CN116813734A - OsEF1A protein, encoding gene and application thereof in promoting rice tillering - Google Patents

Abstract

The application discloses an OsEF1A protein, a coding gene and application thereof in promoting rice tillering, and belongs to the technical field of genetic engineering. The OsEF1A protein can obviously improve the tillering number of a Nipponbare rice plant, further improves the rice yield, provides a new way for breeding new varieties of high-yield rice, provides a new gene resource and recombinant plasmid preparation method, and has important significance in facilitating the yield and income increase of rice.

Description

OsEF1A protein, encoding gene and its application in promoting rice tillering

Technical field

The present invention relates to the technical field of genetic engineering, in particular to OsEF1A protein, encoding genes and their application in promoting rice tillering.

Background technique

Rice (Oryza sativa L.), commonly known as rice, is an annual aquatic herb of the family Gramineae (perennial rice varieties already exist). The stalks are upright, 0.5-1.5 meters high, depending on the variety. The leaf sheath is hairless and flaccid; the ligule is lanceolate; the blade is linear-lanceolate, about 1 cm wide, hairless and rough. The panicle is large and sparsely spread, with rough edges; the spikelet contains 1 mature flower; the globule is extremely small, leaving only a half-moon mark on the apex of the spikelet stalk, which is cone-like; the pregnant lemma on both sides is thick and has 5 veins. , the midrib is ridged, with small checkered papillary protrusions on the surface, thick paper, fine hairs all over and dense terminal hairs, with or without awns; the lemma is the same as the lemma, with 3 veins, apex is sharp but without beak; stamens Anthers are 2-3 mm long. The caryopsis is about 5 mm long and 2 mm wide; the embryo ratio is about 1/4 of the caryopsis length.

The number of tillers is one of the most important agronomic traits of rice, because the number of tillers directly determines the number of panicles per plant, which in turn affects rice yield. The occurrence of rice tillering can be divided into two steps: the formation of axillary buds and the elongation of axillary buds. Currently, cloned genes controlling axillary bud formation include MOC1, MOC3/TAB1, TAD1/TE, LAX1, LAX2, etc. MOC1 is the first cloned gene that controls rice tillering. It encodes a GRAS family transcription factor and affects tillering by controlling axillary bud formation. The MOC1 mutant has only one main stem and does not produce tillers. Strigolactones (SLs) are a new class of plant hormones that are involved in regulating plant branching and root development, and interacting with symbiotic arbuscular mycorrhizal fungi and parasitic weeds. SLs signaling inhibits rice tiller bud elongation. Related genes that catalyze the synthesis of SLs (such as D27, D17, D10 and OsMAX1) and signal transduction (such as D3, D14, D53, IPA1) are involved in the regulation of rice tillering. Among them, IPA1 is a semi-dominant gene encoding OsSPL14 transcription factor, and its expression is regulated by microRNA OsmiR156. The point mutation of this gene hinders the function of OsmiR156, resulting in an "ideal plant type" with reduced tiller number, enhanced lodging resistance, and increased yield. Recent studies have shown that IPA1 can interact with D53 and thereby function as a downstream transcription factor in the SL signaling pathway. Although multiple genes controlling rice tillering have been cloned and their regulatory mechanisms have been basically elucidated, new genes in the rice tillering regulatory network still need to be identified.

Contents of the invention

The purpose of the present invention is to provide OsEF1A protein, encoding genes and their application in promoting rice tillering.

In order to achieve the above objects, the present invention provides the following solutions:

The present invention provides an OsEF1A protein, the amino acid sequence of the OsEF1A protein is shown in SEQ ID NO: 2.

The present invention also provides DNA molecules encoding the OsEF1A protein.

Preferably, the nucleotide sequence of the DNA molecule is shown in SEQ ID NO: 1.

The invention also provides recombinant vectors, expression cassettes, transgenic cell lines or recombinant bacteria containing the DNA molecules.

Preferably, the recombinant vector includes pCAMBIA1300S-2×35S::OsEF1A.

The present invention also provides primer pairs for amplifying the DNA molecules.

Preferably, the primer pair includes an upstream primer as shown in SEQ ID NO: 3 and a downstream primer as shown in SEQ ID NO: 4.

The present invention also provides the application of the OsEF1A protein, the DNA molecule or the recombinant vector, expression cassette, transgenic cell line or recombinant bacteria in promoting rice tillering.

The present invention also provides a method for cultivating multi-tiller rice, which is characterized in that the DNA molecule is introduced into the recipient rice.

Preferably, the DNA molecule is introduced into the target plant through the recombinant vector, expression cassette, transgenic cell line or recombinant bacteria.

The invention discloses the following technical effects:

The present invention provides OsEF1A protein, encoding genes and their application in promoting rice tillering. The OsEF1A protein can significantly increase the number of tillers of Nipponbare rice plants, thereby increasing rice yield, providing a new approach to the breeding of new high-yielding rice varieties, and providing A new genetic resource and recombinant plasmid preparation method have been developed, which is of great significance for increasing rice production and income.

Description of the drawings

In order to explain the embodiments of the present invention or the technical solutions in the prior art more clearly, the drawings needed to be used in the embodiments will be briefly introduced below. Obviously, the drawings in the following description are only some of the drawings of the present invention. Embodiments, for those of ordinary skill in the art, other drawings can also be obtained based on these drawings without exerting creative efforts.

Figure 1 shows the PCR identification of positive transgenic plants; where, M: DNA quality standard; CK: negative control;

Figure 2 shows the tissue expression of OsEF1A in rice, GUS staining of roots, stems, rhizome junctions, and spikelets of PBI121 OsEF1Apro::GUS-positive plants;

Figure 3 shows the subcellular localization of OsEF1A;

Figure 4 is a phenotypic diagram of transgenic plants;

Figure 5 shows the different expression levels of OsEF1A gene detected by fluorescence quantification.

Detailed ways

Various exemplary embodiments of the invention will now be described in detail. This detailed description should not be construed as limitations of the invention, but rather as a more detailed description of certain aspects, features and embodiments of the invention.

It should be understood that the terms used in the present invention are only used to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges in the present invention, it should be understood that every intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or value intermediate within a stated range and any other stated value or value intermediate within a stated range is also included within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded from the range.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only the preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and/or materials in connection with which the documents relate. In the event of conflict with any incorporated document, the contents of this specification shall prevail.

It will be apparent to those skilled in the art that various modifications and changes can be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to the skilled person from the description of the invention. The specification and examples are intended to be illustrative only.

The words "includes", "includes", "has", "contains", etc. used in this article are all open terms, which mean including but not limited to.

The technical solutions described in the present invention, unless otherwise specified, are all routine solutions in the field. The reagents or raw materials used, unless otherwise specified, are all purchased from commercial channels or have been disclosed.

Example 1

1. Rice RNA extraction

(1) Grind Nipponbare rice leaves in liquid nitrogen, add 450 μL RL (add 1% β-mercaptoethanol), and vortex vigorously to mix.

(2) Transfer all solutions to the filter column CS, centrifuge at 12,000 rpm for 3 minutes, and carefully draw the supernatant into an RNase-free centrifuge tube.

(3) Slowly add 0.5 times the volume of supernatant absolute ethanol, mix well, transfer all solutions to the adsorption column CR3, centrifuge at 12000 rpm for 60 seconds, and pour out the waste liquid in the collection tube.

(4) Add 350 μL of protein-removing solution RW1 to the adsorption column CR3, centrifuge at 12,000 rpm for 60 seconds, and discard the waste liquid in the collection tube.

(5) Add 80 μL of DNaseⅠ working solution (10 μL DNaseⅠ storage solution + 70 μL RDD buffer) to the center of the adsorption column CR3, and leave it at room temperature for 15 minutes.

(6) Add 350 μL of protein-removing solution RW1 to the adsorption column CR3, centrifuge at 12,000 rpm for 60 seconds, and discard the waste liquid in the collection tube.

(7) Add 500 μL of rinse solution RW to the adsorption column CR3, let it stand at room temperature for 2 minutes, centrifuge at 12000 rpm for 60 seconds, and discard the waste liquid in the collection tube.

(8) Repeat step 7.

(9) Centrifuge at 12,000 rpm for 2 minutes and discard the waste liquid. Place the adsorption column CR3 at room temperature for a few minutes to completely dry the residual rinse solution in the adsorption column.

(10) Place adsorption column CR3 into a new RNase-Free centrifuge tube, drop 50 μL RNase-Free ddH ₂ O into the center of the adsorption column, and centrifuge at 12,000 rpm for 2 minutes to obtain an RNA solution.

2. Reverse transcription of cDNA

(1) According to Table 1, prepare the reaction system on ice, with a total volume of 10 μL.

Table 1

(2) Vortex to mix, and centrifuge briefly.

(3) Incubate at 42°C for 2 minutes.

(4) After the reaction is completed, cool it on ice.

(5) Prepare the reaction system according to Table 2, with a total volume of 20 μL.

Table 2

(6) Mix well and centrifuge briefly.

(7) Incubate at 42°C for 30 minutes and 85°C for 5 minutes to obtain a cDNA solution.

3. Primer design

Search the NCBI database to find the rice Nipponbare genome and obtain the rice OsEF1A gene sequence. The OsEF1A gene sequence is shown in SEQ ID NO: 1.

SEQ ID NO: 1:

cccgtcctctccttcaagtcagctacctgcttaatcaaccatgggtaaggagaagacgcacatcaacattgtggtcattggccacgtcgactctggcaagtcgaccaccacgggccatctgatctacaagcttggaggtatcgacaagcgtgtgattgagaggttcgagaaggaagccgctgagatgaacaagaggtccttcaagt acgcgtgggtgctcgacaagctcaaggccgagcgtgagagaggtatcaccatcgatattgccctgtggaagttcgagaccaccaagtactactgcacggtcatcgatgcccctggtcaccgtgacttcatcaagaacatgatcacgggtacctcccaggctgactgtgccgtgctcatcattgactccaccactggtggtttt gaggctggtatctccaaggatggccagaccgtgagcacgctcttcttgctttcactcttggtgtgaagcagatgatctgctgctgcaacaagatggatgccaccactcccaagtactccaaggcccgttacgatgaaatcgtgaaggaagtctcatcctacctgaagaaggtcggctacaaccctgacaagattcccttcgtt cccatctctgggtttgagggtgacaacatgattgagaggtccaccaaccttgactggtacaagggccccaccttgcttgaggctcttgaccagatcaacgagcccaagaggccatcagacaagcccctgcgtcttccccttcaggacgtgtacaagatcggtggtattggcaccgtgcctgtgggtcgtgttgagactggagtcct caagcctggtatggtggtgacctttggtcccagcggcctgaccactgaggtcaagtcggttgagatgcaccacgaggctctccaggaggctcttcctggtgacaacgtcgggttcaacgtgaagaacgttgcggtgaaggatctgaagcgtgggtacgtggcctccaactccaaggatgaccctgccaaggaggctgcc agcttcacctcccaggtgatcatcatgaaccaccctggccagatcggcaacggctacgccccagtgctggactgccacacctcccacattgccgtcaagtttgctgagctggtgaccaagatcgacagacgatctggtaaggagctggagaaggagcccaagttcctcaagaacggtgatgctggtatggttaagatgattcccaccaagcccatggtt gtggagaccttctctgagtaccctcctcttggtcgttttgccgtgcgtgacatgaggcaaacggtggctgttggcgtcatcaagaacgtggagaagaaggacccaaccggtgccaaggtgaccaaggctgccgccaaagaagaaatgatcctcgttcaagtggtagttttgtgcacggccatggcactcgttg cattgaccgttattatcatatgctcgctgtgtgcactgtgtcgtttaaaactctctattttctgttcggtttaaacttgttctgagcctgattattgcccttcggctgttgtggaatgcgggtatttacattgctaattcctatgtctttattttggcttgtggcattcgcttgttctactgctattgaaaact gtaatctgtttctgttattaaaaactgaaccttttcttctgctatttgaaactgaagcgagtgggtttcaatttgctgaaatcctgctacttgctatttaaattatttatgttcaaaaaaaaaaaaaaaaaa;

The protein sequence encoded by the OsEF1A gene is shown in SEQ ID NO: 2.

SEQ ID NO: 2:

MGKEKTHINIVVIGHVDSGKSTTTGHLIYKLGGIDKRVIERFEKEAAEMNKRSFKYAWVLDKLKAERERGITIDIALWKFETTKYYCTVIDAPGHRDFIKNMITGTSQADCAVLIIDSTTGGFEAGISKDGQTREHALLAFTLGVKQMICCCNKMDATTPKYSKARYDEIVKEVSSYLKKVGYNPDKIPFVPISGFEGDNMIERSTNLDWYKGPTL LEALDQINEPKRPSDKPLRLPLQDVYKIGGIGTVPVGRVETGVLKPGMVVTFGPSGLTTEVKSVEMHHEALQEALPGDNVGFNVKNVAVKDLKRGYVASNSKDDPAKEAASFTSQVIIMNHPGQIGNGYAPVLDCHTSHIAVKFAELVTKIDRRSGKELEKEPKFLKNGDAGMVKMIPTKPMVVETFSEYPPLGRFAVRDMRQTVAVGVIKNVEKK DPTGAKVTKAAAKKK.

Design primers based on the restriction site of vector pCAMBIA1300S. The primers are as follows:

1300-OsEF1A-BamHI-F:ATGGGTAAGGAGAAGACGCACATCA;

1300-OsEF1A-PstI-R: AGCGTAATCTGGAACATCGTATGGGTA;

Hyg-1300-301-F (SEQ ID NO: 3): CAAGACCTGCCTGAAACGAACT;

Hyg-1300-741-R (SEQ ID NO: 4): GGCCGTCTGCTGCTCCATACA.

4. Vector construction

Methods include PCR amplification of target genes, purification of PCR products, extraction of E. coli plasmids, enzyme digestion, ligation, transformation of E. coli, PCR identification of positive colonies, identification of plasmid enzyme digestion, sequencing and strain preservation, etc.

Use PCR technology to clone the CDS of LRK2 from cDNA. The primers are the same as 3. Primer design. The gene fragment and expression vector were double digested with BamHI and PstI respectively, and the digested products were recovered by gel cutting respectively. The recovered products were subjected to ligation, transformation, positive clone identification, sequencing and sequence analysis, and finally the expression vector: pCAMBIA1300S-2×35S::OsEF1A was obtained.

5. Glue recycling

SanPrep Column DNA Gel Recovery Kit operates step by step for purification.

(1) Cut the gel piece of the target fragment from the agarose gel and weigh it.

(2) Add B2 B2 that is 5 times the weight of the gel block, and sol it in a 50°C water bath for 10 minutes.

(3) Transfer the solution to the adsorption column and centrifuge at 8000 rpm for 30 seconds.

(4) Add 500 μL Wash SoLution and centrifuge at 9000 rpm for 30 seconds.

(5) Repeat step (4).

(6) Centrifuge the empty adsorption column at 9000 rpm for 1 minute.

(7) Place the adsorption column into a clean 1.5 mL centrifuge tube, add 20 μLElutionBuffer to the center of the adsorption column, let stand at room temperature for 1 min, and centrifuge at 12,000 rpm for 1 min to obtain a DNA solution.

6.Connect

Generally, ligation is performed overnight at 4°C. The ligation reaction system is as shown in Table 3. The volume of the reaction system (10 μL in total):

table 3

7. Preparation of E. coli competent cells

(1) Take the strain, streak it on LB medium, and culture it at 37°C overnight.

(2) Pick a single colony on the LB plate, inoculate it into about 5 mL of LB liquid culture medium, and culture it overnight at 37°C and 250 rpm.

(3) Inoculate the bacterial solution into 50 mL LB liquid medium at a ratio of 1:50, and culture it at 37°C and 250 rpm for 1-2 hours until the OD ₆₀₀ of the bacterial solution is between 0.5-0.6.

(4) Transfer the bacterial solution into a 50mL centrifuge tube and pre-cool for 30 minutes.

(5) Centrifuge at 4°C and 4000rpm for 10 minutes. Discard supernatant.

(6) Add 5mL SSCS solution to suspend the cells.

(7) Dispense into 1.5mL centrifuge tubes (cool in advance), 100μL per tube.

(8) Quick freeze in liquid nitrogen, then transfer to -80°C for storage.

8. E. coli transformation

(1) Take out the competent state from -80℃, melt it on ice, add the ligation product and mix gently with a gun, and keep in ice bath for 30 minutes.

(2) Heat shock at 42°C for 90 seconds and immediately transfer to ice. Ice bath for 10 minutes.

(3) Add 700 μL LB liquid medium and incubate at 37°C and 180 rpm for 1 hour.

(4) Apply on LB solid culture medium (containing antibiotics) and blow dry on a clean bench.

(5) Place upside down in a 37°C biochemical incubator and incubate overnight.

9. Positive clone identification and sequencing

(1)PCR identification

Use Taq Plus DNA Polymerase for PCR identification. The reaction system is shown in Table 4. The total volume is 10 μL. (Pick a single colony from the transformed plate, number it, gently touch a single colony with a 10 μL pipette tip, put it in a PCR tube and stir it gently to use as a template).

Table 4

The reaction procedure is shown in Table 5:

table 5

(2)Send for testing

The constructed vector was sent to the sequencing department of Shanghai Yingjun Biotechnology Co., Ltd. for sequencing.

10. Plasmid DNA extraction

(1) Add 1-5 mL of the bacterial liquid cultured overnight into a 1.5 mL centrifuge tube, centrifuge at 13,000 rpm for 30 seconds, collect the bacterial sediment, and discard the supernatant.

(2) Add 250 μL Buffer P1 to the centrifuge tube containing the bacterial sediment, mix thoroughly with a vortex shaker, and suspend the bacterial sediment.

(3) Add 250 μL Buffer P2 to the centrifuge tube, mix gently by inverting up and down 4-6 times, and mix thoroughly to lyse the bacteria.

(4) Add 350 μL Buffer N3 to the centrifuge tube, immediately mix gently by inverting up and down 8-10 times, and mix thoroughly. A white flocculent precipitate should appear at this time. Centrifuge at 13,000rpm for 5min.

(5) Transfer the supernatant obtained in step 4 to the adsorption column installed in the collection tube, centrifuge at 13,000 rpm for 30 seconds, pour out the waste liquid in the collection tube, and put the adsorption column back into the collection tube.

(6) Add 150 μL Buffer PB to the adsorption column and centrifuge at 13,000 rpm for 30 seconds.

(7) Add 400 μL Buffer PW to the adsorption column, centrifuge at 13,000 rpm for 1 min, and discard the waste liquid in the collection tube.

(8) Place the adsorption column in a new centrifuge tube, add 50 μL Buffer EB to the middle of the adsorption membrane, leave it at room temperature for 2 minutes, and centrifuge at 13,000 rpm for 1 min to obtain a plasmid DNA solution.

11. Preparation of Agrobacterium competent cells

(1) Take out the GV3101 strain at -80°C, streak it on the LB medium plate supplemented with 50 mg/mL Rif, and culture it at 28°C for 2 days.

(2) Pick 1-10 single clones, inoculate them into 250 mL of LB liquid medium containing 50 mg/mL Rif, and culture at 28°C until OD ₆₀₀ = 0.5.

(3) Transfer the bacterial solution to a 1.5mL centrifuge tube and place it on ice for 30 minutes.

(4) Centrifuge at 5000rpm for 5 minutes and discard the supernatant.

(5) Add 750 μL of pre-cooled 0.15M/L NaCl to gently suspend the cells and place on ice for 20 minutes.

(6) Centrifuge at 5000rpm for 5 minutes and discard the supernatant.

(7) Add 100 μL of pre-cooled 20mM/L _CaCl2 solution containing 15% glycerol and gently suspend the cells.

(8) Quick freeze in liquid nitrogen and store at -80°C.

12. Agrobacterium heat shock transformation

(1) Take the competent Agrobacterium stored at -80°C and place it in an ice bath for 20 minutes until it thaws.

(2) Add 5 μL of positive plasmid to the competent cells, mix gently by pipetting, and place on ice for 30 minutes.

(3) Freeze the competent state in liquid nitrogen for 3 minutes, quickly place it in a 37°C water bath for 5 minutes, take it out and place it on ice for 5 minutes.

(4) Add 600 μL of LB liquid culture medium and place on a shaker at 28°C for recovery, 250 rpm, for 1 hour.

(5) Evenly spread the bacterial solution on the solid medium containing Rif and Kan resistance.

(6) Place the coated culture medium in a 28°C incubator and incubate it upside down for 48 hours.

13.Construction of OsEF1A gene overexpression rice lines

(1) Obtaining transgenic rice:

①Rice receptor: Nipponbare;

②Required culture medium (liquid):

a. Medium for callus induction (pH value 5.8): N6D+2mg/L 2,4-D;

b. Agrobacterium infection solution (pH value 5.2): AAM+10-20mg/L AS;

c. Medium for callus co-culture after infection (pH value 5.2): N6D+2mg/L 2,4-D+10-20mg/L AS;

d. Screening medium (pH value 5.8): N6D+2mg/L 2,4-D+500mg/L Cef+100-150mg/L corresponding antibiotic (G418 or Kan);

e. Differentiation medium (pH value 5.8): RE-III+0.02mg/L NAA+2mg/L KT+125mg/LCef+70-100mg/L corresponding antibiotic (G418 or Kan);

f. Rooting medium: HF+125mg/L Cef+70mg/L corresponding antibiotic (G418 or Kan); pH value 5.8;

③Operating steps:

Ⅰ. Induction of rice callus.

Ⅱ. Agrobacterium infection.

Ⅲ. Co-culture of rice callus.

Ⅳ.Debacterization and screening of resistant calli.

Ⅴ. Differentiation of resistant callus.

Ⅵ. Rooting and hardening of differentiated seedlings.

(2) Positive identification of transgenic plants

Follow the Taq Plus DNA Polymerase instructions.

The plasmid vector pCambia1300 hygromycin phosphotransferase gene (Hyg-1300-301-F and Hyg-1300-741-R) was used to conduct PCR identification using tissue culture regenerated seedling leaf tissue DNA as a template. The negative control (CK) was Nipponbare DNA. The reaction system is shown in Table 6:

Table 6

The reaction procedure is shown in Table 7:

Table 7

After the reaction, the results of the gel electrophoresis detection experiment are shown in Figure 1.

14.OsEF1A promoter expression analysis

The PBI121 OsEF1A pro::GUS vector contains the GUS reporter gene. If the tissue parts (roots, stems, leaves, sheaths, etc.) of the transgenic plant can be stained blue by the GUS dye, it means that the plant is positive. The preparation of GUS dye solution is shown in Table 8. Immerse the specific rice tissue in the GUS staining solution, vacuum it for 15 minutes, and leave it at 37°C overnight; remove the GUS staining solution, and use a decolorizing solution (V _ethanol : V _{acetic acid} = 3:1) to wash away the background color of the rice plant material. Use OLYMPUS SZX16 stereomicroscope to observe and take pictures as shown in Figure 2, and store at 4°C.

Table 8 1×GUS dye solution formula

15. Subcellular localization of OsEF1A

(1) Fusion expression vector of OsEF1A and CFP genes, and introduce 35S::CFP:OsEF1A into onion epidermal cells using biolistic method.

(2)Cultivation and observation

After the injection is completed, the plants are returned to the incubator. After 2 days of regular cultivation, the leaves can be cut and observed with a laser scanning confocal microscope.

The results are shown in Figure 3. Subcellular localization expression of OsEF1A gene expression was observed by laser confocal microscopy and showed that it was mainly located in the cell membrane, cytoskeleton and nucleus.

16. Two OsEF1A gene overexpression lines, OE-1 and OE-4, were genetically constructed. Nipponbare and transgenic plants OE-1 and OE-4 were planted respectively. The routine management is shown in Figure 4. The different expression levels of OsEF1A gene were detected by fluorescence quantification, and the detection results are shown in Figure 5. Count and record the number of tillers as shown in Table 9.

Table 9 Tiller number statistics

Note: The NIP strain is Nipponbare, and the OE-1 strain and OE-4 strain are OsEF1A overexpression strains.

As shown in Table 9, the tiller number of transgenic Nipponbare rice plants overexpressing the OsEF1A gene is significantly higher than that of Nipponbare, thereby increasing rice yield, providing a new way for the breeding of new high-yielding rice varieties, and providing a new genetic resource and recombination Plasmid preparation methods are of great significance for increasing rice production and income.

The above-described embodiments only describe preferred modes of the present invention and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, those of ordinary skill in the art can make various modifications to the technical solutions of the present invention. All deformations and improvements shall fall within the protection scope determined by the claims of the present invention.

Claims (9)

1. An OsEF1A protein, characterized in that the amino acid sequence of the OsEF1A protein is shown in SEQ ID NO: 2. 2. A DNA molecule encoding the OsEF1A protein of claim 1. 3. The DNA molecule according to claim 2, wherein the nucleotide sequence of the DNA molecule is as shown in SEQ ID NO: 1. 4. Recombinant vector, expression cassette, transgenic cell line or recombinant bacteria containing the DNA molecule of any one of claims 2-3. 5. A primer pair for amplifying the DNA molecule according to any one of claims 2-3. 6. The primer pair according to claim 5, characterized in that the primer pair includes an upstream primer as shown in SEQ ID NO: 3 and a downstream primer as shown in SEQ ID NO: 4. 7. Application of the OsEF1A protein of claim 1, the DNA molecule of any one of claims 2-3, or the recombinant vector, expression cassette, transgenic cell line or recombinant bacterium of claim 4 in promoting rice tillering. 8. A method for cultivating multi-tiller rice, which is characterized in that the DNA molecule described in any one of claims 2-3 is introduced into the recipient rice. 9. The method according to claim 8, characterized in that the DNA molecule of any one of claims 2-3 is introduced into the target plant through the recombinant vector, expression cassette, transgenic cell line or recombinant bacterium of claim 4.