Moso bamboo PeMASR1 gene, biological material and application thereof

Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a moso bamboo PeMASR1 gene, a biological material and application of the moso bamboo PeMASR1 gene. The PeMASR1 gene provided by the invention is a new gene found in moso bamboo leaf cold resistance research, the nucleotide sequence of the PeMASR1 gene is as shown in SEQ ID NO: 1, and the amino acid sequence of the coded protein is as shown in SEQ ID NO: 2. Through gene clone expression and comparative analysis, the phyllostachys edulis PeMASR1 gene phylloevolution status and the important effect of plants on abiotic stress mechanism research and photosynthesis are disclosed. Overexpression in a Columbia type arabidopsis thaliana plant enables rosette leaves of the plant to age premature; the arabidopsis thaliana shows a phenotype which is more stress-resistant than wild arabidopsis thaliana in coping with abiotic stress such as cold stress, drought stress and salt stress.

Description

Technical Field

[0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a bamboo PeMASR1 gene, a biological material and applications thereof. Background Art

[0002] bamboo( Phyllostachys edulis ) is a large herbaceous plant of the subfamily Bambusoideae of the Poaceae family (Li et al. (2018), Plant Biotechnology Journal l16(1):72–85; Wang et al. (2017), Plant Journal 91(4):684–699). It has important ecological, cultural and economic significance and is the most important economic bamboo species in my country, supporting about 70% of the output value of my country's bamboo industry (Peng et al. (2013), Plos One 8(11):e78944). The current drastic changes in the global environment have led to frequent natural disasters such as heat damage, salt damage and drought. For example, in 2013, the continuous high temperature and low rainfall in southern China caused a large number of bamboo shoots to die, resulting in a sharp decline in its yield (Zhang et al. (2022), Plant Physiology and Biochemistry 186:121–134.). Similarly, soil salinization seriously affects the growth and geographical distribution of bamboo (Gao et al. (2019), Journal of Plant Growth Regulation 38(3):1127–1140; Hou et al. (2020), Tree Physiology 40(12):1792–1806). Researchers have found that even short-term treatment with 150mmol / L NaCl did not allow one-year-old bamboo seedlings to survive (Xu et al. (2021), Plant Cell Reports 40(10):1971–1987). Therefore, it is very important to explore the molecular mechanism of bamboo stress resistance and identify key resistance genes for molecular breeding of new bamboo germplasm that is resistant to stress.

[0003] At the same time, as a representative bamboo species of temperate bamboo, moso bamboo can adapt to the winter temperatures of minus to 10 degrees Celsius in southeastern my country. ◦ C, and has high cold tolerance. Wang et al. (2022) (Tree Physiology 42(11):2336–2352) used a cold-sensitive bamboo species, Bamboo of the Small Buddha Belly ( Bambusa ventricosa) was used as a control, and the adaptive mechanism of bamboo leaves to low temperatures in winter was explored from the perspectives of anatomy, physiology, metabolomics and transcriptomics. It was found that compared with the leaves of Phyllostachys edulis, the anatomical structures related to cold stress resistance of bamboo leaves were similar to those of Phyllostachys edulis, such as fewer spindle cells, smaller bubble cells, lower water content and stomatal density, but more epidermal hairs; non-targeted metabolomics analysis of the leaves of the two bamboos showed that under warm and cold conditions in winter, the anti-stress metabolites such as glutathione, trehalose and ascorbic acid in bamboo leaves were 10 to 1000 times higher than those in Phyllostachys edulis leaves; compared with warm conditions in winter, under cold conditions in winter, Metabolites such as glutathione and trehalose in the leaves of Moso bamboo further increased under the conditions of warm and cold days in winter, while those in the leaves of Phyllostachys edulis decreased significantly; comparative analysis of the transcriptome of Moso bamboo leaves under warm and cold days in winter showed that metabolism-related genes such as trehalose, glutathione, flavonoid metabolism, DNA repair, and reactive oxygen degradation, as well as hormone-related genes such as jasmonic acid, abscisic acid, and ethylene constituted the molecular network of Moso bamboo leaves in response to cold stress; among various transcription factors, HSF, MYB, NAC, and WRYK transcription factors in Moso bamboo leaves were significantly upregulated under low temperature; in addition, a large number of newly expressed Moso bamboo genes were identified to be involved in the regulatory network. It can be seen that Moso bamboo leaves have evolved special anatomical structures, metabolic pathways, and cold stress response molecular regulatory networks to adapt to the cold environment in winter. However, there are currently no reports on key genes for cold resistance in Moso bamboo leaves.

[0004] PeMASR1 ( M ultiple A biotic S tress R The esistant 1) gene is a new gene discovered in the study of the cold tolerance mechanism of bamboo leaves. Its expression is extremely significantly upregulated in bamboo leaves on cold days in winter, and its expression level is nearly 30 times that of bamboo leaves on warm days in winter (Wang et al. (2022), Tree Physiology 42(11):2336–2352). However, there are currently no reports on the functions of this gene and its orthologous genes in other plants. Summary of the invention

[0005] In view of the shortcomings of the prior art, the present invention provides a bamboo PeMASR1 Genes, biomaterials and their applications can be used to study the molecular mechanisms of plant leaf aging, leaf growth and plant stress resistance and can be further applied in plant breeding and improvement, playing an important role in plant genetic engineering such as improving plant resistance to adverse stress.

[0006] The technical solution provided by the present invention is as follows:

[0007] The invention provides a kind of moso bamboo PeMASR1A gene encoding the following protein (a) or (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 2; (b) A protein derived from (a) with equivalent functions, wherein one or more amino acids are substituted, deleted or added to the sequence shown in SEQ ID NO: 2.

[0008] Furthermore, bamboo PeMASR1 The nucleotide sequence of the gene is shown in SEQ ID NO: 1. The present invention clones the gene using the following method: PeMASR1 Gene:

[0009] (1) Extracting total RNA from bamboo leaves In the present invention, the total RNA from bamboo can be extracted by a method commonly used in the art for extracting total cellular RNA, for example, the Novagen FastPure Universal Plant Total RNA Isolation Kit can be used to extract total RNA.

[0010] (2) Reverse transcription of the extracted total RNA from bamboo to synthesize cDNA. In the present invention, conventional cDNA synthesis methods in the art can be used, for example, Tiangen (Beijing) One-Step cDNA Synthesis Kit can be used to synthesize cDNA.

[0011] (3) After obtaining cDNA, PeMASR1 PCR amplification of the gene to obtain the target fragment. PeMASR1 The nucleotide sequences of the primers for the genes are as follows: PeMASR1 -F: 5'-ATGGGGGATGTGTCGTTG-3' PeMASR1 -R:5'-TTATGTTGCTTGATCATTGCT-3'.

[0012] (4) After PCR amplification to obtain the target fragment, the target fragment is sequenced to obtain PeMASR1 Gene.

[0013] (5) The target fragment was connected to the pCAMBIA1302 vector and introduced into E. coli DH5α competent cells. After verification as a positive clone by colony PCR, sequencing was performed.

[0014] This invention also provides bamboo PeMASR1 The biomaterial of the gene is any one or more of recombinant DNA, expression cassette, transposon, plasmid vector, viral vector, engineered bacteria or non-renewable plant cells or tissues.

[0015] The present invention also provides a bamboo PeMASR1The application of a gene or a biological material containing the gene in promoting plant premature senescence, regulating plant leaf growth and / or regulating plant stress resistance (improving plant stress resistance); wherein the stress resistance refers to drought resistance, salt resistance and cold resistance; and the plant is Arabidopsis thaliana.

[0016] The present invention also provides a bamboo PeMASR1 The application of the gene or biological material containing the gene in preparing transgenic plants. The breeding purpose is to improve the stress resistance of plants.

[0017] The present invention also provides a method for improving the cold resistance, salt resistance and / or drought resistance of plants, comprising: using genetic engineering means to overexpress the gene of claim 1 in the plant; the overexpression method is selected from any one or more combinations of the following (1)-(5): (1) by introducing a plasmid containing the gene; (2) by increasing the copy number of the gene on the plant chromosome; (3) by changing the promoter sequence of the gene on the plant chromosome; (4) by operably linking a strong promoter to the gene; (5) by introducing enhancers; The plant is Arabidopsis thaliana. In the present invention, the expression vector carrying the target gene can be introduced into plant cells by conventional biotechnology methods such as Ti plasmid, plant virus vector, direct DNA transformation, microinjection, electroporation, etc.

[0018] Furthermore, the bamboo PeMASR1 The gene was transferred into Arabidopsis plants to obtain overexpression of bamboo PeMASR1 Genetically modified plants.

[0019] Furthermore, the bamboo PeMASR1 The gene was constructed into the plant expression vector pCAMBIA1302, transformed into Agrobacterium, and then infected into Arabidopsis inflorescence to screen transgenic plants.

[0020] The present invention also provides the use of the transgenic plants obtained by the above method in plant breeding.

[0021] Furthermore, breeding methods include transgenic, hybridization, backcrossing, selfing or asexual reproduction.

[0022] Beneficial Effects

[0023] The present invention discloses for the first time PeMASR1 The biological function of genes. PeMASR1The invention provides a method for using the gene to encode the evolutionary status of the gene system, and overexpression of the gene can make the Columbia type Arabidopsis plants more tolerant to cold, salt and drought. PeMASR1 Genes will play an important role in plant response to abiotic stress. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 for PeMASR1 Gel electrophoresis diagram of gene amplification products.

[0025] Figure 2 for PeMASR1 Amino acid sequence alignment of the gene with rice Os10g25060 gene.

[0026] Figure 3 for PeMASR1 Rosette leaf morphology of transgenic plants and Col-0 plants at 28 days of growth. PeMASR1 The leaves of transgenic plants (OE10 and OE11) showed obvious senescence compared with the control plants (Col-0).

[0027] Figure 4 The phenotype photos before and after cold treatment, drought treatment and salt treatment, where Col-0 represents the wild-type plant of Arabidopsis Columbia, and OE10 and OE11 represent different lines of transgenic plants.

[0028] Figure 5 for PeMASR1 Phenotypic diagram of transgenic Arabidopsis and wild-type Arabidopsis plants under cold stress, where Col-0 represents the wild-type plant of Arabidopsis Columbia, and OE10 and OE11 represent different strains of transgenic plants, respectively.

[0029] Figure 6 for PeMASR1 Phenotypic diagram of transgenic Arabidopsis and wild-type Arabidopsis plants under drought stress, where Col-0 represents the wild-type plant of Arabidopsis Columbia, and OE10 and OE11 represent different strains of transgenic plants, respectively.

[0030] Figure 7 for PeMASR1 Phenotypic diagram of transgenic Arabidopsis and wild-type Arabidopsis plants under salt stress, where Col-0 represents the wild-type plant of Arabidopsis Columbia, and OE10 and OE11 represent different strains of transgenic plants, respectively. DETAILED DESCRIPTION

[0031] The present invention is further described below in conjunction with specific examples. Molecular biology experimental methods not specifically described in the following examples are all performed with reference to the specific methods listed in the book Molecular Cloning Experiment Guide (3rd Edition) by J. Sambrook, or performed according to the kits and product instructions.

[0032] Example 1: Bamboo PeMASR1 Obtaining gene sequences

[0033] 1. RNA extraction

[0034] The extraction material was bamboo leaves. RNA was extracted using the Novagen FastPure Universal Plant Total RNA Isolation Kit according to the operating instructions and stored at -80°C for future use.

[0035] 2. First-strand cDNA synthesis and reverse transcription PCR

[0036] Total RNA was reverse transcribed into cDNA using Tiangen (Beijing) One-Step cDNA Synthesis Kit according to the operating instructions. The reaction system and reaction conditions were: 1µg of prepared total RNA, 1µL of Anchored Oligo (dT) 18 Primer, 10µL of 2×TS Reaction Mix, 1µL of TransScript RT / RI Enzyme Mix, 1µL of gDNA Remover, and RNase-free Water was added to 20µL, gently mixed and centrifuged. Reverse transcription was performed at 42℃ for 30min, 85℃ for 5s; cooled on ice; stored at -20℃ after centrifugation for use.

[0037] 3. PCR amplification PeMASR1 CDS area

[0038] The gene sequence was obtained based on the transcriptome sequencing results of bamboo leaves. Two primers were designed upstream and downstream of its CDS region. PeMASR1 -F(5'-ATGGGGGATGTGTCGTTG-3') and PeMASR1 -R (5'-TTAT GTTGCTTGATCATTTGCT-3') was used as the primer for the PCR reaction.

[0039] The PCR enzyme used was the high-fidelity enzyme P525 from Nanjing Novozyme for PCR amplification. The reaction system was 20 µL: ddH2O, 9.5 µL; 2xPhantaMaxMasterMix (DyePlus) enzyme, 12.5 µL; forward primer, 1 µL; reverse primer, 1 µL; cDNA, 1 µL. The PCR program was: 95°C for 3 min; 95°C for 15 s, 56°C for 15 s, 72°C for 36 s, 35 cycles; 72°C for 5 min. The resulting PCR product was separated by 1.2% agarose gel electrophoresis to obtain a 597 bp fragment (CDS length 597 bp), such as Figure 1 After purification and recovery, the DNA was sent to Qingke Biotechnology Co., Ltd. (Nanjing) for sequencing.

[0040] Example 2: Bamboo PeMASR1 Gene function analysis

[0041] 1. Sequence comparative analysis

[0042] Analyzed with DNAMAN8 PeMASR1 The gene has similarity to its closest relative, Os10g25060, in terms of amino acid sequence. Figure 2 As shown, PeMASR1 The sequence similarity between the gene and Os10g25060 is 66.83%.

[0043] 2. Arabidopsis transgenic analysis

[0044] 2.1 Vector construction

[0045] Vector linearization: Use endonucleases NcoI and BstEII to double-digest the vector. The reaction system is: pCAMBIA1302 vector, 10 μL; NcoI, 1 μL; BstEII, 1 μL; rcut smart, 5 μL; ddH2O, 33 μL.

[0046] 2.2 Overexpression vector construction: Use C115 homologous recombinase from Norvegic for homologous recombination, the reaction system is: pCAMBIA1302 linearized vector, 1 μL; DNA, 1.5 μL; 2×ClonExpreesMix, 2.5 μL. Use a pipette to gently mix, the reaction program is 50°C, 5 min.

[0047] 2.3 Transformation: Take 50 μL of competent cells DH5α and place them on ice to melt, add 5 μL of the above-mentioned connected DNA, flick to mix, and place on ice for 30 minutes; heat shock in a 42°C water bath for 45 seconds, and immediately place on ice for 2 minutes; add 900 μL of room temperature LB medium without antibiotics, and culture at 200 rpm and 37°C for 1 hour; centrifuge at 5000 rpm for 5 minutes to obtain bacteria, remove most of the supernatant, and use the remaining culture medium to resuspend the bacteria. Use a sterile spreader to gently spread and seal the LB plate containing kanamycin; invert and culture in a 37°C incubator for 12-16 hours.

[0048] 2.4 Bacterial liquid PCR: Add 0.5mL LB culture medium to a 1.5mL EP tube, select 5 colonies of appropriate size on the plate, mark 1-5 with a pen, and do not place the colonies too close to each other. Clip out 5 sterile enzyme-free centrifuge tubes in advance, and pipette 2×RapidTaqMasterMix, 10μL; ddH2O, 7µL; vector upstream primer, 1µL; vector downstream primer, 1µL; use a toothpick to pick up a single clone and place the colony into the centrifuge tube. The colony PCR program is: 95℃ 3min, 95℃ 15s, 56℃ 15s, 72℃ 9s, 35 cycles; 72℃ 5min. The obtained colony PCR product is subjected to 1.2% agarose gel electrophoresis, and the remaining colonies with the correct length band are picked into a 500µL test tube containing kanamycin LB medium, placed on a shaker for 4h, and then sent to Qingke Biotechnology Co., Ltd. for sequencing. Sequencing results and PeMASR1 The plasmids were extracted from bacterial liquid with consistent sequences to be transformed into Agrobacterium.

[0049] 3 Arabidopsis transformation

[0050] 3.1 Seeding

[0051] Take peat soil, vermiculite, and perlite and mix them evenly in a ratio of 8:4:1, and add dimethoate (about 3-4 spoons per basket of soil). Put the soil into a small flower pot, water the bottom (add 1g / L of Huawuque for the first watering), and sow when the soil in the pot absorbs water and becomes moist. Take the seeds of wild-type Colombia Col-0 and put them in a 4℃ refrigerator for about three days, take them out and transplant them into the soil, cover them with plastic wrap, and put them in an artificial climate box for cultivation at a temperature of 24℃ and a light of 120UM. After about 2 days, remove the plastic wrap and cultivate them until the flowering period.

[0052] 3.2 Agrobacterium transformation

[0053] Take the competent Agrobacterium stored at -80℃, melt it by hand, and insert it into ice when it is in an ice-water mixing state; add 5μL of the constructed plasmid containing the target gene to every 50μL of competent state, gently stir the bottom of the centrifuge tube to mix, let it stand on ice for 5min, liquid nitrogen for 5min, 37℃ water bath for 5min, and ice bath for 5min; add 700μL of antibiotic-free LB liquid culture medium, and culture at 28℃ with shaking for 2~3h; centrifuge at 6000rpm for 1min, remove most of the supernatant, keep about 100μL of supernatant, gently blow to resuspend the bacteria, apply LB plates containing kanamycin and rifampicin, and invert and culture for 2~3d.

[0054] Pick a single Agrobacterium colony for colony PCR, pick the bacteria with the correct band and shake it for one day at 28°C, 200 rpm, until the OD value is about 0.8~1; centrifuge at 4500 rpm for 10 minutes, remove the supernatant, resuspend the bacteria with sucrose suspension (5% sucrose and 0.03% surfactant Silwet), and adjust the OD 600 The value reaches 0.8~1.0 and activated under light for 2~3h.

[0055] 3.3 Arabidopsis transformation by inflorescence dip method

[0056] Take 3 to 4 pots of Arabidopsis thaliana in full flowering stage, cut off the fruit pods and blooming flowers in advance, immerse the inflorescence in the activated suspension for about 1 minute, culture in a humid dark environment for 24 hours, then take out and place in an incubator for normal culture.

[0057] 3.4 Screening, observation and identification

[0058] 1) When the siliques of Arabidopsis thaliana turn yellow or brown, bag the plants and stop watering to allow them to slowly die, then harvest them uniformly to obtain transgenic T0 seeds; the harvested T0 seeds are placed in a 4°C refrigerator for low-temperature treatment for about 3 days; 1 / 2MS solid culture medium is prepared, sterilized by high pressure, and then placed in an ultra-clean workbench to slowly cool down. When the culture medium temperature drops to about 50°C, hygromycin is added at a concentration of 20 mg / L. At the same time, prepare 1 / 2MS culture medium without hygromycin for later use; take an appropriate amount of transgenic T0 seeds after vernalization, disinfect with 75% ethanol for 5 minutes, then use a cut gun tip to suck in anhydrous ethanol to blow and wash the seeds, sprinkle on sterilized filter paper, and after the anhydrous ethanol evaporates, evenly sprinkle the seeds on the 1 / 2MS plate containing hygromycin. At the same time, spread Col-0 seeds on the 1 / 2MS culture medium without resistance as a control; put the plate into an incubator for incubation, and when the seedlings on the resistant 1 / 2MS culture medium take root and the first pair of true leaves unfold, transplant the seedlings to the prepared soil, water them sufficiently, cover them with plastic wrap for 2-3 days, and then remove the plastic wrap.

[0059] 2) Take 1-2 rosette leaves from transgenic positive seedlings of different strains and use the Novagen FastPure kit to extract RNA. The operation steps refer to the instruction manual. All consumables such as centrifuge tubes and pipette tips are RNase-free.

[0060] a. Grind 1-2 leaves in liquid nitrogen, add 600 μL Buffer EL, vortex vigorously for 30 seconds to mix the sample and lysate thoroughly, centrifuge at 12000 rpm for 5 minutes, and proceed with subsequent operations immediately.

[0061] b. Take about 500 μL of the supernatant and transfer it to FastPuregDNA-FilterColumnsⅢ, centrifuge at 12000 rpm for 30 seconds, discard FastPuregDNA-FilterColumnsⅢ, and collect the filtrate.

[0062] c. Add 0.5 times the filtrate (about 250 μL) to the collection tube and vortex to mix for 15 seconds.

[0063] d. Transfer the above mixed solution to FastPureRNA Columns V, centrifuge at 12000 rpm for 30 seconds, and discard the filtrate.

[0064] e. Add 700 μL Buffer RWA to FastPure RNA Columns V, centrifuge at 12000 rpm for 30 s, and discard the filtrate.

[0065] f. Add 500 μL Buffer RWB to Fast Pure RNA Columns V (please check whether 48 mL of anhydrous ethanol has been added before use), centrifuge at 12000 rpm for 30 s, and discard the filtrate.

[0066] g. Repeat step 6.

[0067] h. Place FastPureRNA Columns V back into the collection tube and centrifuge at 12000 rpm for 2 min.

[0068] i. Transfer FastPureRNAColumnsV to new RNase-freeCollectionTubes1.5ml centrifuge tubes, add 40μL RNase-freeddH2O in the center of the adsorption membrane and centrifuge at 12000rpm for 1min (the first eluate can be added back to the adsorption column for elution to improve the recovery efficiency).

[0069] j.1.2% agarose gel electrophoresis was used to detect the integrity of the extracted RNA.

[0070] 3) Using qRT-PCR technology, and Arabidopsis ACTIN1 Gene as internal reference to identify transgenic plants PeMASR1 The reverse transcription method was similar to the first-strand cDNA synthesis and reverse transcription PCR in Example 1, and the cDNA was diluted 10 times for later use. The qRT-PCR enzyme was ChamQ TM UniversalSYBR, system is 20μL: H2O, 7.2μL; Mix, 10μL; Upstream / Downstream Primers: 0.4μL; cDNA Template, 2μL. Set the program to pre-denaturation 95℃ 30s; 95℃ 10s, 60℃ 30s, 40 cycles; 95℃ 15s, 60℃ 60s, 95℃ 15s. Specific primers are as follows: PeMASR1 -F:5'-TACCAAGCAGGGCGATCAAA-3', PeMASR1 -R: 5'-ACCGGAATTGCGTGGTCTAA-3'; AtActin-F :5'-CTCTCCCGCTTTGAATTGTCTCGTTG-3', AtActin-R :5'-GGTACCATTGTCACACACGATTGGT-3'.

[0071] Income PeMASR1 Transgenic Arabidopsis thaliana OE10 and OE11 lines PeMASR1 The relative expression levels of 175.7 and 238.2 were respectively; its phenotype showed early senescence of leaves and smaller leaves ( Figure 3 ).

[0072] 3.5 Cold stress, drought stress and salt stress in transgenic plants

[0073] 3.5.1 Cold stress treatment

[0074] The treated material was three-week-old Arabidopsis thaliana, which was placed in a low-temperature incubator at 4°C. After four days, it was obvious that the leaves of the wild-type Col-0 Arabidopsis thaliana plants wilted and died. PeMASR1 Transgenic plants OE10 and OE11 have good growth conditions (see Figure 5 ).

[0075] 3.5.2 Drought stress treatment

[0076] The treated material was three-week-old Arabidopsis. After 10 days of watering cessation, the four-week-old transgenic strain entered the senescence stage and the leaves turned yellow. After 14 days of drought, it was obvious that the leaves of the wild-type Col-0 Arabidopsis Columbia plants wilted and turned yellow, and the overall state was worse than that of OE10 and OE11 (see Figure 6 ).

[0077] 3.5.3 Salt stress treatment

[0078] The treated material was three-week-old Arabidopsis thaliana, which was treated with 200 mM / L NaCl solution by root irrigation. After 6 days of treatment, it was clearly observed that the four-week-old transgenic strain entered the early senescence stage, and the leaves turned yellow. After 16 days of salt treatment, the leaves of the wild-type Col-0 Arabidopsis thaliana wilted and turned yellow, and the overall state was worse than that of OE10 and OE11 (see Figure 7 ).

[0079] Figure 4 This is the growth status of Arabidopsis before cold treatment, drought treatment and salt treatment.

[0080] SEQ ID NO: 1: .

[0081] SEQ ID NO:2: MGDVSLNQPINAEPLPCSVTKQGDQILDLMSAGWTNERHSLYISSMEASFMEQLYGHEHHGPDANWSHVGGNGFKVLQEGVSENLRIKRNDAHARDGGVICLPDNPWIRRFRPRNSGINRHGDGVRASVDDGESGTDTVQERVRTHGREVKSCVGENLAEVCDQNFPDEDVKANAEASKSCKKRRPTLSTAAANDQAT。

Claims

1. A PeMASR1 gene of bamboo, characterized in that: It is a gene encoding the following protein (a) or (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 2; (b) A protein derived from (a) with equivalent functions, wherein one or more amino acids are substituted, deleted or added to the sequence shown in SEQ ID NO:

2.

2. The gene according to claim 1, characterized in that Its nucleotide sequence is shown in SEQ ID NO:

1.

3. A biological material containing the gene according to claim 1 or 2, characterized in that: The biological material is any one or more of recombinant DNA, expression cassette, transposon, plasmid vector, virus vector, engineered bacteria or non-renewable plant cells or tissues.

4. Use of the gene according to claim 1 or 2 or the biomaterial according to claim 3 in promoting plant premature senescence, regulating plant leaf growth and / or regulating plant stress resistance; wherein, The stress resistance refers to drought resistance, salt resistance and cold resistance; the plant is Arabidopsis thaliana.

5. Use of the gene according to claim 1 or 2 or the biological material according to claim 4 in preparing transgenic plants.

6. A method for improving the cold resistance, salt resistance and / or drought resistance of plants, characterized in that: include: Using genetic engineering methods to overexpress the gene of claim 1 in plants; The overexpression method is selected from any one or more combinations of the following (1)-(5): (1) by introducing a plasmid containing the gene; (2) by increasing the copy number of the gene on the plant chromosome; (3) by changing the promoter sequence of the gene on the plant chromosome; (4) by operably linking a strong promoter to the gene; (5) by introducing enhancers; The plant is Arabidopsis thaliana.

7. The method according to claim 6, characterized in that The bamboo PeMASR1 gene was transferred into Arabidopsis plants to obtain transgenic plants overexpressing the bamboo PeMASR1 gene.

8. The method according to claim 7, characterized in that The PeMASR1 gene of moso bamboo was constructed into the plant expression vector pCAMBIA1302, transformed into Agrobacterium, and then infected into Arabidopsis thaliana inflorescence to screen transgenic plants.

9. Use of the transgenic plant obtained by the method according to any one of claims 6 to 8 in plant breeding.

10. The use according to claim 9, characterized in that: Breeding methods include transgenic, hybridization, backcrossing, selfing or asexual reproduction.

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