CN109295071A - A rice flower organ development regulatory gene PEH1 and its encoded protein and application - Google Patents

Abstract

The present invention provides protein and the application of a kind of rice flower organ developmental regulation gene PEH1 and its coding, and the gene PEH1 has the nucleotide sequence as shown in SEQ ID No:1；Addition, substitution, insertion or deletion one or more nucleotide in the SEQ ID No:1 nucleotide sequence and nucleotide sequence, allele or the derivative generated；The protein has the amino acid sequence as shown in SEQ ID No:2, addition, substitution, insertion or deletion one or more amino acid in the SEQ ID No:2 amino acid sequence and the amino acid sequence or derivative that generate；The development of floral organs that the protein of the gene PEH1 and its coding is applied to rice is adjusted.The present invention is regulated and controled using gene pairs rice flower organ development, has important theory and practical significance to further investigate the molecular mechanism of rice reproductive development.

Description

Protein and the application of a kind of rice flower organ developmental regulation gene PEH1 and its coding

[technical field]

The present invention relates to plant genetic engineering fields, and in particular to a kind of rice flower organ developmental regulation gene PEH1 and its The protein of coding and application.

[background technique]

Rice is not only the model plant of monocot genes group research, and is important cereal crops.Rice is The staple food of the one third mankind in the world, flower are not only organ of multiplication, and are the bases to form seed.Rice is spent normal Development directly affects rice yield and rice quality, therefore the research of rice flower development is always that Genetic and breeding in rice worker is closed The emphasis of note.Study On Rice flower development not only has important theory significance, but also has to yield and quality of rice genetic breeding There is important directive significance.

In the 1990s, plant developmental biology man is ground by the homeotic mutant to arabidopsis, the goldfish showy flowers of herbaceous plants The floral organ for studying carefully discovery dicotyledon is formed by controlled by multiple genes, associated with each other between gene, proposes there is milestone significance ABC model (Coen ES and Meyerowitz EM.The war of the whorls:genetic Interactions controlling flower development.Nature, 1991,353 (5): 31~37.).With The increase for the floral homeotic genes quantity of research goed deep into and cloned, ABC model extend to ABCDE model (Theissen G.,Development of floral organ identity:stories from the MADS house.Curr Opin Plant Biol.,2001,4(1):75-85.)。

It finds to obtain in the research of the species such as dicotyledonous model plant arabidopsis, toad's-mouth by the research of rice molecular science of heredity " the ABCDE floral organ regulation-control model " obtained partially is suitable for explaining the molecular regulation of the monocotyledons development of floral organs such as rice (Yoshida H and Nagato Y.Flower development in rice.2011,J Exp Bot.,62(14): 4719–4730；Lombardo F and Yoshida H.Interpreting lemma and palea homologies:a point of view from rice floral mutants.Front Plant Sci,2015,61(6):1-6.).At present Cloned it is multiple with dicotyledon arabidopsis, the corresponding ABCDE model gene function of toad's-mouth is similar, product is homologous MADS-box gene (in new, Wang Jianjun, the molecule mechanism Molecular Plant Breeding of king's ability woods rice flower organ development, 2013,11 (4): 617-624. it), however is differed farther out in the evolution such as rice and arabidopsis, colored morphosis differs greatly, rice flower tool There are stamen similar with dicotyledon and gynoecium organ of multiplication, but without calyx and flower possessed by apparent dicotyledon Valve structure is bright with dicotyledon and other monocotyledonous calyx, petal around stamen and gynoecium breeding floral organ Show different lemma, glumelle and lodicule.

With the separation of the deep and new rice flower development related mutants gene of research, more and more researchs are aobvious Show, unifacial leaf rice and dicotyledonous mouseearcress flower organ morphology are built up more different still to be needed to solve there are many problem (Hitoshi Y and Yasuo N.Flower development in rice.J Exp Bot,2011,62(14):4719- 4730；Lombardo F and Yoshida H.Interpreting lemma and palea homologies:a point of view from rice floral mutants.Front Plant Sci,2015,61(6):1-6.).Firstly, inside and outside Bran whether be calyx homologue, there is no sufficient experimental evidences for this point to support；Secondly, inside and outside bran is same device Official is different organ and does not obtain ample evidence yet；Third, actually or rice lepicena one degeneration flower one it is whole Piece colored organic component, is still not very clear.Thus, still cannot to the genetic regulation mechanism of the flower development process of rice A general outline is formed as arabidopsis, to illustrate some basic problems of rice flower development, need some new bases Because cloning and studying.

[summary of the invention]

One of the technical problem to be solved in the present invention, be to provide a kind of rice flower organ developmental regulation gene PEH1 and its Using, using the gene pairs rice flower organ development regulated and controled, for further investigate rice reproductive development molecular mechanism With important theory and practical significance.

The present invention is realized in one of above-mentioned technical problem:

A kind of rice flower organ developmental regulation gene PEH1, the gene PEH1 have the core as shown in SEQ ID No:1 Nucleotide sequence.

Further, addition, substitution, insertion or deletion one or more core in the SEQ ID No:1 nucleotide sequence Thuja acid and nucleotide sequence, allele or the derivative generated.

Further, plasmid, plant expression vector, place containing a kind of rice flower organ developmental regulation gene PEH1 Chief cell.

Further, addition, substitution, insertion or deletion one or more core in the gene PEH1 and its nucleotide sequence Thuja acid and development of floral organs that the nucleotide sequence, allele or the derivative that generate are applied to rice is adjusted.

Further, plasmid, plant expression vector, Su Zhuxi containing a kind of rice flower organ developmental regulation gene PEH1 The development of floral organs that born of the same parents are applied to rice is adjusted.

The second technical problem to be solved by the present invention is to provide a kind of rice flower organ developmental regulation gene PEH1 coding Protein and its application, using the protein to rice flower organ development regulate and control, for further investigate rice reproductive organs The molecular mechanism of development has important theory and practical significance.

The present invention is realized in the twos' of above-mentioned technical problem:

A kind of protein of rice flower organ developmental regulation gene PEH1 coding, the protein have such as SEQ ID No: Amino acid sequence shown in 2.

Further, addition, substitution, insertion or deletion one or more ammonia in the SEQ ID No:2 amino acid sequence The amino acid sequence or derivative that base is sour and generates.

Further, addition, substitution, insertion or deletion one or more amino in the protein and its amino acid sequence Sour and generation amino acid sequence or derivative, the development of floral organs applied to rice are adjusted.

The present invention has the advantage that

Gene or protein of the present invention are for rice flower organ (flower glume, male and female organ etc.) normal development to pass Important, the gene or protein mutant cause rice male and female embryo of silkworms to form exception, and flower glume piece form deformity forms capsicum shape Grain husk flower, can be transformed rice flower organ using gene of the present invention or protein, to further investigate rice reproductive development Molecular mechanism have important theory and practical significance.

[Detailed description of the invention]

The present invention is further illustrated in conjunction with the embodiments with reference to the accompanying drawings.

Fig. 1 is the abrupt junction composition of PEH1 gene of the present invention.

Fig. 2 is the sequencer map of PEH1 gene of the present invention, and wherein a sequencer map is wild type, and b sequencer map is saltant type.

Fig. 3 is expression column diagram of the PEH1 gene of the present invention in mutant strain fro1 (t) and bright extensive 86.

[specific embodiment]

The present invention relates to a kind of rice flower organ developmental regulation gene PEH1, the gene PEH1 to have such as SEQ ID No: Nucleotide sequence shown in 1.

Addition, substitution, insertion or deletion one or more nucleotide in the SEQ ID No:1 nucleotide sequence and generate Nucleotide sequence, allele or derivative.

Addition, substitution, insertion or deletion one or more nucleotide in the gene PEH1 and its nucleotide sequence and give birth to At nucleotide sequence, allele or derivative be applied to rice development of floral organs adjust.

The invention further relates to containing a kind of above-mentioned plasmid of rice flower organ developmental regulation gene PEH1, plant expression vector, Host cell.

Plasmid, plant expression vector, the host cell containing a kind of rice flower organ developmental regulation gene PEH1 Development of floral organs applied to rice is adjusted.

The present invention further relates to a kind of protein of rice flower organ developmental regulation gene PEH1 coding, and the protein has The amino acid sequence as shown in SEQ ID No:2.

Addition, substitution, insertion or deletion one or more amino acid generates in the SEQ ID No:2 amino acid sequence Amino acid sequence or derivative.

A kind of protein of rice flower organ developmental regulation gene PEH1 coding, the protein and its amino acid The amino acid sequence or derivative that addition, substitution, insertion or deletion one or more amino acid generates in sequence are applied to rice Development of floral organs adjust.

Steps are as follows for the acquisition and its functional verification of gene of the present invention:

The PEH1 assignment of genes gene mapping: in order to which un-mixing bases are because the present invention has set up a target group first, by mutant fro1 (t) normal plant in segregating population (genotype is wild type or heterozygous) hybridizes with rice variety 9311, building positioning group Body, using chorista hybrid analysis (BSA-seq) method based on high-flux sequence, quick positional candidate gene.

The verifying of Candidate Mutant site: candidate gene is found using website Rice Genome Annotation Project Information designs a pair of specific sequencing primer in candidate locus two sides, carries out respectively to the DNA of homozygous wildtype and mutant PCR amplification is sent product to sequencing and is compared, determines candidate gene.Real-time PCR analyzes wild type and mutant in RNA On expression quantity, further determine that candidate gene.

The functional analysis of PEH1 gene: by genetic transformation, the segment comprising complete PEH1 gene is gone into mutant The test that has complementary functions is carried out in fro1 (t), obtains the transgenic paddy rice for making mutant restore normal phenotype.

It develops and dashes forward the present invention is based on a rice flower organ by unknown single recessive nuclear gene control control of early detection Variant fro1 (t) (Mao Bigang, Liu Huaqing, Chen Jianmin, Chen Jie, two rice reproductive organs mutant of Peng Yonghong, Wang Feng Morphological feature and genetic analysis Molecular Plant Breeding, 2008,6 (2): 233-238；Chen Jie, Wu Yubing, Chen Shaoyou, Liu Hua Clearly, the form generation of Wang Feng rice reproductive development mutant fro1 (t) and the Fujian Journal of Agricultural Sciench assignment of genes gene mapping, 2010,25 (6):677-683.)。

Compared with wild rice bright extensive 86, the mutant phenotype of mutant fro1 (t) is mainly manifested in: plant type is short and small, raw Growing way is weaker；Inside and outside bran piece torsional deformation obvolvent, small ear exactly likes capsicum, but small ear is closed does not bloom always, final withered It withers；Secondary grain husk is most to degenerate；Lodicule is transformed into thin leaflee；Stamen filigree expands distortion, the deformation of anther surface deflections, no flower Powder, 2-6 pieces of stamen number, part stamen are converted into that bottom is lobate, and the gynoecium shape organ of similar column cap is arranged at top；Ovary development is just Often, but most style fusions, column cap fasciation, 3-4 are a.Mutant fro1 (t) cannot be solid, can only be protected in the form of heterozygosis strain It deposits, is the mutant of single recessive gene control.

The present invention is further described below in conjunction with specific embodiment.

Embodiment 1: the acquisition of rice flower organ developmental regulation gene PEH1

Rice material: tissue culture offspring of the rice mutant fro1 (t) from rice variety bright extensive 86, positioning parent are long-grained nonglutinous rice Kind 9311.

Target group's building: choose in mutant fro1 (t) segregating population wildtype phenotype (genotype be it is wild homozygous or Heterozygosis) hybridized with rice variety 9311, construct target group F2.

The assignment of genes gene mapping: being based on F2 group, in segregant generation, carries out Phenotypic Selection, (i.e. and mutant by " mutant phenotype " Parental phenotypes are consistent) individual pick out, clip blade mixed in equal amounts extracts DNA respectively, and building obtains recessive homozygous pond；It will The individual of " wildtype phenotype " (i.e. consistent with wild-type parent phenotype) randomly selects 50 plants, and clip blade mixed in equal amounts is extracted respectively DNA constructs dominant heterozygosis pond；Single plant mutation parent's extraction DNA is chosen to be sequenced.Send parent individual and filial generation mixing pit to promise He Zhiyuan biological information Science and Technology Ltd. carries out genome sequencing, using the sequencing side IlluminaPE (paired end) Method, read length are 150bp, and sequencing depth is in 40 × left and right.It is reference with OryzasativaLcv.Nipponbare genome sequence column, is based on two pond reads Data screening variant sites.Expected gene type will be met in Liang Chizhong (is homozygous normal genotype, saltant type pond in wild type pond In be homozygous mutant genotypes) variant sites regard as Candidate Mutant site.

Screening is to 4 candidate mutant points (shown in the following table 1) altogether within the scope of full-length genome, wherein 3 between gene, Therefore it can exclude.Another Candidate Mutant site occurs to have occurred 1 in the gene coding region LOC_Os08g39420 (+800bp) The missing (TAAAAAT-TAAAAT) of a base A causes coded sequence that frameshit occurs, to cause gene protein translation Termination in advance, referring in particular to Fig. 1 (black surround indicate exon 1, left side white edge indicate 5-UTR, the right white edge indicate 3-UTR, Black arrow indicates that mutated site, No. * expression deletion segment ,-number expression a protein translation terminate).Therefore, by the LOC_ Os08g39420 gene is determined as the candidate gene of fro1 (t) mutant, is named as PEH1 (the i.e. contracting of PEPPER HULL 1 It writes).

The annotation information of 1 Candidate Mutant site of table and place gene

Embodiment 2: the determination of mutational site and target gene

In order to verify screening above as a result, designing pair of primers (5 '-in the mutational site two sides of PEH1 gene TACAGAAAGCCCTCAAGGAAGC-3 ' (as shown in SEQ ID No:3)；5 '-AACAACAAGAGGGGGAAATCAG-3'(are such as Shown in SEQ ID No:4)), PCR amplification is carried out to the genomic DNA of bright extensive 86 and mutant strain respectively, and carry out to amplified production Sequencing, referring in particular to Fig. 2, a sequencer map is wild type in Fig. 2, and b sequencer map is saltant type, and sequencing result is found in mutant strain This mutation is implicitly present in.

For the correlation for further verifying above-mentioned deletion mutation and affiliated gene is developed with rice flower organ, extract bright extensive 86 and mutant strain (each three plant) in the total serum IgEs of a little grain husk flowers of heading, then reverse transcription is at cDNA.It is special to design PEH1 gene order Different primer (5 '-CAGAAGTCAGCGGATGTAAGGT-3 ' (as shown in SEQ ID No:5)；5'- GGAATCGGCACAGCAATCAA-3'(is as shown in SEQ ID No:6)) carry out qRT-PCR related experiment.QRT-PCR reacts item Part is 95 DEG C, 30s；95 DEG C, 10s；55 DEG C, 20s；72 DEG C, 10s；40Cycles.All reactions carry out 3 biology and repeat and 3 A technology repeats, and qRT-PCR reference gene is OsActin1 (LOC_Os03g50885), and concrete outcome is shown in Fig. 3.A, B, C in Fig. 3 Respectively represent PEH1 gene in three different plants and bright extensive 86 expression difference of mutant fro1 (t), as the result is shown PEH1 transcriptional level in mutant strain is remarkably decreased compared with wild type.

Embodiment 3: complementation test verifies PEH1 gene function

According to LOC_Os08g39420 gene order, in website (https: //blast.ncbi.nlm.nih.gov/ Blast.cgi the gene upstream and downstream sequence in the OryzasativaLcv.Nipponbare BAC clone AP014964.1 comprising the gene is found on), design is complete Gene magnification primer, the primer pair separately include URS the and DRS sequence and two nucleic acid of CE Entry Vector (Vazyme) The identification sequence of restriction endonuclease a, wherein primer includes URS sequence and the identification sequence [5'-ggatc of endonuclease EcoRI TtccagagatgaattcGTTGAAGAACTCGCTGTCCGTG-3'(is as shown in SEQ ID No:7)]；Another primer include Identification sequence [the 5'-ctgccgttcgacgataagcttGTCGTCAGTCCTCGC of DRS sequence and endonuclease HindIII ATTATTCA-3'(is as shown in SEQ ID No:8)], segment 6589bp, on code area, initiation codon including whole gene Swim the regulating and controlling sequence of 1320bp promoter sequence and terminator codon downstream 419bp.

Bright extensive 86 DNA of rice variety is extracted, is usedHS DNA Polymerase with GC Buffer (TAKALA), PCR reaction condition are 94 DEG C, 5sec；98 DEG C, 10sec；60 DEG C, 30sec；72 DEG C, 7min； 32Cycles；72℃,5min.Using 1% agarose gel electrophoresis, take corresponding DNA fragments recycling.Using ClonExpress Entry One Step Cloning Kit (Vazyme) is by the segment Direct Cloning to CE Entry Vector, recombinant products Monoclonal sequencing is chosen after conversion.Will sequencing correctly clone shakes bacterium upgrading grain, with restriction endonuclease EcoRI and HindIII complete degestion, It after electrophoretic separation, extracts DNA fragmentation and is connected in pCAMBIA1300, to be built into complementing vector pCAMBIA1300+ promoter+PEH1+nos.The plasmid is transferred to Agrobacterium (Agrobacterium tumefaciens) strain by electric shock Rice transformation in LBA4404.Use being cured for the mature embryonal induction of agrobcterium-mediated transformation untransformed mutants fro1 (t) (the Soviet Army, Hu Changquan, Zhai Hongli, Yan Jingwan, Chen Jie, Wang Feng, 2003, mediated by agriculture bacillus long-grained nonglutinous rice bright extensive 86 is efficiently steady for injured tissue Determine the foundation of transformation system, Fujian Journal of Agricultural Sciench, 18 (4): 209-213).

The present invention obtains altogether independently turns 23 plants of PEH1 trans-genetic hybrid rice plant, carries out PCR identification discovery sun to transgenic plant 21 plants of plant of property, 2 plants of negative plant.Positive is turned into PEH1 trans-genetic hybrid rice plant compared with mutant of the same period, finds it Small ear shape restores normal, have complete 1 cob, 2 sterile glumes, 2 secondary grain husks, 1 lemma, 1 glumelle, 1 pair of lodicule, 6 stamens and 1 gynoecium.Male Pistil Fertility restores, solid normal.It can be seen that further being demonstrated by complementation test PEH1 gene is development of floral organs controlling gene.

In conclusion the present invention is stopped using the rice mutation of early screening, mixing pit is constructed, genome sequencing is carried out, Quickly position and determined that target gene PEH1, the gene control development of floral organs.It is interpreted and is combed by the function to PEH1 The relationship of downstream gene thereon will lay the foundation for the molecular regulation mechanism for further exploring monocotyledon flower development.

Although specific embodiments of the present invention have been described above, those familiar with the art should be managed Solution, we are merely exemplary described specific embodiment, rather than for the restriction to the scope of the present invention, it is familiar with this The technical staff in field should be covered of the invention according to modification and variation equivalent made by spirit of the invention In scope of the claimed protection.

Sequence table

<110>Fujian Province Agriculture Science Academy, Institute of Biotechnology

<120>protein and the application of a kind of rice flower organ developmental regulation gene PEH1 and its coding

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gaagcgtact catcagagaa gtcagaaagg gaggagagcc gtcgccgccg tgggatgtcg 60

ccgtcgtcgt cgtcgtcgag cacggggccg atgggggtgg cggtgtcgcc ggaggtggaa 120

gcggcgctgg cgcgcggcgg cgcggtcgtc gccctcgagt ccaccatcat ctgccacggt 180

aagctcgcgc ctcgccgcct cctccccacc ccgtctccct cccggattac atggctcact 240

ccggcgatgt gcgcccgcgg ctgggggacg cgtttggctt tcttcccatc ccaggtatgc 300

cctacccgaa gaatctccag accgccatgg aggtggaggc cgtcgtgagg gagaacgggg 360

cggttcctgc caccatagcc attctgaacg gcgtgccaca tgttggtgag tcctgtgatt 420

tttcctgtga ctcggagatc agatacatgt tcaactgttc aagtgctgga ggcgtgatcc 480

gtagttgcta gctactatga attattctcc aattaacacc ggataatttt cgtaatgtga 540

taggccttag cggcgagcaa ttgaagagct tggctgtaag tggaagacag tttcagaaga 600

cggctagaag ggatattgca catgttgtga gtaccttgca cactagaaat tggtcgttca 660

gaatgttttc caatgtgttc agttatatct gcactaaaac tttacagttg atatatgttt 720

ctcgtcttat tgcttcctgt aacgagtttt actgattgaa aggtggcctc tggtggtaat 780

ggtgcaacaa cagtttctgc cactatgttt ttcgctcata aggtcagagc caaaaacatc 840

ttactgctct tgttactgtt aatttatctt tccttgattt gatagccttt ttcttgttca 900

tctttgttac atgtagttgg tgccaatcat tcatctctat atcaaactaa agttgctctc 960

aaattcaggt tggcatacca attttcgtaa ctggagggat tggaggtgtt catagaaatg 1020

gtgaacagag taagtacctt cctccttcca atcgtgtaag gagtcccttg ctatctgcac 1080

attttacagt tgatgcttca atcagttttt ctgagttatg aatggatgct gtagccttcc 1140

attttaatat cgtttactat taccatgggt ctaaatgtgg attatagcag ctcatgcctg 1200

atagaggtca tgtggcttaa tagctctaaa agagggcaag tgaaatgaca tgtttattat 1260

atctagacga tgtaattttt tgttgattta taccatttcg atcagtttgt gtttcttcat 1320

gtgttgaaaa cttagcagaa atttaatatg cctcttgatt ttttgtaatc tagtttgtct 1380

atgcctcttg attttttgta atctagtttg ttttctttag cttttctgca tcctgttgtc 1440

attctctagt taatttaaga gtaaagtgca cgggcggtcc ttaaacttgc aggggtatgt 1500

catctaggtc tctaaactct caaaatacat atccaagtcc aagaacttgt catagtgtgt 1560

catttaggtc ccaaatcgcc ccagccccta taggatccta cgtggcactg atgtggcatg 1620

ccacacggac aagacgtggc atcattttga ctaacaaatg aaacccattt atctttttct 1680

tcttttctta tcttttcctc ctcctctttt ttttctttct tctccttttt ttttcttttc 1740

ccgagtagaa gaaaggaaaa aaaaagaaga atgagaagaa aacgaagaaa aaagaaggga 1800

aaaaggaaga aaaggacata tcacgtccat atggcatgcc atatcagcgc cacataggat 1860

tctatagggg ctatgtcgat ttgggaccta aatgatacac tttgacaagt tctggaacct 1920

agatgtgcat tttgagagtt tggggaccta gatgacacaa ccttacaagt ttaaggaccg 1980

cctatgcact ttactcttaa tttaagatgc tgatggtgtt ataccatcta ctcagtttga 2040

accctgaagc taaatattta tttatatcca ttatccacac gttctttgaa atacattacc 2100

gacagtttgt tgtatatact tgattaagac atcaattgat taagcacacg tatgtcctat 2160

taaacagttt gcactgtttg atgcagccat ggacatctcc tcagacttaa ctgaacttgg 2220

aaagactcct gtcactgtta tttcagctgg tgtgaaatct attttggaca taccacggac 2280

acttgagtac ttggtattac ctatcactcc cactcttcaa aagaatggct ctttgatctg 2340

ttaaaagtga acttccgtca gttcatgtag attgttttct tgttagaaca ttggatactt 2400

cacacactgt gtatcagtat tttctggatt ctgttcatga agtatgagtg gagcagctgt 2460

tttcaggggc tctccaacaa aattctgtag tcatatctgt tagaccactc tgatatagga 2520

atactaatgt agcattataa tttaaatatg aagtattaca aaacaaggtt tacctagtag 2580

caaagcaaaa aatgctgtgg aattggtagc ttatttttac tcctagtgat gtgctgccat 2640

aagtatgctg ttagctgcac cgttactatg ataggatgat acactgtgca aattacagag 2700

ccaggatctg atacagtaag aaaccaggaa ccagaaatta gttaccacga aaattagctc 2760

acttcctatg tattttttgt tgttatattt ccctcaagaa atagaatgct gttgcatcag 2820

aacctgaaag ttttctcatg tccatgataa ctccatccag acaagtaaaa ctgctagatc 2880

tacaactgag ggtttatagt tttccagaat catacgtttg ttcattttat tctccataat 2940

ctacgaccat ccttttcaag taagtcatga caattttctg gtgtttcttg gtatagacaa 3000

gttatttctt taggttttac tcgttaaatg tgctatattt gaaatttggt gtatttctgg 3060

ctaggcattt caatgaatgc aatgttgttt ggtgctcaat cgtataggtt aactgattgc 3120

attggcagga aactcaagga gtaacggttg ctgcttacaa aaccaatgaa tttcctgctt 3180

tctttacaga agtcagcgga tgtaaggtag aataatttgc ttatgtaaaa tgtttataag 3240

ctatatttaa tttttgtttt gtttgtttaa ggagattttc ttttctgtaa tctggtgtga 3300

atctgtataa aaaagaagag ctggaaatga gtatgggtgt tgatttttca atgttgtgta 3360

ttgtgttgat ctgaaggtcc cagaatcttt cctttatata ttttgccatg agcccttggt 3420

acagtatctt ttgtctcact gtactgcagt gttaggatgt ttgtactaac tggctcctgt 3480

ctgcttaatt taggtgccat gtcgcgttga ttctcccgaa gagtgtgcca agataatatg 3540

taagtattaa agttcgcatt taccccatct gtattcatat ctgtatcatt tgtgcgatat 3600

tggtttgaca tgcactgttg gcacatgatc cattaacccg aactgattga actccagcag 3660

acatccaata ccttttgacc tttttactct gttggttaca gaaagctact ggccaatgtt 3720

aattcagaat cacgtaaatt caacaccaat atcctattag aatctacaat tgagtcagca 3780

taagattcag aactgaaagc agcacataac acgaatcggg agttcactat tgtatagagt 3840

ttgattttat cctatgaatt actataactc aactggaaaa agaatttatt gtaacacaga 3900

gtttatcagc attgctgatt tatatctatt ttttacagat gcaaacaaga atttacatct 3960

aggatctggg attttgattg ctgtgccgat tcccaaggaa catgcagctt caggaaatgc 4020

tatagagtct gcaatacaga aagccctcaa ggaagcagag tattgtcctt actcccaacc 4080

tcctgctctc tcactttatc ttttctgtgt atttgtgata ctaacagaaa tgatatctaa 4140

tgactttcta tgttgtttca gggataaaaa tataataggc aacgcgatca ctcccttcat 4200

gcttgacaga gtgaaagtac taactggacg atcttcgcta gaagccagta tcctacaaat 4260

ttaatttcaa actttgctgt attttgaacc acgatgattt tcgtatttcc ctgtagtttt 4320

gagcattgat tagctattta tttctatttc aacatgcaat ttggcatttg gatcttttca 4380

agttggactt tatgagagaa aaatcaaaag gttcaaacgt ttagatgaat aacatctgat 4440

ttccccctct tgttgtttcc tgtgtcaata gaaacttctc cttagcatat tcagatattg 4500

cgcttgtaaa gaacaatgct cttgttggtg ctaaaattgc tgtggccctt tctgatcttc 4560

accagagagt aacaaacagt aaggttctct catcaataat aatagtaaaa tcctaattct 4620

gtaattatct ctacaactaa atacttaggc atcctggttc ataaagattt gcaaggttat 4680

ctgttcttct tgcatgtttt tagtttttac tggatttagt ctactgtctg tgtttattgt 4740

tggagctaga ccctagactt gtgatcacac tcacgtctgc tctgggccag tgatttgctt 4800

ttttatgtga gatatcacac tagcatctcc tctgcctaca tgttcaattg agacctaaca 4860

attttgaact tgctgtaaca ggatttcgga ggtctgcctt atagagtgca tgtacaaagg 4920

caagcgaggt ctgaaacatg gggcatccta ttcatcctca caatgtatgc ttgtacaact 4980

agttggaaac ttgtagcttc atccttttct ggcactatat aaatgaaacc tgaggtgcga 5040

gtgcatgatt tgtagcgaga tttgctactg tagacttaaa ttagtacatt ccactccagc 5100

ataacgtgtt cttcactttt tcgggcgtgt tcttcacttt tttgttaatc tgatcttctg 5160

ctttcacctt gaaaacttga aattaatttg aactgggcat tcagtcctcc tatttggagt 5220

gcatgaactg ttgtcaaact aatgtggaat ttgtgacttg tc 5262

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<211> 327

<212> PRT

<213>(Oryza sative)

<400> 2

Met Ser Pro Ser Ser Ser Ser Ser Ser Thr Gly Pro Met Gly Val Ala

1 5 10 15

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

20 25 30

Ala Leu Glu Ser Thr Ile Ile Cys His Gly Met Pro Tyr Pro Lys Asn

35 40 45

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

50 55 60

Val Pro Ala Thr Ile Ala Ile Leu Asn Gly Val Pro His Val Gly Leu

65 70 75 80

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

85 90 95

Lys Thr Ala Arg Arg Asp Ile Ala His Val Val Ala Ser Gly Gly Asn

100 105 110

Gly Ala Thr Thr Val Ser Ala Thr Met Phe Phe Ala His Lys Val Gly

115 120 125

Ile Pro Ile Phe Val Thr Gly Gly Ile Gly Gly Val His Arg Asn Gly

130 135 140

Glu Gln Thr Met Asp Ile Ser Ser Asp Leu Thr Glu Leu Gly Lys Thr

145 150 155 160

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

165 170 175

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

180 185 190

Lys Thr Asn Glu Phe Pro Ala Phe Phe Thr Glu Val Ser Gly Cys Lys

195 200 205

Val Pro Cys Arg Val Asp Ser Pro Glu Glu Cys Ala Lys Ile Ile Tyr

210 215 220

Ala Asn Lys Asn Leu His Leu Gly Ser Gly Ile Leu Ile Ala Val Pro

225 230 235 240

Ile Pro Lys Glu His Ala Ala Ser Gly Asn Ala Ile Glu Ser Ala Ile

245 250 255

Gln Lys Ala Leu Lys Glu Ala Glu Asp Lys Asn Ile Ile Gly Asn Ala

260 265 270

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

275 280 285

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

290 295 300

Ala Lys Ile Ala Val Ala Leu Ser Asp Leu His Gln Arg Val Thr Asn

305 310 315 320

Arg Phe Arg Arg Ser Ala Leu

325

<210> 3

<211> 22

<212> DNA

<213>(artificial sequence)

<400> 3

tacagaaagc cctcaaggaa gc 22

<210> 4

<211> 22

<212> DNA

<213>(artificial sequence)

<400> 4

aacaacaaga gggggaaatc ag 22

<210> 5

<211> 22

<212> DNA

<213>(artificial sequence)

<400> 5

cagaagtcag cggatgtaag gt 22

<210> 6

<211> 20

<212> DNA

<213>(artificial sequence)

<400> 6

ggaatcggca cagcaatcaa 20

<210> 7

<211> 43

<212> DNA

<213>(artificial sequence)

<400> 7

ggatcttcca gagatgaatt cgttgaagaa ctcgctgtcc gtg 43

<210> 8

<211> 44

<212> DNA

<213>(artificial sequence)

<400> 8

ctgccgttcg acgataagct tgtcgtcagt cctcgcatta ttca 44

Claims (8)

1. a kind of rice flower organ developmental regulation gene PEH1, it is characterised in that: the gene PEH1 has such as SEQ ID No:1 Shown in nucleotide sequence.

2. a kind of rice flower organ developmental regulation gene PEH1 according to claim 1, it is characterised in that: the SEQ ID Addition, substitution, insertion or deletion one or more nucleotide in No:1 nucleotide sequence and nucleotide sequence, the equipotential base generated Cause or derivative.

3. being carried containing a kind of plasmid of rice flower organ developmental regulation gene PEH1 of any of claims 1 or 2, plant expression Body, host cell.

4. a kind of rice flower organ developmental regulation gene PEH1 according to claim 1 or 2, it is characterised in that: the base The nucleotide sequence that is generated due to addition, substitution, insertion or deletion one or more nucleotide in PEH1 and its nucleotide sequence, The development of floral organs that allele or derivative are applied to rice is adjusted.

5. the plasmid according to claim 3 containing a kind of rice flower organ developmental regulation gene PEH1, plant expression carry The development of floral organs that body, host cell are applied to rice is adjusted.

6. a kind of protein of rice flower organ developmental regulation gene PEH1 coding, it is characterised in that: the protein has such as Amino acid sequence shown in SEQ ID No:2.

7. a kind of protein of rice flower organ developmental regulation gene PEH1 coding according to claim 6, feature exist The ammonia generated in: addition, substitution, insertion or deletion one or more amino acid in the SEQ ID No:2 amino acid sequence Base acid sequence or derivative.

8. a kind of protein of rice flower organ developmental regulation gene PEH1 coding according to claim 6 or 7, feature It is: addition, substitution, insertion or deletion one or more amino acid in the protein and its amino acid sequence and the ammonia that generates Base acid sequence or derivative, the development of floral organs applied to rice are adjusted.