CN108191962B - Medicago truncatula multi-petal control protein and coding gene and application thereof - Google Patents

Abstract

The invention discloses medicago truncatula multi-petal control protein and a coding gene and application thereof, wherein the medicago truncatula multi-petal control protein is (a1) or (a 2): (a1) a protein consisting of an amino acid sequence shown in SEQ ID No. 2; (a2) the protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO.2 and is derived from the SEQ ID NO.2 and has the same function. The gene of the invention can control the increase of the petals of medicago truncatula, the limited growth of plants, the morphological mutation of leaves and the shortening of leaf stalks.

Description

Medicago truncatula multi-petal control protein and coding gene and application thereof

Technical Field

The invention belongs to the technical field of genetic engineering, and particularly relates to medicago truncatula multi-petal control protein, and a coding gene and application thereof.

Background

Flowers are the peculiar reproductive organs of angiosperms, the normal development of the flower organs is directly related to the multiplication of species, and simultaneously, the yield, the quality, the use value and the economic benefit of flowering crops are also influenced. In addition, flower size, morphological structure, number of constituent organs, etc. are the most common morphological traits used in systematic and evolutionary phytological studies, and the study of floral organ development is also of great significance for species origin and evolution studies.

Medicago truncatula (Medicago truncatula) belongs to subfamily Papilionaceae of Leguminosae, and its flower is typical bilateral symmetric Papilionaria. In recent years, with the completion of genome sequencing of medicago truncatula and the establishment of a plurality of artificial mutant libraries, medicago truncatula has become an ideal model material for researching the development of floral organs of butterflies.

At present, some related genes of alfalfa floral organ development ABC model are cloned successively through research means of forward and reverse genetics. Researches in the butterflies Lotus japonicus (Lotus japonica) and the peas (Pisum sativum) also find that the butterflies and the dicotyledonous model plant Arabidopsis thaliana have certain conservation in the floral organ development regulation mechanism. However, the development regulation of the butterfly flower organ is more complicated due to the specificity of the butterfly flower organ in morphology and development, the problems of gene doubling, gene function differentiation, gene expression mode change, protein function domain deletion or acquisition and the like of the butterfly flower subfamily plant.

Therefore, there is a need to provide a plant multi-petal control gene and a protein encoded by the same.

Disclosure of Invention

In view of the above, the invention provides medicago truncatula multi-petal control protein, and a coding gene and application thereof.

In order to solve the technical problem, the invention discloses medicago truncatula multi-petal control protein which is (a1) or (a 2):

(a1) a protein consisting of an amino acid sequence shown in SEQ ID No. 2;

(a2) the protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO.2 and is derived from the SEQ ID NO.2 and has the same function.

The invention also discloses a gene for coding the protein.

Optionally, the gene is a DNA molecule as described in any one of (b1) to (b3) below:

(b1) the coding region is a DNA molecule shown as SEQ ID NO. 1;

(b2) a DNA molecule shown as SEQ ID NO. 3;

(b3) a DNA molecule which hybridizes under stringent conditions to the DNA sequence defined in (b1) or (b2) and which encodes the protein of claim 1;

(b4) a DNA molecule derived from medicago truncatula and having more than 90% homology with the DNA sequence defined in (b1) or (b2) or (b3) and encoding the protein of claim 1.

The invention also discloses a recombinant expression vector, an expression cassette, a transgenic cell line or a recombinant bacterium containing the gene.

The invention also discloses application of the protein or the gene in regulation and control of a multi-petal phenotype of a plant.

Optionally, the plant is a dicot.

Optionally, the dicot is a legume.

Optionally, the leguminous plant is medicago truncatula.

The invention also discloses a method for increasing the number of plant petals, which comprises the following steps: inhibiting the activity of the protein and/or the expression level of the gene in the target plant to obtain a transgenic plant with increased petals.

The invention also discloses application of the protein, the gene or the method in plant breeding.

Compared with the prior art, the invention can obtain the following technical effects:

1) the invention clones a gene for controlling the increase of the petals of the medicago truncatula, the limited growth of the plant, the morphological mutation of the leaves and the shortening of the petiole in the medicago truncatula for the first time, and the gene provides a gene sequence for cloning related genes by utilizing homologous genes for other species for the first time.

2) The cloned gene can also provide evidence for the gene research of petal increase, leaf form mutation and petiole shortening of leguminous plants such as medicago truncatula and other dicotyledonous plants.

Of course, it is not necessary for any one product in which the invention is practiced to achieve all of the above-described technical effects simultaneously.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention and not to limit the invention. In the drawings:

FIG. 1 is a floral phenotype plot of alfalfa R108 wild-type flowers and alfalfa pis multi-petal mutants of the present invention; wherein, a is a side view of a wild type bud at the bud stage, B and C are a side view and a top view, respectively, of a wild type young flower, D and E are a side view and a top view, respectively, of a wild type mature flower, F is a side view of pis mutant bud, G and H are a side view and a top view, respectively, of a pis mutant young flower, I and J are a side view and a top view, respectively, of a pis mutant mature flower (the J-diagram arrows indicate that inside the mutant mature flower, there is calyx growth in addition to petal increase), and K is a late plant growth stage; l is a wild type compound leaf petiole, M is a mutant compound leaf petiole, N is a wild type leaf, O is a mutant leaf, P is a leaf petiole at the top of the wild type compound leaf, and Q is a leaf petiole at the top of the mutant compound leaf;

FIG. 2 is a diagram showing the structure of the PIS gene of the present invention and the detection of the PIS gene expression level in the mutant; wherein, A represents a PIS gene structure diagram (comprising four exons and three introns) and a Tnt1 insertion position in each PIS mutant, and B represents a DNA level identification PIS mutant; c represents RT-PCR detection for PIS full-length gene expression level detection in each mutant.

Detailed Description

The following embodiments are described in detail with reference to the accompanying drawings, so that how to implement the technical features of the present invention to solve the technical problems and achieve the technical effects can be fully understood and implemented.

Medicago truncatula R108 (wild type): the Nobel Foundation.

pis mutant: the Nobel Foundation. pis-1(NF10617) pis-2(NF 21657);

pis-3(NF16829)；

example 1 acquisition of 1pis mutants and phenotypic analysis

A mutant NF10617 with Increased Petals and complete deletion of stamens and pistils is obtained by screening the Medicago truncatula Tnt1 insertion mutant library and is named as Petals incorporated Severely (pis).

A detailed phenotypic observation of the pis mutant is shown in FIG. 1. FIG. 1A is a side view of a wild type bud at the time of the small bud; b and C are respectively a side view and a top view of the wild type young flower; d and E are respectively a side view and a top view of a wild-type mature flower; f is pis mutant small bud side view; g and H are respectively a side view and a top view of the pis mutant young flower; i and J are respectively a side view and a top view of the pis mutant mature flower (the red arrow in the J-panel indicates that inside the mutant mature flower there is also calyx-to-grow in addition to the increase in petals). Mutant pis exhibited an increased bud phenotype throughout the flower development period compared to wild type. K-Picture shows that in late floral development, the pis mutant lost the ability to grow indefinitely compared to the wild type, and the mutation was limited growth by a terminal flower that terminated with floral organs.

In addition, the leaves of the mutant were also altered, and the leaf stalks of the multiple leaves were significantly shorter than those of the wild type (FIG. 1L, M), the leaf edges were depressed (FIG. 1L-Q), and the leaf stalks at the top of the multiple leaves were almost absent (FIG. 1P, Q).

EXAMPLE 2 cloning of PIS Gene

According to Tnt1 website (NF10617 published by https:// media-mutatedDesigning corresponding primers for the flanking sequences of (1),

NF10617-F:5’-CCTCCTCTAACCTGCTCCA-3’；

NF10617-R:5’-TCACCACCTCTTTCCCATTCA-3’；

and (3) detecting the linkage of the mutant and the gene. Finally, we found that the mutant linked gene PIS, detected at DNA level, amplified a band about 5.5K longer than the wild type at the insertion site compared to the wild type (FIG. 2B), and determined as Tnt1 sequence by sequencing alignment. The NF10617 mutant has Tnt1 inserted in the first exon of the gene. To demonstrate that the PIS gene is the control gene for the mutant, we ordered two additional mutants of this gene: NF21657(pis-2 inserted on the first exon), NF16829(pis-3 inserted on the fourth exon) and the previous NF10617 were named pis-1 (see FIG. 2A). The phenotype of the homozygous mutants of both material pis-2 and pis-3 was observed and found to be identical to the previous pis-1 phenotype. In PIS-1PIS-2PIS-3 homozygous mutant, PIS gene full-length expression was completely deleted (FIG. 2C). The PIS gene is proved to be a control gene of a multi-petal phenotype.

Example 3 acquisition of PIS Gene and protein PIS encoded thereby

1. RNA from alfalfa R108 (wild-type) flourishing flowers was extracted and reverse transcribed into cDNA.

2. And (3) taking the cDNA obtained in the step (1) as a template, and amplifying by adopting a primer pair consisting of a primer PIS-F and a primer PIS-R to obtain a PCR amplification product.

PIS-F：5’-CACCATGGTTGGTGACAAAGGAGA-3’；

PIS-R：5’-TCACATCAAACCACCACCAC-3’。

3. Sequencing the PCR amplification product obtained in the step 2 to obtain a coding region sequence of the target gene, wherein the coding region sequence is represented by a sequence 1 in a sequence table, and the protein is represented by a sequence 2 in the coding sequence table. Consists of 1009 amino acid residues. The CDS of the gene is compared at an alfalfa website (http:// blast.jcvi.org/Medicago-Blast /), so as to obtain the genome sequence of the PIS gene, as shown in a sequence 3. The protein shown in the sequence 2 of the sequence table is named as PIS, and the coding gene of the PIS is named as PIS gene.

The invention finds a Tnt1 insertion mutant with increased petals from a Medicago truncatula Tnt1 insertion mutant library, and clones a control gene PIS with the increased petals through flanking sequence analysis. To demonstrate that PIS is a multi-petal control gene, we screened two additional insertional mutants of the PIS gene back in the mutant pool and found that these two mutants exhibited the same phenotype as the previous mutants. The invention discloses a theoretical basis for researching the increase of petals of the PIS gene in the plant.

While the foregoing description shows and describes several preferred embodiments of the invention, it is to be understood, as noted above, that the invention is not limited to the forms disclosed herein, but is not to be construed as excluding other embodiments and is capable of use in various other combinations, modifications, and environments and is capable of changes within the scope of the inventive concept as expressed herein, commensurate with the above teachings, or the skill or knowledge of the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.

Sequence listing

<110> institute of biotechnology of Chinese academy of agricultural sciences

<120> medicago truncatula multi-petal control protein, and coding gene and application thereof

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catcatttac aagagccacc accacaaaat ttacgttttc cacttaggaa agaagtgttt 1680

gatgatttac cacaaggtaa agaattcttc ttcacaacaa cttctgaacc attggattgt 1740

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aatagggtcg tttcaagatt ctgccacgat gaaatggtaa ttactagaaa gcccttttca 1920

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tatatccttg gctactatgc aataggaaat aaggtaacat tttgtgcact tagtaaatca 2160

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gaaagattca aagccttagg accatgtttt agaattggta ttttattatc aatgttaagc 2280

aaagcttgtt ccaacatgca aaagggtagt tttgtttata gtgatttcga aaggtatagt 2340

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ataagagcaa aacccttgaa cattgaacaa cttgtagaag ccctaaagta tgtaacaaaa 2580

gctttggttg cattacatga cttatcattc atgcatagag atctatgttg ggaaaaggtt 2640

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ggaagacacg cgccagaaat ggagcgtgga ttacatggtg tgaaagtaga tgtatggagt 2820

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cagaattggt gtatggaaca gaatccagaa caaagaccaa ctgcagcgga ttgttatcat 2940

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Gln Gln Phe Ser Ser Ser Ser Ser Pro Lys Pro Gln His Glu Glu Ser

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Phe Ile Ser Thr Pro Leu Tyr Val Ser Ser Gly Gly Ala Ser Ser Ser

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Pro Leu Ala Ala Ser Ser Ser Glu Thr Thr Ser Ser Gln Ser His Asp

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His Ser Pro Met Pro Cys Gln Glu Gly Asn Arg Leu Gln Glu Leu Asp

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Trp Lys Pro Asn Glu Met Leu Trp Leu Ala Arg Ala Trp Lys Glu Gln

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Tyr Gln Thr Ser Ser Glu Thr Ser Ser Arg Thr Glu Gln Glu Met Gly

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Met Ser Arg Gly Lys Thr Arg Ala Asp Lys Asp Lys Glu Val Ala Glu

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Phe Leu Asn Lys His Gly Val Asn Arg Asp Ala Lys Thr Ala Gly Thr

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Lys Trp Asp Asn Met Leu Gly Glu Phe Arg Lys Val Tyr Glu Trp Glu

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

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Pro Tyr Glu Arg Lys Leu His Arg Leu Pro Ala Ser Phe Asp Glu Glu

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Val Phe Glu Glu Leu Ser Gln Phe Met Gly Ser Arg Met Arg Ser Ser

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Asp Asp Asp Leu Pro Leu Ser Ala Arg Thr Lys Gln Leu Ala Met Thr

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Ser Gly Gly Gly Glu Pro Phe Phe His Gly His Arg Gly Asn Ile Leu

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

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Glu Leu Arg Arg Ile Gly Lys Ile Arg Met Val Trp Glu Glu Ser Val

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Gln Asn Ser Ser Phe Leu His Ala Asp Glu Leu Thr Cys Phe Asp Asp

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

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Gly Phe Ser Val Asp Arg Phe Val Ser Gly Gln Gln Val Lys Val Phe

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Gly Arg Arg Lys Ser Ser Ser Thr Ser Ser Gly Phe Ala Glu Arg Val

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Phe Arg Asp Pro Thr Glu Tyr Tyr Met Asp Cys Leu Leu Arg Ile Ser

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Ser Pro Gln Ser Leu Pro Thr Leu Phe Glu Leu Lys His His Leu Gln

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Asp Asp Leu Pro Gln Gly Lys Glu Phe Phe Phe Thr Thr Thr Ser Glu

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Pro Leu Asp Cys Arg Ser Ile Ile Tyr Asp Ile Val Gly Pro Ile Ile

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Arg Thr Asn Asn Asn Ile Asn Asn Pro Ser Thr Leu Pro Phe Ser Ser

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Arg Asp Ser Phe Ile Gly Leu Trp Asp Asp Cys Ile Asn Arg Val Val

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Ser Arg Phe Cys His Asp Glu Met Val Ile Thr Arg Lys Pro Phe Ser

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Ser Phe Leu Ser Gln Ser Gln Asp Thr Leu Leu Leu Gln Asp Glu Trp

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Pro Asn Val Ser Gly Phe Val Asn Asn Phe Cys Leu Trp Arg Gly Glu

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Glu Cys Glu Glu Phe Lys Glu Asn Ile Gln Asn Pro Ser Ser Thr Ile

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Met Gln Lys Leu Leu Trp Ser Tyr Leu Asp Leu Pro Tyr Ile Leu Gly

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Tyr Tyr Ala Ile Gly Asn Lys Val Thr Phe Cys Ala Leu Ser Lys Ser

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Gln Glu Asp Gly Lys Ile Ile Arg Thr Asp Leu His Gln Val Asn Leu

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Thr Thr Pro Ser Glu Arg Phe Lys Ala Leu Gly Pro Cys Phe Arg Ile

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Gly Ile Leu Leu Ser Met Leu Ser Lys Ala Cys Ser Asn Met Gln Lys

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Gly Ser Phe Val Tyr Ser Asp Phe Glu Arg Tyr Ser Phe Gly His Gly

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Val Ile Ile Glu Met Thr Pro Asn Thr Cys Lys Arg Val Phe Leu Glu

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Lys Arg Lys Phe Ser Ser Val Lys Glu Val Tyr Glu Ile Leu Asp His

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Arg Ile Pro His Ser Glu Tyr Leu Val Lys Val Leu Glu Asn Asn Met

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Ser Leu Val Phe Lys Pro Arg Gly Ile Arg Ala Lys Pro Leu Asn Ile

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Glu Gln Leu Val Glu Ala Leu Lys Tyr Val Thr Lys Ala Leu Val Ala

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Leu His Asp Leu Ser Phe Met His Arg Asp Leu Cys Trp Glu Lys Val

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Thr Met Arg Asn Asp Arg Glu Gly Gly Glu Trp Phe Val Ser Gly Phe

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Asp Glu Ala Ala Gly Ala Pro Glu Leu Gly Lys Tyr Val Lys Glu Asn

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His Glu Gly Gly Met Val Glu Arg Gly Arg His Ala Pro Glu Met Glu

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Arg Gly Leu His Gly Val Lys Val Asp Val Trp Ser Val Gly Tyr Leu

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Ile Met Thr Cys Gly Leu Val Asn Val Pro Lys Met Leu Arg Glu Leu

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Gln Asn Trp Cys Met Glu Gln Asn Pro Glu Gln Arg Pro Thr Ala Ala

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Asp Cys Tyr His His Leu Leu Gln Leu Gln Ser Thr Leu Leu Val Ser

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Gly Gly Val Val Ala Gly Ser Cys Gly Val Ser Gly Gly Gly Gly Leu

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caacaactaa gaaaagggaa atatgtaagt cctgtttgga aacctaatga aatgctatgg 600

ttagcaagag catggaaaga acaataccaa acaagctcag aaacatcttc aagaactgaa 660

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ggtaaaagct attttagact ttcaccttat gaaagaaagt tgcataggtt accagcttct 900

tttgatgaag aagtttttga agaactttca cagtttatgg gatcaagaat gagatcttct 960

catggtggtg gtggtggtgg tagagttggt tcttcttttg tttcttgtga tgaagctaga 1020

acaagatctc ttcctcctcc tagacctttc aaagatgatg atcttcctct ttcaggttta 1080

ttttatatga aaaaaattta atttatgaac tttttttcct cacatttagt taaaaccctt 1140

tttttatttt tcaccaaatc ataatcttca atacactagc caaaatttga aacagtaaag 1200

ttttgtcttt tatatatatt tcttagattc ttagatatat tggacaagct attattatta 1260

ttactactac tattattatt attattatta ttattattat tattattatt attattatta 1320

ttattattat tactcttcac atttaattaa actccctttt tctattaaat taaaattcct 1380

catccattat aaatttgtaa cctactcata atctttaatg cacttgtgaa attaattgaa 1440

acagttaagt ttttatttta ttttaacttc ttatagaata aacagtagtt tgagacattc 1500

atgaaagttt tggaaggcaa gtatattatt tactgtttct gttattctct ctgactgttt 1560

caactaaaaa tactgtttga cattgtttca gggaaagaat gttttagaaa aggtggaaga 1620

aaaaaaaaga ttagtccaaa aatattaaaa ataaataaat aaataacaat taaccatttt 1680

ttttagtaac caaaaaaaaa ttaaccattt ttccaatcca aaataaatca attttttttt 1740

caactaatat tgtgtcatta ttacagctag gacaaaacaa ttggcaatga caagtggtgg 1800

aggtgaacca ttttttcatg gtcatagagg aaacatatta gggttagatt cactaatgga 1860

aatttcaaca ccttcttcta gctctaaaga gctaagaaga attggtaaga taagaatggt 1920

atgggaagaa tcagtgagtt tatggggtga agaaggtgaa gttcatagag ggagaataag 1980

ggttcaaaat tcaagctttt tacatgctga tgaactcact tgttttgatg atgcaatggt 2040

gatttgtcct ttggaatctt ttgaagatgg tcctttcaaa ggtttttcag ttgatagatt 2100

tgtttctgga caacaagtta aagtttttgg tagaagaaaa tcttcttcaa cttctagtgg 2160

tattaaacta aaactatcaa taatttttct ttggtttgtt tttacatgtt ttaggattta 2220

attttaatta aattttattc atccttatat ttttcaggtt ttgctgaaag agttcaacct 2280

atcaacaaac caattcccat aagatgtgag ttccttaatt tatgtagcta gcattattaa 2340

gactcattct ttaatcaata gagtttatga gtaacacata tattaaagtt tgcaatcata 2400

tcatttttaa ttgaataaca tattttttta atttcatttc attcaaacag aatatataag 2460

tattttttga acaatattgc aaacttgtag catttcattc attcattgcc aataatttgc 2520

tcaacaaata ggatgaagaa gttaaggaat agttgattca taatgatgtt ttgttaagag 2580

tttgaactcc actcctacat ttttcttttt aaaatttgtt caaaaatctt ggttctaaat 2640

taagggttac attaatttga aggctttaca cttgcaattt gcattcctat ttttgcaatt 2700

gcatatagca atcacaattt aaaaccatgt caaactaaaa ctaaaatcat gtacaattgc 2760

tttttactta taattgtata tagtttattt ttacataaag caactacaat tgcaactaca 2820

ttatcttcat ttctatgcat atttatctca atttagagtt atgataaatt atggattaaa 2880

aaaaaagaat tataataaat taaagacatg gatatgcttt ttactttaca ctgaagcatt 2940

aaattatcca gcaagtatta tagtgcttct aattaaattt ggcagaatca taattaattt 3000

atggttggtg tatttaattt gcagcaattg ttccattgga ttttagagac ccaactgaat 3060

actacatgga ttgtctcctc cgtatttcat caccacaatc acttccaaca ttatttgaat 3120

taaagcatca tttacaagag ccaccaccac aaaatttacg ttttccactt aggaaagaag 3180

tgtttgatga tttaccacaa ggtaaagaat tcttcttcac aacaacttct gaaccattgg 3240

attgtagatc aataatctat gacattgtgg gccccattat tagaaccaat aacaacatta 3300

acaaccctag tactctccca ttttctagta gagactcttt cattggtcta tgggatgatt 3360

gcatcaatag ggtcgtttca agattctgcc acgatgaaat ggtaattact agaaagccct 3420

ttttatcatt ttcatcacaa tcacaagata cattgttgtt gcaagatgaa tggcctaatg 3480

tgagtggttt tgtgaacaat ttttgtttat ggagaggtga agaatgtgaa gaatttaaag 3540

agaatattca aaacccatca tcaactatca ttcaaaaatt actatggagc tatttggatt 3600

taccttatat ccttggttac tatgcaattg gaaataaggt aacattttgt gcacttagta 3660

aatcacaaga agatgggaaa atcattagaa ctgatttaca tcaagtgaat ctaacaacac 3720

caagtgaaag attcaaagcc ttaggaccat gttttagaat cggcattttg ttatctatgt 3780

taagcaaagc ttgttccaac atgcaaaagg gtagttttgt ctatagtgat ttcgaaaggt 3840

atagttttgg tcaaggtgtt ttaatcgaaa tgacaccaaa cacatgcaaa agggtatttt 3900

tggaaaagag aaaattctca tcagtgaaag aagtttacga gatattagat catagaatac 3960

cacatagtga gtatttagtc aaagttcttg aaaacaacat gagtttggta tttaaaccta 4020

gaggaataag agcaaaaccc ttaaacattg aacaacttgt agaagcccta aaatatgtaa 4080

caaaagcttt agttgcatta catgacttat cattcatgca tagagatcta tgttgggaaa 4140

aggttacaat gagaaatgat agagaaggag gagaatggtt tgtgagtggt ttcgacgaag 4200

cggccggagc gccggagcta gggaaatatg taaaggataa tcatgaaggt ggtatggtgg 4260

agcgtggaag acacgcgcca gaaatggagc gtggattgca tggtgtgaaa gttgatgtgt 4320

ggagtgttgg gtatttgata atgacatgtg ggttggtgaa tgtgccaaag atgttaagag 4380

agttacagaa ttggtgtatg gaacagaatc cagaacaaag accaactgca gcggattgtt 4440

atcatcactt gttacaactt cagtcgactt tgttggtgtc aggtggtgtt gttgctggtt 4500

cttgtggtgt ttctggtggt ggtggtttga tgtga 4535

<210> 4

<211> 19

<212> DNA

<213> Artificial sequence (Artificial sequence)

<400> 4

cctcctctaa cctgctcca 19

<210> 5

<211> 21

<212> DNA

<213> Artificial sequence (Artificial sequence)

<400> 5

tcaccacctc tttcccattc a 21

<210> 6

<211> 24

<212> DNA

<213> Artificial sequence (Artificial sequence)

<400> 6

caccatggtt ggtgacaaag gaga 24

<210> 7

<211> 20

<212> DNA

<213> Artificial sequence (Artificial sequence)

<400> 7

tcacatcaaa ccaccaccac 20

Claims (3)

1. The application of a protein or a coding gene thereof in regulating and controlling the multi-petal phenotype of a plant is characterized in that the protein is a protein with an amino acid sequence shown as SEQ ID NO. 2; the regulation is to inhibit or inactivate the expression of the protein coding gene in the plant, so that the petals of the plant are increased; the plant is medicago truncatula.

2. The use according to claim 1, wherein the coding gene is a DNA molecule as described in any one of (b1) to (b 4):

(b1) the coding region is a DNA molecule shown as SEQ ID NO. 1;

(b2) a DNA molecule shown as SEQ ID NO. 3;

(b3) a DNA molecule which hybridizes under stringent conditions to the DNA sequence defined in (b1) or (b2) and which encodes the protein of claim 1;

(b4) a DNA molecule derived from medicago truncatula and having more than 90% homology with the DNA sequence defined in (b1) or (b2) or (b3) and encoding the protein of claim 1.

3. A method for increasing plant petals, which is characterized by comprising the following steps: inhibiting the activity and/or gene expression level of the protein as claimed in claim 1 in a target plant to obtain a transgenic plant with increased petals, wherein the plant is medicago truncatula.

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