Application of MsPIF8 protein and coding gene thereof in regulating and controlling growth of medicago sativa under short day

Abstract

The invention discloses an application of MsPIF8 protein and a coding gene thereof in regulating and controlling the growth of medicago sativa under short day. The MsPIF8 protein is any one of the following proteins: a) a protein with an amino acid sequence as shown in SEQ ID NO.2; b) a fusion protein obtained by connecting a label to the N terminal and / or C terminal of the protein as shown in SEQ ID NO.2; c) a protein which is obtained by substitution and / or deletion and / or addition of one or more amino acid residues of the amino acid sequence as shown in SEQ ID NO.2 and has the same function; and d) a protein which has 75% or more than 75% of identity with the amino acid sequence as shown in SEQ ID NO.2 and has the same function with the amino acid sequence as shown in SEQ ID NO.2. Experiments prove that overexpression of the MsPIF8 gene in medicago sativa can improve the plant height and the number of stem nodes of medicago sativa under a short-day condition. The invention has important significance for improving the yield of medicago sativa.

Description

Technical Field

[0001] The present invention relates to the field of plant genetic engineering, and in particular to application of MsPIF8 protein and its encoding gene in regulating the growth of alfalfa under short-day conditions. Background Art

[0002] Alfalfa (Medicago sativa L.) is an excellent perennial legume forage. Known as the "King of Forages," it boasts high crude protein content, rich nutritional value, and excellent palatability. It plays a vital role in agricultural and animal husbandry production. As living standards continue to improve, demand for livestock products is also increasing. Cultivating high-quality alfalfa and continuously increasing alfalfa yields are of practical significance for promoting the development of the livestock industry.

[0003] Light provides energy for plants; it also serves as an important environmental signal, regulating plant growth and development, such as photomorphogenesis, dark morphogenesis, and shade response. Changes in light quality and photoperiod can affect plant growth. Alfalfa is a long-day plant. When the photoperiod shifts in autumn, it experiences fall dormancy, altering its growth and morphology. This affects its cold tolerance and wintering ability, ultimately reducing yield and quality, negatively impacting the development of animal husbandry. Summary of the Invention

[0004] The purpose of the present invention is to provide MsPIF8 protein and its encoding gene and application.

[0005] In a first aspect, the present invention claims a protein.

[0006] The protein claimed in the present invention is derived from alfalfa and is named MsPIF8. The MsPIF8 protein is the protein shown in a) or b) or c) or d) below:

[0007] a) the amino acid sequence is the protein shown in SEQ ID NO. 2;

[0008] b) a fusion protein obtained by connecting a tag to the N-terminus and / or C-terminus of the protein shown in SEQ ID NO. 2;

[0009] c) a protein having the same function 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;

[0010] d) A protein having 75% or more identity with the amino acid sequence shown in SEQ ID NO. 2 and having the same function.

[0011] In the protein described in b) above, the tag refers to a polypeptide or protein that is fused and expressed with the target protein using in vitro DNA recombination technology to facilitate the expression, detection, tracing, and / or purification of the target protein. The tag can be a Flag tag, His tag, MBP tag, HA tag, myc tag, GST tag, and / or SUMO tag, etc.

[0012] In the protein described in c) above, the substitution and / or deletion and / or addition of one or several amino acid residues is substitution and / or deletion and / or addition of no more than 10 amino acid residues, or substitution and / or deletion and / or addition of no more than 9 amino acid residues, or substitution and / or deletion and / or addition of no more than 8 amino acid residues, or substitution and / or deletion and / or addition of no more than 7 amino acid residues, or substitution and / or deletion and / or addition of no more than 6 amino acid residues, or substitution and / or deletion and / or addition of no more than 5 amino acid residues, or substitution and / or deletion and / or addition of no more than 4 amino acid residues, or substitution and / or deletion and / or addition of no more than 3 amino acid residues, or substitution and / or deletion and / or addition of no more than 2 amino acid residues, or substitution and / or deletion and / or addition of no more than 1 amino acid residue.

[0013] In the protein described in d) above, the identity refers to amino acid sequence identity. Amino acid sequence identity can be determined using a homology search site on the Internet, such as the BLAST page on the NCBI homepage. For example, in Advanced BLAST 2.1, a search can be performed using blastp as the program, setting the Expect value to 10, all filters to OFF, BLOSUM62 as the matrix, and setting the Gap existence cost, Per residue gap cost, and Lambda ratio to 11, 1, and 0.85 (default values), respectively. The identity value (%) can then be obtained. The identity includes amino acid sequences having 75% or greater, 80% or greater, 85% or greater, 90% or greater, or 91% or greater, or 92% or greater, or 93% or greater, or 94% or greater, or 95% or greater, or 96% or greater, or 97% or greater, or 98% or greater, or 99% or greater homology to the amino acid sequence of SEQ ID NO. 2 of the present invention.

[0014] The proteins described in a) or b) or c) or d) above can be synthesized artificially, or their encoding genes can be synthesized first and then expressed biologically.

[0015] In a second aspect, the present invention claims biological materials related to the above-mentioned MsPIF8 protein.

[0016] The biological material related to the MsPIF8 protein claimed in the present invention is any one of the following A1) to A8):

[0017] A1) a nucleic acid molecule encoding the MsPIF8 protein;

[0018] A2) an expression cassette containing the nucleic acid molecule described in A1);

[0019] A3) a recombinant vector containing the nucleic acid molecule described in A1);

[0020] A4) a recombinant vector containing the expression cassette described in A2);

[0021] A5) a recombinant microorganism containing the nucleic acid molecule described in A1);

[0022] A6) a recombinant microorganism containing the expression cassette described in A2);

[0023] A7) a recombinant microorganism containing the recombinant vector described in A3);

[0024] A8) A recombinant microorganism containing the recombinant vector described in A4).

[0025] Furthermore, the nucleic acid molecule in A1) is the gene shown in 1) or 2) below:

[0026] 1) The coding sequence is the DNA molecule shown in SEQ ID NO.1;

[0027] 2) A DNA molecule that has 75% or more identity with the nucleotide sequence defined in 1) and encodes the MsPIF8 protein.

[0028] Those skilled in the art can readily mutate the nucleotide sequence encoding the MsPIF8 protein of the present invention using known methods, such as directed evolution and point mutagenesis. Artificially modified nucleotide sequences that share 75% or greater identity with the isolated MsPIF8 nucleotide sequence of the present invention are derived from and are equivalent to the nucleotide sequence of the present invention, as long as they encode the MsPIF8 protein and have the same function.

[0029] As used herein, the term "identity" refers to sequence similarity to a naturally occurring nucleic acid sequence. "Identity" includes nucleotide sequences that are 75% or greater, or 80% or greater, or 85% or greater, or 90% or greater, or 95% or greater identical to the nucleotide sequence of the present invention encoding the protein consisting of the amino acid sequence set forth in SEQ ID NO. 2. Identity can be assessed visually or using computer software. Using computer software, the identity between two or more sequences can be expressed as a percentage (%), which can be used to assess the identity between related sequences.

[0030] The expression cassette is a DNA capable of expressing the MsPIF8 protein in a host cell. The DNA may include not only a promoter for initiating MsPIF8 transcription but also a terminator for terminating MsPIF8 transcription. Furthermore, the expression cassette may also include an enhancer sequence. Promoters that can be used in the present invention include, but are not limited to, constitutive promoters, tissue-, organ-, and development-specific promoters, and inducible promoters. Suitable transcription terminators include, but are not limited to, the Agrobacterium nopaline synthase terminator (NOS terminator), the cauliflower mosaic virus CaMV 35S terminator, the tml terminator, the pea rbcS E9 terminator, and the nopaline and octopine synthase terminators.

[0031] The vector can be a plasmid, cosmid, phage, or viral vector. The recombinant vector can be a vector containing the MsPIF8 gene expression cassette constructed using an existing plant expression vector. The plant expression vector includes binary Agrobacterium vectors and vectors that can be used for plant microprojectile bombardment, such as pAHC25, pBin438, pCAMBIA1302, pCAMBIA2301, pCAMBIA1301, pCAMBIA1300, pBI121, pCAMBIA1391-Xa, or pCAMBIA1391-Xb. The plant expression vector can also contain the 3′ untranslated region of the exogenous gene, i.e., the polyadenylation signal and any other DNA fragments involved in mRNA processing or gene expression. The polyadenylation signal can guide the addition of polyadenylation to the 3' end of the mRNA precursor, such as the non-translated region transcribed at the 3' end of Agrobacterium crown gall induction (Ti) plasmid genes (such as the nopaline synthase gene Nos) and plant genes (such as soybean storage protein genes) all have similar functions. When using the gene construction plant expression vector of the present invention, enhancers can also be used, including translation enhancers or transcription enhancers. These enhancer regions can be ATG start codons or adjacent region start codons, etc., but must be identical to the reading frame of the coding sequence to ensure the correct translation of the entire sequence. The sources of the translation control signals and start codons are wide, and can be natural or synthetic. The translation initiation region can come from a transcription initiation region or a structural gene. To facilitate the identification and screening of transgenic plant cells or plants, the plant expression vectors used can be modified to include genes encoding enzymes or luminescent compounds that can be expressed in plants (such as the GUS gene, luciferase gene), antibiotic marker genes (such as the nptII gene, which confers resistance to kanamycin and related antibiotics; the bar gene, which confers resistance to the herbicide phosphinothricin; the hph gene, which confers resistance to the antibiotic hygromycin; the dhfr gene, which confers resistance to methotrexate; and the EPSPS gene, which confers resistance to glyphosate), chemical resistance marker genes (such as herbicide resistance genes), and mannose-6-phosphate isomerase genes, which provide the ability to metabolize mannose. For the safety of transgenic plants, it is possible to omit any selectable marker genes and directly screen transformed plants using stress.

[0032] In the above applications, the microorganism can be yeast, bacteria, algae, or fungi, such as Agrobacterium. The recombinant microorganism refers to a recombinant microorganism whose genes have been manipulated and modified to produce functionally altered recombinant microorganisms. For example, a recombinant microorganism is obtained by introducing the above-mentioned recombinant vector into the target microorganism. The recombinant microorganism can be understood to refer not only to the specific recombinant microorganism, but also to the progeny of such a cell. Due to natural, accidental, or intentional mutations and / or changes, such progeny may not necessarily be completely identical to the original parent cell, but are still included within the scope of recombinant microorganisms.

[0033] In a third aspect, the present application claims a new use of the above-mentioned MsPIF8 protein or the above-mentioned biological material.

[0034] The present application claims the use of the above-mentioned MsPIF8 protein or the above-mentioned biological material in any one of the following M1) to M10):

[0035] M1) regulating plant growth;

[0036] M2) preparing a product for regulating plant growth;

[0037] M3) regulating plant height;

[0038] M4) preparing a product for regulating plant height;

[0039] M5) regulating the number of nodes of a plant stem;

[0040] M6) preparing a product for regulating the number of nodes of a plant stem;

[0041] M7) cultivating a transgenic plant with increased plant height and / or increased number of nodes of a plant stem;

[0042] M8) preparing a product for cultivating a transgenic plant with increased plant height and / or increased number of nodes of a plant stem;

[0043] M9) plant breeding;

[0044] M10) preparing a product for plant breeding.

[0045] In the above-mentioned uses, the regulating plant growth is promoting plant growth under short-day conditions.

[0046] The regulating plant height is increasing plant height under short-day conditions.

[0047] The regulating the number of nodes of a plant stem is increasing the number of nodes of a plant stem under short-day conditions.

[0048] The index of the plant breeding is plant height or the number of nodes of a plant stem (plant height or the number of nodes of a plant stem under short-day conditions).

[0049] The purpose of the plant breeding is cultivating a plant variety with increased plant height and / or increased number of nodes of a plant stem (cultivating a plant variety with increased plant height and / or increased number of nodes of a plant stem under short-day conditions).

[0050] In a fourth aspect, the present application claims a method for cultivating a transgenic plant with increased plant height and / or increased number of nodes of a plant stem or a method for increasing plant height and / or the number of nodes of a plant stem.

[0051] The method for cultivating transgenic plants with increased plant height and / or increased number of stem nodes claimed in the present invention comprises the following steps: increasing the expression level and / or activity of the above-mentioned MsPIF8 protein in the target plant to obtain a transgenic plant; the plant height and / or number of stem nodes of the transgenic plant are higher than those of the target plant.

[0052] The method for increasing plant height and / or stem node number claimed in the present invention comprises the following steps: increasing the expression level and / or activity of MsPIF8 protein in the target plant to achieve increased plant height and / or stem node number.

[0053] The above-mentioned method for cultivating transgenic plants with increased plant height and / or increased number of stem nodes is a method for cultivating transgenic plants with increased plant height and / or increased number of stem nodes under short-day conditions.

[0054] The above method for increasing plant height and / or stem node number is a method for increasing plant height and / or stem node number under short-day conditions.

[0055] In the above method, the method for increasing the expression level and / or activity of the MsPIF8 protein in the target plant is to overexpress the MsPIF8 protein in the target plant.

[0056] Furthermore, the overexpression method is to introduce the coding gene of the MsPIF8 protein into the target plant.

[0057] Furthermore, the gene encoding the MsPIF8 protein is a DNA molecule shown in SEQ ID NO.1.

[0058] In any of the above applications or methods, the short-day sunshine refers to sunshine time less than (not including or equal to) 12 hours.

[0059] In some embodiments, the short daylight period is 8 hours of daylight and 16 hours of darkness.

[0060] In any of the above-described uses or methods, the transgenic plants include not only first-generation transgenic plants obtained by transforming the target plant with the MsPIF8 gene, but also progeny thereof. Transgenic plants can be propagated within the species or transferred using conventional breeding techniques into other varieties of the same species, particularly commercial varieties. Transgenic plants include seeds, callus, whole plants, and cells.

[0061] In any of the above uses or methods, the plant is a dicotyledonous plant or a monocotyledonous plant.

[0062] Furthermore, the dicotyledonous plant is a leguminous plant.

[0063] Furthermore, the legume plant is a Medicago plant.

[0064] Furthermore, the alfalfa plant is alfalfa.

[0065] In some embodiments, the alfalfa is alfalfa ('Zhongmu No. 1').

[0066] The present invention constructs transgenic alfalfa with increased expression of the MsPIF8 gene. Experiments have shown that, compared with wild-type alfalfa, transgenic alfalfa with increased expression of the MsPIF8 gene exhibits significantly increased plant height and stem node number under short-day conditions. This invention is of great significance for improving alfalfa yield. BRIEF DESCRIPTION OF THE DRAWINGS

[0067] Figure 1 Identification and phenotype of MsPIF8 transgenic alfalfa. a: Schematic diagram of the pMDC83-MsPIF8 vector structure. b: RT-qPCR analysis of MsPIF8 gene expression in MsPIF8 transgenic alfalfa. MsActin was used as an internal reference gene for alfalfa. c: Phenotypes of alfalfa before and after long- and short-day treatments. First row: Phenotypes of alfalfa before long- and short-day treatments; second row: Phenotypes of alfalfa after four weeks of long- and short-day treatments. Scale bar: 5 cm. d: Plant height and number of nodes of alfalfa after four weeks of long- and short-day treatments. DETAILED DESCRIPTION

[0068] The present invention will be further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for further improvements by those skilled in the art and are not intended to limit the present invention in any way.

[0069] Unless otherwise specified, the experimental methods in the following examples are conventional methods and were performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials and reagents used in the following examples, unless otherwise specified, were all commercially available.

[0070] The alfalfa cultivar 'Zhongmu No. 1' in the following examples is described in the document “Shi K, Liu J, Liang H, Dong H, Zhang J, Wei Y, Zhou L, Wang S, Zhu J, Cao M, Jones CS, Ma D, Wang Z. An alfalfaMYB-like transcriptional factor MsMYBH positively regulates alfalfa seedling drought resistance and undergoes MsWAV3-mediated degradation. J Integr Plant Biol. 2024 Apr; 66(4): 683-699.”

[0071] Example 1: Cloning and acquisition of MsPIF8 gene

[0072] 1. Extraction of RNA and cDNA Synthesis from Alfalfa

[0073] Total RNA was extracted from 'Zhongmu No. 1' alfalfa seedlings using the TRIzoL method (Invitrogen, USA). The extracted RNA was reverse transcribed to obtain cDNA using the One-Step gDNA Removal and cDNA Synthesis SuperMix (Cat. No. AT311).

[0074] 2. Using the cDNA obtained in step 1 as a template, amplify using the primer pair T-MsPIF8-F / T-MsPIF8-R to obtain a PCR amplification product. The PCR amplification product is the coding region sequence of the MsPIF8 gene. The primer sequences are as follows:

[0075] T-MsPIF8-F: 5'-ATGAGTCAGTGTGTTCCTCGTTGGGA-3';

[0076] T-MsPIF8-R: 5'-TCAATTCTTTGAACCTGGTGCTGGAG-3'.

[0077] The PCR amplification system was as follows: 2*KOD one 10 μL, T-MsPIF8-F (10 μmol / L) 0.6 μL, T-MsPIF8-R (10 μmol / L) 0.6 μL, and alfalfa cDNA 1 μL, and ddH2O was added to a final volume of 20 μL.

[0078] The PCR reaction program was as follows: pre-denaturation at 94°C for 2 min; denaturation at 98°C for 10 s, extension at 68°C for 20 s, 40 cycles; and 12°C for 5 min.

[0079] 3. The PCR amplification product obtained in step 2 was ligated to the T vector and sequenced. The plasmid with the correct sequence was named T-MsPIF8. The sequencing results showed that the coding region sequence of the MsPIF8 gene is shown in SEQ ID NO. 1, and the amino acid sequence encoded by it is shown in SEQ ID NO. 2, which is the MsPIF8 protein, consisting of 492 amino acid residues.

[0080] Example 2: Preparation of MsPIF8 transgenic alfalfa

[0081] 1. Construction of MsPIF8 overexpression vector pMDC83-MsPIF8

[0082] The plant overexpression vector of MsPIF8 was constructed using Gateway Technology. The specific steps are as follows:

[0083] 1. Design primer pair pDONR207-MsPIF8-F / pDONR207-MsPIF8-R based on the CDS sequence of MsPIF8. The primer sequences are as follows:

[0084] pDONR207-MsPIF8-F:

[0085] 5'-ggggacaagtttgtacaaaaaagcaggcttcATGAGTCAGTGTGTTCCTCGTTG-3';

[0086] pDONR207-MsPIF8-R:

[0087] 5'-ggggaccactttgtacaagaaagctgggtcATTCTTTGAACCTGGTGCTGG-3'.

[0088] 2. Using the T-MsPIF8 plasmid constructed in Example 1 as a template, gene amplification was performed using the primer pair in step 1 to obtain a PCR product, which was named PCR-pDONR207-MsPIF8.

[0089] The PCR amplification system consisted of 10 μL of 2*KOD one, 0.6 μL of p207-MsPIF8-F (10 μmol / L), 0.6 μL of p207-MsPIF8-R (10 μmol / L), and 0.5 μL of T-MsPIF8 plasmid, with ddH2O added to a final volume of 20 μL. The PCR reaction program was as follows: initial denaturation at 94°C for 2 min; 40 cycles of denaturation at 98°C for 10 s, annealing at 56°C for 5 s, and extension at 68°C for 20 s; and storage at 12°C for 5 min.

[0090] 3. After gel recovery of the PCR product pDONR207-MsPIF8 from step 2, it was ligated to the pDONR207 vector using Gateway BP Clonase II (Thermo Fisher). The plasmid that was sequenced correctly was named pDONR207-MsPIF8. The Gateway BP Clonase II ligation system was as follows: 2 μL of PCR product pDONR207-MsPIF8, 0.5 μL of pDONR207 vector, and 0.5 μL of BP enzyme.

[0091] 4. Use Gateway LR Clonase II (Thermo Fisher) to connect the pDONR207-MsPIF8 obtained in step 3 with the pMDC83 vector (NovoPro, V013531). The plasmid with correct sequencing was named pMDC83-MsPIF8. The schematic diagram of the structure of the pMDC83-MsPIF8 plasmid is shown in the figure. Figure 1 As shown in a.

[0092] The Gateway LP ClonaseⅡ ligation system is as follows: 0.5 μL of pDONR207-MsPIF8 plasmid, 2 μL of pMDC83 vector, and 0.5 μL of BP enzyme.

[0093] 2. Acquisition and identification of MsPIF8 transgenic alfalfa

[0094] 1. Transform the recombinant plasmid pMDC83-MsPIF8 constructed in Step 1 into competent Agrobacterium tumefaciens EHA105. After identification, obtain Agrobacterium EHA105 carrying the pMDC83-MsPIF8 recombinant plasmid. Then, use Agrobacterium-mediated transformation of the recombinant plasmid pMDC83-MsPIF8 into leaves of alfalfa 'Zhongmu No. 1', and obtain transgenic seedlings through tissue culture.

[0095] 2. Use the CTAB method to extract the genomic DNA of the transgenic seedlings, and use the primer pair 35S and pMDC83-MsPIF8-R to detect whether the pMDC83-MsPIF8 vector is inserted into the plant. The seedlings containing the correct target band (about 1500bp) are positive seedlings.

[0096] 3. Extract RNA from positive seedlings and leaves from the same part of alfalfa 'Zhongmu No. 1' using the Trizol method and reverse transcribe to obtain cDNA. Use the primer pairs qMsPIF8-F / qMsPIF8-R and MsActin-F / q-MsActin-R, using the extracted cDNA as a template for real-time fluorescence quantitative PCR amplification to detect MsPIF8 expression and screen for transgenic alfalfa with elevated MsPIF8 expression. The primer sequences are as follows:

[0097] 35S: 5'-CGCAAGACCTCCTCTATATAAG-3';

[0098] qMsPIF8-F: 5'-GGCGGTGGATGCGTTGGTTC-3';

[0099] qMsPIF8-R: 5'-CCTCCTTCGTCCTGCGTTGC-3';

[0100] q-MsActin-F: 5'-CAAAAGATGGCAGATGCTGAGGAT-3';

[0101] q-MsActin-R: 5'-CATGACACCAGTATGACGAGGTCG-3'.

[0102] The results are as follows Figure 1 As shown in (b), the results showed that the expression levels of MsPIF8 in the MsPIF8 transgenic alfalfa lines OE-MsPIF8-1, OE-MsPIF8-3, and OE-MsPIF8-10 were significantly higher than those in wild-type alfalfa. The MsPIF8 transgenic alfalfa lines OE-MsPIF8-1, OE-MsPIF8-3, and OE-MsPIF8-10 were selected for the following biological function analysis.

[0103] Example 3: Phenotypic Analysis of MsPIF8 Transgenic Alfalfa

[0104] Test materials: wild-type alfalfa 'Zhongmu No. 1' (CK), MsPIF8 transgenic alfalfa lines OE-MsPIF8-1, OE-MsPIF8-3, and OE-MsPIF8-10.

[0105] The experimental method is as follows:

[0106] (1) Propagation of the test materials by cuttings. After 4 weeks of growth, select the uniformly growing seedlings and transfer them to nutrient soil for further cultivation. After 4 weeks of growth, the seedlings are uniformly cut to a 6 cm stubble to obtain alfalfa plants.

[0107] (2) The alfalfa plants in step (1) were placed in a long-day (light: dark = 16h: 8h) and short-day (light: dark = 8h: 16h) incubators for cultivation at a temperature of 25°C and a light intensity of 200 μmol·m -2 ·s -1 After 4 weeks of growth, the phenotype was observed and the plant height and number of nodes were counted.

[0108] The results are as follows Figure 1 c and Figure 1 As shown in Figure d, the results showed that after 4 weeks of treatment under long-day conditions, the plant height and stem node number of MsPIF8 transgenic alfalfa were no different from those of wild-type alfalfa (CK); however, after 4 weeks of treatment under short-day conditions, the plant height and stem node number of MsPIF8 transgenic alfalfa were significantly increased compared with wild-type alfalfa (CK). Under short-day conditions, the plant heights of wild-type alfalfa (CK) and MsPIF8 transgenic alfalfa lines OE-MsPIF8-1, OE-MsPIF8-3, and OE-MsPIF8-10 were 20.3 cm, 24.2 cm, 24.7 cm, and 25.2 cm, respectively. The number of nodes in wild-type alfalfa (CK) and MsPIF8 transgenic alfalfa lines OE-MsPIF8-1, OE-MsPIF8-3, and OE-MsPIF8-10 were 6.4, 8, 9, and 8.3, respectively. This suggests that the MsPIF8 gene can regulate plant growth under short-day conditions.

[0109] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be practiced over a wide range of parameters, concentrations, and conditions without departing from the spirit and scope of the present invention and without unnecessary experimentation. Although specific embodiments have been given herein, it should be understood that further modifications may be made to the present invention. In summary, this application is intended to encompass any variations, uses, or improvements to the present invention, including those made by conventional techniques known in the art that depart from the scope of the present invention. Applications of the essential features may be made within the scope of the following claims.

Claims

1. A protein, which is a protein represented by a) or b) or c) or d) below: a) the amino acid sequence is the protein shown in SEQ ID NO. 2; b) a fusion protein obtained by connecting a tag to the N-terminus and / or C-terminus of the protein shown in SEQ ID NO. 2; c) a protein having the same function 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; d) A protein having 75% or more identity with the amino acid sequence shown in SEQ ID NO. 2 and having the same function.

2. The biological material related to the protein according to claim 1, which is any one of the following A1) to A8): A1) a nucleic acid molecule encoding the protein according to claim 1; A2) an expression cassette containing the nucleic acid molecule described in A1); A3) a recombinant vector containing the nucleic acid molecule described in A1); A4) a recombinant vector containing the expression cassette described in A2); A5) a recombinant microorganism containing the nucleic acid molecule described in A1); A6) a recombinant microorganism containing the expression cassette described in A2); A7) a recombinant microorganism containing the recombinant vector described in A3); A8) A recombinant microorganism containing the recombinant vector described in A4).

3. The biomaterial according to claim 2, characterized in that: A1) The nucleic acid molecule is the gene shown in 1) or 2) below: 1) The coding sequence is the DNA molecule shown in SEQ ID NO.1; 2) A DNA molecule that has 75% or more identity with the nucleotide sequence defined in 1) and encodes the protein of claim 1.

4. Use of the protein according to claim 1 or the biomaterial according to claim 2 or 3 in any of the following M1) to M10): M1) Regulate plant growth; M2) preparing products for regulating plant growth; M3) regulating plant height; M4) preparing products for regulating plant height; M5) Regulate the number of plant stem nodes; M6) preparing products for regulating plant node number; M7) cultivating transgenic plants with increased plant height and / or increased number of stem nodes; M8) preparing products for cultivating transgenic plants with increased plant height and / or increased number of nodes; M9) Plant breeding; M10) Preparing products of plant breeding.

5. The use according to claim 4, characterized in that: The regulating plant growth is to promote plant growth under short-day conditions; Alternatively, the regulating plant height is to increase the plant height under short-day conditions; Alternatively, the regulating the number of plant nodes is to increase the number of plant nodes under short-day conditions.

6. The use according to claim 4 or 5, characterized in that: The plant is a dicotyledonous plant or a monocotyledonous plant.

7. A method for cultivating a transgenic plant having increased plant height and / or increased number of stem nodes, comprising the steps of: increasing the expression level and / or activity of the protein of claim 1 in a target plant to obtain a transgenic plant; wherein the plant height and / or number of stem nodes of the transgenic plant are greater than those of the target plant; Alternatively, a method for increasing plant height and / or stem node number comprises the following steps: increasing the expression level and / or activity of the protein according to claim 1 in the target plant to achieve increased plant height and / or stem node number.

8. The method according to claim 7, wherein: The method for cultivating transgenic plants with increased plant height and / or increased stem node number is a method for cultivating transgenic plants with increased plant height and / or increased stem node number under short-day conditions; Alternatively, the method for increasing plant height and / or stem node number is a method for increasing plant height and / or stem node number under short-day conditions.

9. The method according to claim 7 or 8, characterized in that: The method for increasing the expression level and / or activity of the protein according to claim 1 in the target plant is to overexpress the protein according to claim 1 in the target plant; Alternatively, the overexpression method is to introduce the gene encoding the protein according to claim 1 into the target plant.

10. The method according to any one of claims 7 to 9, characterized in that: The plant is a monocotyledonous plant or a dicotyledonous plant.

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