaga aaagcatttc gtagtgggcc atgaagcctt tcaggacatg tattgcagta 360

tgggccggcc cattacgcaa ttggacgaca acaaagacta gtattagtac cacctcggct 420

atccacatag atcaaagctg atttaaaaga gttgtgcaga tgatccgtgg ca 472

<210> 5

<211> 1502

<212> DNA

<213> Artificial sequence

<400> 5

aggcatgttt agcatgctat tgctgtttta gagctagaaa tagcaagtta aaataaggct 60

agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgtc gtagaaagga 120

atctttaaac atacgaacag atcacttaaa gttcttctga agcaacttaa agttatcagg 180

catgcatgga tcttggagga atcagatgtg cagtcaggga ccatagcaca agacaggcgt 240

cttctactgg tgctaccagc aaatgctgga agccgggaac actgggtacg ttggaaacca 300

cgtgatgtga agaagtaaga taaactgtag gagaaaagca tttcgtagtg ggccatgaag 360

cctttcagga catgtattgc agtatgggcc ggcccattac gcaattggac gacaacaaag 420

actagtatta gtaccacctc ggctatccac atagatcaaa gctgatttaa aagagttgtg 480

cagatgatcc gtggcaacat cgtgaccgtc cgttggtttt agagctagaa atagcaagtt 540

aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttgt 600

cgtagaaagg aatctttaaa catacgaaca gatcacttaa agttcttctg aagcaactta 660

aagttatcag gcatgcatgg atcttggagg aatcagatgt gcagtcaggg accatagcac 720

aagacaggcg tcttctactg gtgctaccag caaatgctgg aagccgggaa cactgggtac 780

gttggaaacc acgtgatgtg aagaagtaag ataaactgta ggagaaaagc atttcgtagt 840

gggccatgaa gcctttcagg acatgtattg cagtatgggc cggcccatta cgcaattgga 900

cgacaacaaa gactagtatt agtaccacct cggctatcca catagatcaa agctgattta 960

aaagagttgt gcagatgatc cgtggcagtg caatggccac aacggagttt tagagctaga 1020

aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt 1080

gctttttttg tcgtagaaag gaatctttaa acatacgaac agatcactta aagttcttct 1140

gaagcaactt aaagttatca ggcatgcatg gatcttggag gaatcagatg tgcagtcagg 1200

gaccatagca caagacaggc gtcttctact ggtgctacca gcaaatgctg gaagccggga 1260

acactgggta cgttggaaac cacgtgatgt gaagaagtaa gataaactgt aggagaaaag 1320

catttcgtag tgggccatga agcctttcag gacatgtatt gcagtatggg ccggcccatt 1380

acgcaattgg acgacaacaa agactagtat tagtaccacc tcggctatcc acatagatca 1440

aagctgattt aaaagagttg tgcagatgat ccgtggcatt ctttgctgca gtactgagtt 1500

tt 1502

Claims (10)

1. protein is following any：

A) protein that amino acid sequence forms shown in SEQ ID No.1；

B) amino acid sequence defined by a) by the substitution of one or several amino acid residues and/or is lacked and ored add, And with the relevant protein of the seed holding of vegetable seeds；

C) and a)-b) in it is any defined by amino acid sequence have 95% or more, 90% or more, 85% or more or 80% with Upper homology, and with the relevant protein of the seed holding of vegetable seeds.

2. encoding the nucleic acid molecules of protein described in claim 1.

3. nucleic acid molecules according to claim 2, it is characterised in that：The nucleic acid molecules are described in coding claim 1 The gene of protein, the gene are following any DNA molecular：

1) DNA molecular shown in 2209-11125 of SEQ ID No.2；

2) DNA molecular shown in SEQ ID No.3；

3) under strict conditions with 1) -2) in the DNA molecular of any restriction hybridize and encode protein described in claim 1 DNA molecular；

4) with 1) -3) in any restriction DNA sequence dna have it is 95% or more, 90% or more, 85% or more or 80% or more same Source property, and encode the DNA molecular of protein described in claim 1.

4. the recombinant vector, expression cassette, transgenic cell line containing nucleic acid molecules described in Claims 2 or 3 or recombinant bacterium.

5. recombinant vector according to claim 4, it is characterised in that：The recombinant vector is recombinant expression carrier or recombination Cloning vector.

6. the recombination described in nucleic acid molecules or claim 4 or 5 described in protein or Claims 2 or 3 described in claim 1 carries The application of body, expression cassette, transgenic cell line or recombinant bacterium in the seed holding of regulation and control vegetable seeds.

7. method, for following (A) or (B)：

(A) a kind of method reducing plant fallen, includes the following steps：Reduce albumen described in claim 1 in recipient plant The expression quantity and/or activity of matter；

(B) a kind of method improving plant fallen, includes the following steps：Improve albumen described in claim 1 in recipient plant The expression quantity and/or activity of matter.

8. method, for following (C) or (D)：

(C) a kind of method for cultivating the genetically modified plants that seed holding reduces includes the steps that following (a1) and (a2)：

(a1) encoding gene of protein described in claim 1 in recipient plant is knocked out or inhibits to express, obtain turning base Because of plant；

(a2) it is obtained compared with the recipient plant from genetically modified plants obtained by step (a1), the transgenosis that seed holding reduces is planted Object；

(D) a kind of method for cultivating the genetically modified plants that seed holding improves includes the steps that following (b1) and (b2)：

(b1) encoding gene that protein described in claim 1 is imported into recipient plant, obtains expressing the encoding gene Genetically modified plants；

(b2) it is obtained compared with the recipient plant from genetically modified plants obtained by step (b1), the transgenosis that seed holding improves is planted Object.

9. according to the method described in claim 8, it is characterized in that：It is described " to claim 1 in recipient plant in method (C) The encoding gene of the protein is knocked out or inhibits to express " it is realized by CRISPR-Cas9 technologies；

In method (D), " encoding gene that protein described in claim 1 is imported into recipient plant " is by described Import what the recombinant expression carrier containing the encoding gene was realized in recipient plant.

10. according to any application or method in claim 6-9, it is characterised in that：The plant is monocotyledon Or dicotyledon；

Further, the monocotyledon is grass；

Further, the grass is rice.