Application of protein OsMPK4 in regulation and control of tillering number of rice

Abstract

The invention discloses an application of a protein OsMPK4 in regulating and controlling the tillering number of rice. The protein OsMPK4 is a protein with an amino acid sequence as shown in SEQ ID No. 1. Experiments prove that transgenic rice with increased tillering number can be obtained by introducing the gene for coding the protein OsMPK4 into Nipponbare; the gene for coding the protein OsMPK4 in a Nipponbare genome is mutated, so that transgenic rice with reduced tillering number can be obtained. Therefore, the protein OsMPK4 can regulate and control the tillering number of the rice. The method has an important application value.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of plant genetic engineering, and particularly relates to application of protein OsMPK4 in regulating number of tillers of rice. BACKGROUND

[0002] Rice (Oryza sativa L.) is one of the most important food crops, which provides the main food for about half of the world's population. Tillering is a special branch formed in the growth and development process of rice. The development process of lateral branches can be artificially divided into two development stages: one is the initiation of leaf axillary meristem and the formation of axillary bud; the other is the elongation and development of axillary bud to plant branch or dormancy. Rice tillering development is a key factor for determining the morphological development of rice, and is an important agronomic trait that directly affects the number of effective panicles per plant and thus regulates the yield of rice. In general, rice tillering is an agronomic trait that directly affects the yield of rice, and is a target trait that is focused on and improved in breeding practice. Although some important factors regulating the number of rice tillers have been identified, the mechanism of rice tiller formation is still limited. It has important theoretical significance and application value to explore more genes related to rice tillering and study the gene regulatory network involved. SUMMARY

[0003] The purpose of the present application is to increase the number of tillers of plants.

[0004] The present application first protects the application of protein OsMPK4, which can be S1) or S2):

[0005] S1) regulating the number of tillers of plants;

[0006] S2) cultivating transgenic plants with changed number of tillers;

[0007] The plant can till.

[0008] In the above application, the protein OsMPK4 can be a1), a2) or a3):

[0009] a1) the amino acid sequence is the protein shown in SEQ ID No. 1;

[0010] a2) a fusion protein obtained by connecting a tag to the N terminus or / and C terminus of the protein shown in SEQ ID No. 1;

[0011] a3) a protein related to plant tillering obtained by substitution and / or deletion and / or addition of one or several amino acid residues of the protein shown in a1) or a2).

[0012] SEQ ID No. 1 consists of 369 amino acid residues.

[0013] To facilitate the purification of the protein in a1), a tag as shown in Table 1 can be attached to the amino terminal or carboxyl terminal of the protein shown in SEQ ID No. 1.

[0014] Table 1. Sequences of tags

[0015] Tag Residue Sequence Poly-Arg 5-6 (usually 5) RRRRR FLAG 8 DYKDDDDK Strep-tag II 8 WSHPQFEK c-myc 10 EQKLISEEDL

[0016] The protein in a3) above, the substitution and / or deletion and / or addition of one or several amino acid residues is a substitution and / or deletion and / or addition of no more than 10 amino acid residues.

[0017] The protein in a3) above can be artificially synthesized, or its encoding gene can be synthesized first and then expressed biologically.

[0018] The encoding gene of the protein in a3) above can be obtained by deleting the codon of one or several amino acid residues in the DNA sequence shown in SEQ ID No. 2, and / or performing one or several base pair missense mutations, and / or connecting the encoding sequence of the tag shown in Table 1 to the 5' end and / or 3' end thereof.

[0019] The present application also protects the use of a nucleic acid molecule encoding any of the above-mentioned proteins OsMPK4, which can be S1) or S2):

[0020] S1) regulating the tiller number of a plant;

[0021] S2) cultivating a transgenic plant with changed tiller number;

[0022] The plant can tiller.

[0023] In the above use, the nucleic acid molecule encoding any of the above-mentioned proteins OsMPK4 can be a DNA molecule as follows (b1) or (b2) or (b3) or (b4):

[0024] (b1) a DNA molecule whose encoding region is as shown in SEQ ID No. 2;

[0025] (b2) a DNA molecule whose nucleotide sequence is as shown in SEQ ID No. 2;

[0026] (b3) a DNA molecule hybridizing with the DNA molecule defined in (b1) or (b2) and encoding any of the above-mentioned proteins OsMPK4;

[0027] (b4) a DNA molecule derived from rice and having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the DNA molecule defined in (b1) or (b2) and encoding any of the above-mentioned proteins OsMPK4.

[0028] The nucleic acid molecule can be DNA, such as cDNA, genomic DNA, or recombinant DNA. The nucleic acid molecule can also be RNA, such as mRNA or hnRNA, etc.

[0029] SEQ ID No. 2 consists of 1110 nucleotides, and the nucleotides shown in SEQ ID No. 2 encode the amino acid sequence shown in SEQ ID No. 1.

[0030] Those artificially modified nucleotides having 75% or more identity to the nucleotide sequence of the protein OsMPK4 isolated from the present application, as long as they encode the protein OsMPK4, are also derived from the nucleotide sequence of the present application and equivalent to the sequence of the present application.

[0031] The term "identity" as used herein refers to sequence similarity to the natural nucleic acid sequence. "Identity" includes nucleotide sequences having 75% or more, or 80% or more, or 85% or more, or 90% or more, or 95% or more identity to the nucleotide sequence of the protein OsMPK4 encoding the amino acid sequence shown in SEQ ID No. 1. Identity can be evaluated by naked eye or computer software. Using computer software, identity between two or more sequences can be expressed in percentage (%), which can be used to evaluate identity between related sequences.

[0032] In any of the above-mentioned uses, the modulating plant tiller number can be increasing plant tiller number.

[0033] In any of the above-mentioned uses, the breeding tiller number changed transgenic plant can be breeding tiller number increased transgenic plant.

[0034] In any of the above-mentioned uses, the plant can be any of the following c1) to c5): c1) dicotyledonous plant; c2) monocotyledonous plant; c3) grass plant; c4) rice; c5) rice variety Nipponbare.

[0035] The application also protects a method for cultivating a transgenic plant, which can comprise the following steps: increasing the expression amount and / or activity of any one of the above-mentioned proteins OsMPK4 in a starting plant to obtain a transgenic plant; the number of tillers of the transgenic plant is increased compared with the starting plant; and the starting plant can till.

[0036] The application also protects a plant breeding method, which can comprise the following steps: increasing the expression amount and / or activity of any one of the above-mentioned proteins OsMPK4 in a plant to increase the number of tillers of the plant.

[0037] The plant can till.

[0038] In any one of the above-mentioned methods, the "increasing the expression amount and / or activity of any one of the above-mentioned proteins OsMPK4 in a starting plant" or "increasing the expression amount and / or activity of any one of the above-mentioned proteins OsMPK4 in a plant" can be achieved by transgenesis, multiple copies, changing promoters, regulatory factors and other methods known in the art to increase the expression amount and / or activity of any one of the above-mentioned proteins OsMPK4 in a starting plant.

[0039] In the above-mentioned method, the "increasing the expression amount and / or activity of any one of the above-mentioned proteins OsMPK4 in a starting plant" or the "increasing the expression amount and / or activity of any one of the above-mentioned proteins OsMPK4 in a plant" can be achieved by introducing a nucleic acid molecule encoding any one of the above-mentioned proteins OsMPK4 into a starting plant.

[0040] In the above-mentioned method, the "introducing a nucleic acid molecule encoding any one of the above-mentioned proteins OsMPK4 into a starting plant" can be achieved by introducing a recombinant vector into the starting plant; the recombinant vector can be a recombinant plasmid obtained by inserting the nucleic acid molecule encoding any one of the above-mentioned proteins OsMPK4 into an expression vector.

[0041] The recombinant vector can be specifically the overexpression vector pMHb7Fm21GW-UBIL-OsMPK4 mentioned in the examples.

[0042] The transgenic rice can be specifically the homozygous trans-OsMPK4 gene rice UBI:OsMPK4-GFP mentioned in the examples. At this time, the starting plant can be specifically the rice variety Nipponbare.

[0043] In any one of the above-mentioned methods, the plant can be any one of the following c1) to c5): c1) a dicotyledonous plant; c2) a monocotyledonous plant; c3) a plant of the family Poaceae; c4) rice; and c5) the rice variety Nipponbare.

[0044] The experiment proves that the gene encoding the protein OsMPK4 is introduced into japonica, and the transgenic rice with increased number of tillers can be obtained. Therefore, the protein OsMPK4 can regulate the number of tillers of rice. The present application has important application value. BRIEF DESCRIPTION OF DRAWINGS

[0045] Figure 1 Subcellular localization analysis of the protein OsMPK4.

[0046] Figure 2 Target information of the OsMPK4 gene in Example 3.

[0047] Figure 3 Homozygous mutant strain OsMPK4 CRISPR -1# and OsMPK4 CRISPR -2#.

[0048] Figure 4 Plant type of homozygous trans-OsMPK4 gene rice UBI:OsMPK4-GFP, homozygous mutant strain OsMPK4 CRISPR -1# and OsMPK4 CRISPR -2#.

[0049] Figure 5 Tiller number statistics of homozygous trans-OsMPK4 gene rice UBI:OsMPK4-GFP, homozygous mutant strain OsMPK4 CRISPR -1# and OsMPK4 CRISPR -2#. DETAILED DESCRIPTION

[0050] The present application will be further described in detail below in conjunction with specific embodiments. The examples given are only for the purpose of illustrating the present application, and are not intended to limit the scope of the present application. The examples provided below can serve as a guide for further improvement by those skilled in the art, and do not in any way constitute a limitation on the present application.

[0051] In the following examples, the experimental methods are conventional methods, and are performed according to the techniques or conditions described in the literature in the art or according to the product instructions, unless otherwise specified. The materials, reagents, etc. used in the following examples can be obtained commercially, unless otherwise specified.

[0052] In the quantitative test in the following examples, three repeated experiments were set, and the average value was taken.

[0053] Example 1, discovery of protein OsMPK4 and its encoding gene

[0054] The inventors of the present application found the OsMPK4 gene from rice variety Nipponbare through a large number of experiments. The nucleotide sequence of the OsMPK4 gene in the genome of rice variety Nipponbare is shown as SEQ ID No. 2. The OsMPK4 gene encodes a protein OsMPK4, and the amino acid sequence of the protein OsMPK4 is shown as SEQ ID No. 1.

[0055] Example 2, subcellular localization analysis of the protein OsMPK4

[0056] (1) A DNA fragment with the nucleotide sequence shown as SEQ ID No. 2 was inserted into the multiple cloning site of the pBeacon vector (a kind of transient expression vector with a GFP tag at the N-terminus) to obtain a recombinant expression vector pBeacon-OsMPK4 containing the OsMPK4 gene.

[0057] (2) The recombinant expression vector pBeacon-OsMPK4 and the control vector (i.e. the pBeacon vector) were respectively transformed into protoplasts of rice variety Nipponbare by the PEG / CaCl2 transient transformation system, and then the localization information was determined by observing the GFP fluorescence signal through fluorescence confocal experiment.

[0058] The detection results are shown in Figure 1 (Bars = 5 μm, 35S: M32-GFP is the recombinant expression vector pBeacon-OsMPK4, and 35S: GFP is the control vector). The results show that the protein OsMPK4 is localized in the cytoplasm and the nucleus.

[0059] Example 3, application of the protein OsMPK4 in regulating the number of tillers of rice

[0060] I. Homozygous mutant strain OsMPK4 CRISPR -1# and OsMPK4 CRISPR -2# are obtained

[0061] Since rice is a diploid plant, when gene editing is performed, both alleles on the two homologous chromosomes in the same cell can be edited to produce the same type or different types of mutations, so the two alleles in a plant are regarded as two gene editing events. Homozygous mutant strain refers to the OsMPK4 gene on the two homologous chromosomes of the plant has the same mutation. Double allele mutant strain refers to the OsMPK4 gene on the two homologous chromosomes of the plant has mutation but different mutation forms. The number of heterozygous mutant strains refers to the OsMPK4 gene on one of the two homologous chromosomes of the plant has mutation, and the OsMPK4 gene on the other homologous chromosome has no mutation. Wild type refers to the OsMPK4 gene on the two homologous chromosomes of the plant has no mutation.

[0062] 1. Target design is performed using the OsMPK4 gene with the nucleotide sequence shown in SEQ ID No. 2, and a target is obtained. One of the targets selected in this embodiment is used for experiments (see Figure 2 ), and the target sequence is: 5'-GGATGCACTGAGGACCCTAA-3' (i.e., positions 213-232 from the 5' end of SEQ ID No. 2), the corresponding target gene is the OsMPK4 gene, and the PAM sequence corresponding to the target sequence is GGG.

[0063] 2. The DNA fragment between the restriction endonucleases AscI and SpeI of the VK005 vector is replaced with the DNA double-stranded molecule shown in positions 213-232 from the 5' end of SEQ ID No. 2, and the other positions remain unchanged, to obtain the gene editing vector VK005-OsMPK4 CRISPR .

[0064] The VK005 vector is purchased from Beijing Weishanglides Biotechnology Co., Ltd., and the item number is VK005-01.

[0065] 3. The gene editing vector VK005-OsMPK4 CRISPR is introduced into Agrobacterium EHA105 to obtain recombinant Agrobacterium.

[0066] 4. The recombinant Agrobacterium is transformed into rice variety Nipponbare to obtain the T0 generation OsMPK4 mutant.

[0067] The specific method of transforming the recombinant Agrobacterium into rice variety Nipponbare is referred to the following literature: Hiei Y, Ohta S, Komari T, Kumashiro T. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J. 1994; 6(2): 271-82. Toki S, Hara N, Ono K, Onodera H, Tagiri A, Oka S, Tanaka H. Early infection of scutellum tissue with Agrobacterium allows high-speed transformation of rice. Plant J. 2006; 47(6): 969-76.

[0068] 5. Self-pollinate the T0 generation OsMPK4 mutant to obtain T1 generation OsMPK4 mutant seeds. Germinate the T1 generation OsMPK4 mutant seeds and raise seedlings to obtain several T1 generation OsMPK4 mutants.

[0069] 6. Identification of mutation types in T1 generation OsMPK4 mutants

[0070] (1) Genomic DNA was extracted from leaves of the T1 generation OsMPK4 mutant using the improved CTAB method (Mou Z, He Y, Dai Y, et al. Deficiency in fattyacidsynthase leads to premature cell death and dramatic alterations in plant morphology. Plant Cell. 2000, 12, 405-418).

[0071] (2) After completing step (1), genomic DNA from leaves of the T1 generation OsMPK4 mutant was used as a template. PCR amplification was performed using primer pairs consisting of primers CR-OsMPK4-F: 5'-TCCATTGCCTCGTTTTTCTC-3' and primers CR-OsMPK4-R: 5'-GGCTCATTTTCACTTCTCAGGA-3', resulting in a PCR amplification product with a length of 936bp.

[0072] (3) After completing step (2), the PCR amplification products were sequenced using primers CR-OsMPK4-R. The sequencing results were compared with the gene editing target sequence of the OsMPK4 gene (i.e., positions 213-232 from the 5' end of SEQ ID No. 2) to count the mutation types.

[0073] Two homozygous mutant strains were obtained and named OsMPK4. CRISPR -1# and OsMPK4 CRISPR -2#.

[0074] OsMPK4 CRISPR The OsMPK4 gene on both homologous chromosomes of -1# has the same mutation. Specifically, the OsMPK4 gene on both homologous chromosomes has a deletion of 1 nucleotide "A" (i.e., the deletion of nucleotide 226 from the 5' end of SEQ ID No. 2), which causes a frameshift and results in the loss of function of the protein OsMPK4.

[0075] OsMPK4 CRISPRThe OsMPK4 gene of the two homologous chromosomes of #2# has the same mutation, specifically, the two homologous chromosomes have a deletion of "C" 1 nucleotide at the OsMPK4 gene (i.e. deletion of the 227th nucleotide from the 5' end of SEQ ID No. 2), thereby causing a frame shift, resulting in loss of function of the protein OsMPK4.

[0076] OsMPK4 CRISPR -1# and OsMPK4 CRISPR The mutation type of the OsMPK4 gene in #2# is shown in Figure 3 (NP is Nipponbare).

[0077] II. Obtaining homozygous OsMPK4 gene-transferred rice UBI:OsMPK4-GFP

[0078] 1. Construction of overexpression vector pMHb7Fm21GW-UBIL-OsMPK4

[0079] (1) Homologous recombination of the OsMPK4 gene shown in SEQ ID No. 2 and the pDONR221 vector (Invitrogen, item number: 12535-019) using Gateway BP clonase (Invitrogen, item number: P / N56481) to obtain the intermediate vector pDonor221-OsMPK4.

[0080] (2) After step (1) is completed, homologous recombination of the intermediate vector pDonor221-OsMPK4 and the vector pMHb7Fm21GW-UBIL (VIB-UGent Center for Plant Systems Biology, Vector ID: 3_64; the vector is a binary vector with a GFP tag at the C-terminus) using Gateway LR clonase (Invitrogen, item number: P / N56484) to obtain the overexpression vector pMHb7Fm21GW-UBIL-OsMPK4.

[0081] 2. Introduction of the overexpression vector pMHb7Fm21GW-UBIL-OsMPK4 into Agrobacterium EHA105 to obtain recombinant Agrobacterium.

[0082] 3. Transformation of the recombinant Agrobacterium into rice variety Nipponbare to obtain T0 generation OsMPK4 gene-transferred rice.

[0083] 4. Molecular detection of T0 generation OsMPK4 gene-transferred rice

[0084] (1) Genomic DNA was extracted from leaves of T0 generation rice plants that were intended to be transgenic to the OsMPK4 gene using an improved CTAB method.

[0085] (2) After completing step (1), using genomic DNA from leaves of T0 generation rice plants intended to be transgenic into the OsMPK4 gene as templates, PCR amplification was performed using primer pairs consisting of OsMPK4-OE-F: 5'-GCGACAATTACGGAACCTCC-3' and GFP-R: 5'-TGGTGCAGATGAACTTCAGGGTCAGCTTGCCGTAGGTGGCA-3' to obtain PCR amplification products. Then, the following judgment was made: if the PCR amplification product obtained from a T0 generation rice plant intended to be transgenic into the OsMPK4 gene contained a DNA fragment of 500 bp, then the T0 generation rice plant intended to be transgenic into the OsMPK4 gene was a positive seedling, i.e., a T0 generation rice plant intended to be transgenic into the OsMPK4 gene.

[0086] 5. Self-pollinate the T0 generation OsMPK4 gene-transformed rice obtained in step 4 to obtain T1 generation OsMPK4 gene-transformed rice seeds. Germinate the T1 generation OsMPK4 gene-transformed rice seeds and raise seedlings to obtain a number of T1 generation OsMPK4 gene-transformed rice plants.

[0087] 6. Molecular detection of T1 generation OsMPK4 gene transgenic rice

[0088] (1) Genomic DNA was extracted from leaves of T1 generation transgenic OsMPK4 rice using the improved CTAB method.

[0089] (2) After completing step (1), using genomic DNA from leaves of T1 generation OsMPK4 gene-transformed rice as templates, PCR amplification was performed using primer pairs consisting of OsMPK4-OE-F: 5'-GCGACAATTACGGAACCTCC-3' and GFP-R: 5'-TGGTGCAGATGAACTTCAGGGTCAGCTTGCCGTAGGTGGCA-3' to obtain PCR amplification products. Then, the following judgment was made: if the PCR amplification product obtained from a certain T1 generation OsMPK4 gene-transformed rice contained only a 500bp DNA fragment, then the T1 generation OsMPK4 gene-transformed rice was a T1 generation homozygous OsMPK4 gene-transformed rice.

[0090] The T1 generation of homozygous transgenic rice with the OsMPK4 gene was planted for several generations until the plant phenotype was stable. One of the homozygous transgenic rice plants with the OsMPK4 gene was named UBI:OsMPK4-GFP.

[0091] III. Real-time quantitative PCR detection of OsMPK4 CRISPR -1#、OsMPK4CRISPR -2# and UBI:OsMPK4-GFP

[0092] Total RNA was extracted from the tested rice plants (OsMPK4 CRISPR -1# and OsMPK4 CRISPR -2# and UBI:OsMPK4-GFP or Nipponbare) and then the relative expression of OsMPK4 gene was detected by real-time quantitative PCR (ACTIN gene as internal reference).

[0093] The results showed that, compared with Nipponbare, the relative expression of OsMPK4 gene in OsMPK4 CRISPR -1# and OsMPK4 CRISPR -2# was significantly decreased, and the relative expression of OsMPK4 gene in UBI:OsMPK4-GFP was significantly increased.

[0094] IV. Observation and analysis of the plant type of OsMPK4 CRISPR -1# and OsMPK4 CRISPR -2# and UBI:OsMPK4-GFP

[0095] The experiment was repeated three times and the average value was taken. 10 rice seedlings were observed and counted each time. The steps of each experiment were as follows: taking the seeds of the tested rice (T1 generation seeds of OsMPK4 CRISPR -1#, T1 generation seeds of OsMPK4 CRISPR -2#, UBI:OsMPK4-GFP seeds or Nipponbare seeds), washing with 50% (v / v) sodium hypochlorite solution for 20 min and washing with water for 6 times; then after soaking in water for 2 days, moving to a 37℃ incubation room for 3 days, sowing the seeds on the seedbed for seedling raising, and transplanting the rice seedlings into the paddy field at the 4-leaf stage for conventional field cultivation. At the heading stage, the plant type was observed and the tiller number was counted.

[0096] The plant type at the heading stage was observed and the tiller number was counted Figure 4 (NP is Nipponbare, Bar = 20 cm).

[0097] The plant type at the heading stage was observed and the tiller number was counted Figure 5 (NP is Nipponbare; *** indicates P < 0.001, indicating extremely significant difference).

[0098] The results showed that, compared with Nipponbare, UBI:OsMPK4-GFP showed obvious tillering phenotype and the tiller number was significantly increased, and the tiller number of OsMPK4 CRISPR -1# and OsMPK4 CRISPR -2# was significantly reduced.

[0099] Therefore, it can be seen that the protein OsMPK4 can regulate the tiller number of rice, specifically: increasing the expression amount of the protein OsMPK4 can increase the tiller number of rice; inhibiting the expression amount of the protein OsMPK4 (for example, after the OsMPK4 gene is edited, OsMPK4 gene mutation can be caused, when the OsMPK4 genes on the two homologous chromosomes are both mutated, the expression amount of the protein OsMPK4 can be reduced or even lost) can reduce the tiller number of rice.

[0100] The above has described the present application in detail. For those skilled in the art, without departing from the purpose and scope of the present application, and without unnecessary experiments, the present application can be implemented in a wide range under the same parameters, concentrations and conditions. Although the present application gives a special example, it should be understood that the present application can be further improved. In short, according to the principle of the present application, this application intends to include any change, use or improvement of the present application, including the change made by the conventional technology known in the art, which is out of the range disclosed in the present application.

Claims

1. Application of protein OsMPK4, for S1) or S2): S1) Regulates the number of plant tillers; S2) Cultivating transgenic plants with altered tiller numbers; The plant can tiller; The protein OsMPK4 is a1), a2), or a3): a1) The amino acid sequence is that of the protein shown in SEQ ID No. 1; a2) A fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of the protein shown in SEQ ID No. 1; a3) Proteins related to plant tillering obtained by substituting and / or deleting and / or adding one or more amino acid residues of the proteins shown in a1) or a2).

2. The application of the nucleic acid molecule encoding the protein OsMPK4 described in claim 1, as S1) or S2): S1) Regulates the number of plant tillers; S2) Cultivating transgenic plants with altered tiller numbers; The plant can tiller.

3. The application according to claim 2, characterized in that: The nucleic acid molecule encoding the protein OsMPK4 in claim 1 is a DNA molecule of the following type (b1) or (b2) or (b3) or (b4): (b1) DNA molecules with coding regions as shown in SEQ ID No. 2; (b2) A DNA molecule with a nucleotide sequence as shown in SEQ ID No. 2; (b3) A DNA molecule that hybridizes with the DNA molecule defined by (b1) or (b2) and encodes the protein OsMPK4 as described in claim 1; (b4) A DNA molecule derived from rice that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the DNA molecule defined in (b1) or (b2) and that encodes the protein OsMPK4 of claim 1.

4. The application according to any one of claims 1 to 3, characterized in that: The regulation of plant tiller number is to increase the number of plant tillers; The transgenic plant with altered tiller number is a transgenic plant with increased tiller number.

5. The application according to any one of claims 1 to 4, characterized in that: The plant is any one of the following c1) to c5): c1) dicotyledonous plant; c2) monocotyledonous plant; c3) grass; c4) rice; c5) rice variety Nipponbare.

6. A method for cultivating transgenic plants, comprising the following steps: increasing the expression level and / or activity of the protein OsMPK4 as described in claim 1 in a starting plant to obtain a transgenic plant; the number of tillers in the transgenic plant is increased compared to the starting plant; the starting plant is capable of tillering.

7. The method according to claim 6, characterized in that: The enhancement of the expression level and / or activity of the protein OsMPK4 of claim 1 in the starting plant is achieved by introducing a nucleic acid molecule encoding the protein OsMPK4 of claim 1 into the starting plant.

8. A plant breeding method, comprising the following steps: increasing the expression level and / or activity of the protein OsMPK4 as described in claim 1 in a plant, thereby increasing the number of tillers in the plant; The plant can tiller.

9. The method according to any one of claims 6 to 8, characterized in that: The plant is any one of the following c1) to c5): c1) dicotyledonous plant; c2) monocotyledonous plant; c3) grass; c4) rice; c5) rice variety Nipponbare.

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