CN109081865B

CN109081865B - Phyllostachys pubescens PeVQ28 protein and coding gene and application thereof

Info

Publication number: CN109081865B
Application number: CN201811056089.8A
Authority: CN
Inventors: 项艳; 程欣然; 王玉娇; 熊瑞; 吴敏; 严涵薇
Original assignee: Anhui Agricultural University AHAU
Current assignee: Anhui Agricultural University AHAU
Priority date: 2018-09-11
Filing date: 2018-09-11
Publication date: 2021-08-10
Anticipated expiration: 2038-09-11
Also published as: CN109081865A

Abstract

The invention discloses a moso bamboo PeVQ28 protein, and a coding gene and application thereof. The amino acid sequence of the protein is shown as SEQ ID No.1, the coding gene sequence (CDS) is shown as SEQ ID No.2, and the full length of the cDNA sequence of the Phyllostachys pubescens PeVQ28 gene is shown as SEQ ID No. 3. The PeVQ28 gene provided by the invention has high expression under the induction of salt stress of different degrees. The PeVQ28 gene expression vector is transferred into wild type Arabidopsis thaliana by an agrobacterium-mediated inflorescence dip-dyeing method, and the result shows that the salt resistance of the transgenic wild type Arabidopsis thaliana is obviously improved, which indicates that the PeVQ28 gene can improve the salt resistance of the transgenic Arabidopsis thaliana. The plant material obtained by the moso bamboo PeVQ28 gene expression vector has certain adaptability to the external salt stress environment and has good application prospect.

Description

Phyllostachys pubescens PeVQ28 protein and coding gene and application thereof

Technical Field

The invention relates to the field of plant molecular biology, and in particular relates to a moso bamboo PeVQ28 protein, and a coding gene and application thereof.

Background

The variety of Chinese bamboos is very rich, about 39 types and 500 types are counted, the category source and the planting area of the bamboos and the storage amount and the yield of bamboo forests are all the first in the world, and China is the most important world for producing the bamboos. The moso bamboo is also a forest resource, occupies a very important position in southern areas of China, can replace some slow-growing forest resource types due to the high growth speed, short period reaching available standards and considerable yield, and has wider application range for the moso bamboo along with the development of science and technology. Due to the nature and the characteristics of the moso bamboo, the use of the bamboo on buildings gradually attracts operators in scenic spots and farmhouses, and the bamboo building has unique shape, simplicity, elegance, low energy and environmental protection; bamboo-made bamboo products such as besoms, chairs, soup spoons, toys and the like gradually enter the daily life of people; the high cellulose content of moso bamboo determines its usefulness as a paper making raw material. Meanwhile, in the aspect of eating, not only the bamboo shoots cannot be eaten as green food materials, but also the components in green drinks or health care products processed by some factories are extracted from bamboo leaves or hydrolyzed and refined from bamboo processing waste materials. Besides economic value, the moso bamboo forest also has important ecological value, such as conserving water sources, conserving soil, purifying environment and the like. .

VQ-motif proteins are a class of plant specific proteins that are of increasing interest due to interactions with WRKY transcription factors. Through research on a VQ protein family in Arabidopsis thaliana, the VQ protein family is involved in function regulation of plants in various aspects and mainly involved in salt stress reaction of the plants. Research in Arabidopsis shows that AtCaMBP25(AtVQ15) can negatively regulate osmotic stress in the early germination and growth development stage of Arabidopsis seeds, and the transgenic strain line shows high sensitivity in the germination and growth development of the seeds under the NaCl and osmotic stress conditions. Similarly, it was found that the AtVQ9 overexpression plants show inhibition of seed germination and growth of Arabidopsis seedlings under NaCl treatment.

The growth environment of the moso bamboo has high requirements on moisture, climate and soil, and is suitable for growing in the environment with sufficient moisture, warm climate and slightly acidic or neutral loam. In recent years, with the increasing global temperature and the worsening of climate, plants are harmed by various adverse conditions from the environment during the growth process, and the salinization of soil is a common concern in the world today. Salt stress is one of the major limiting factors in plant growth and development. The moso bamboos are sensitive to salt stress, and when the moso bamboos grow in saline-alkali soil, due to the stress action of salt and alkali, the moso bamboos grow slowly, and often leaves turn yellow, die and fall off, so that photosynthesis is seriously influenced, and sometimes even the whole plant withers and dies. The negative effects of salt stress on moso bamboo are mainly shown in the following aspects: nutritional stress caused by insufficient intake of potassium, calcium, phosphorus and nitrogen; ionic poisoning effects caused by the accumulation of sodium ions, chloride ions, and sulfate; osmotic stress caused by high concentrations of solutes in the soil; reactive oxygen species accumulation causes destruction of biological membranes, proteins and nucleic acids, i.e., oxidative stress. The growth conditions of the moso bamboos are greatly limited by environmental stresses such as high salt and the like, so that the reaction mechanism of the plants to the salt stress is known, the salt resistance of the plants is improved, a batch of new salt-resistant moso bamboo germplasms are cultivated, and the method has important significance for improving the yield of the moso bamboos, promoting the stable development of forest industry and increasing the economic income of the nation.

The VQ gene and its homologous genes published at present are mostly derived from model plants or herbaceous plants, which are annual plants, and the function of the VQ gene in moso bamboo has not been reported yet.

Disclosure of Invention

In order to solve the problems in the prior art, the invention aims to provide a moso bamboo PeVQ28 protein, and a coding gene and application thereof.

In order to realize the purpose of the invention, the invention adopts the following technical scheme: the amino acid sequence of the moso bamboo PeVQ28 protein is shown as SEQ ID No. 1.

The invention relates to a coding gene sequence for coding the moso bamboo PeVQ28 protein, which is characterized in that: the nucleic acid sequence of the moso bamboo PeVQ28 protein is shown in SEQ ID No. 2.

The vector of the invention contains the coding gene sequence.

The construction method of the vector comprises the following steps:

a moso bamboo PeVQ28 protein coding gene sequence with two ends containing restriction enzyme cutting sites is constructed on a pEASY T1 single Cloning Vector, and is transferred into an escherichia coli competent cell TransT1 to obtain a T1-PeVQ28 recombinant plasmid, a double restriction enzyme T1-PeVQ28 recombinant plasmid and a pCAMBIA1301a plasmid, after connection, the recombinant plasmid is transferred into an escherichia coli competent cell, and after the plasmid is extracted, an expression Vector pCAMBIA1301a-PeVQ28 is obtained.

The engineering bacterium containing the carrier is disclosed.

The primer pair for cloning the coding gene sequence is characterized in that: the primer pair comprises an upstream primer and a downstream primer, wherein the nucleotide sequence of the upstream primer is shown as SEQ ID No.4, and the nucleotide sequence of the downstream primer is shown as SEQ ID No. 5.

The invention discloses a moso bamboo PeVQ28 gene, which is characterized in that: the full-length nucleic acid sequence of the Phyllostachys pubescens PeVQ28 gene cDNA is shown as SEQID No. 3.

The moso bamboo PeVQ28 gene is applied to modification of plant salt resistance.

Further, the gene or the coding gene sequence is transferred into a plant by an agrobacterium-mediated inflorescence dip dyeing method.

Has the advantages that: the invention provides a PeVQ28 protein related to salt resistance of moso bamboo for the first time, and a coding gene and application thereof. The PeVQ28 gene expression vector is transferred into wild type Arabidopsis thaliana by an agrobacterium-mediated inflorescence dip-dyeing method, and the result shows that the salt resistance of an over-expressed Arabidopsis thaliana plant is obviously enhanced, which indicates that the PeVQ28 gene can improve the salt resistance of transgenic Arabidopsis thaliana. The plant material obtained by the moso bamboo PeVQ28 gene expression vector has certain adaptability to the external high-salt environment and has good application prospect.

Drawings

FIG. 1 is an amino acid sequence and domain partition diagram of a protein encoded by the PeVQ28 gene according to the present invention;

FIG. 2 is an electrophoretogram of total RNA of Phyllostachys pubescens leaves in example 2 of the present invention; m: trans2K DNA Marker; 1: total RNA of moso bamboo leaves.

FIG. 3 is a schematic diagram of the construction of a plant transgenic vector pCAMBI1301a-PeVQ28 of the PeVQ28 gene of Phyllostachys pubescens of the invention example 4;

FIG. 4 is a phenotypic view of transgenic Arabidopsis thaliana and wild type Arabidopsis thaliana before and after salt treatment according to example 5 of the present invention; a is a phenotype diagram of transgenic arabidopsis and wild arabidopsis before and after salt treatment; b: survival rates of transgenic and wild type arabidopsis thaliana after 10 days of salt treatment; C-H: PRO, RWC, SOD, POD, CAT, and MDA contents of transgenic Arabidopsis and wild type Arabidopsis before and after salt treatment. WT: wild type Arabidopsis thaliana; l1, L3, L10: transgenic overexpression lines.

Detailed Description

The invention is further described with reference to the accompanying drawings and specific embodiments. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

The reagents used in the examples mainly include molecular biology experimental reagents, kits, and the like, which are commercially available, and specifically include: the RNA extraction kit is purchased from Roche Biotechnology Ltd; a reverse transcription Kit (First-Strand cDNA Synthesis Kit) was purchased from Promega corporation; SYBR Green Mix is available from ABI, USA; trizol is purchased from agarose gel DNA recovery kit of Shanghai Invitrogen biotechnology, Inc., and a plasmid rapid miniprep kit is purchased from Axygen; pEASY T1simple Cloning Vector, Taq enzyme, Trans T1 competence and related kits were purchased from Beijing Quanyujin; restriction enzymes SmaI and Sal I and DNAmarker were purchased from TaKaRa (Takara Bio Inc.); sequencing work was done by Huada Gene science and technology, Inc. The methods provided in the examples of the present invention are conventional methods unless otherwise specified.

Example 1

Protein sequence coded by moso bamboo PeVQ28 gene

The full length cDNA sequence of the Phyllostachys pubescens PeVQ28 gene was translated (SEQ ID No.1) using primers software (FIG. 1).

Example 2

Phyllostachys pubescens salt-resistant related gene PeVQ28 induced expression pattern analysis

Stress treatment and Total RNA extraction

Annual bamboo seedlings were irrigated with 100mM sodium chloride solution, 150mM sodium chloride solution and 200mM sodium chloride solution, respectively, keeping other nutritional conditions unchanged. The treated moso bamboo leaves of 0, 1, 3, 6, 12 and 24 hours are taken out by scissors (cleaned and disinfected by 75% alcohol in advance) respectively, put into a 2mL dof tube containing RNA protective solution and stored for standby, and the plants are repeated for 3 times in different time periods and samples. The Trizol method is used for extracting total RNA and comprises the following detailed steps:

(1) taking out the standby moso bamboo leaves from the RNA protective solution, putting the moso bamboo leaves in filter paper to suck the protective solution as dry as possible to avoid influence, transferring the moso bamboo leaves into a mortar, pouring liquid nitrogen into the mortar, immediately grinding the moso bamboo leaves until the moso bamboo leaves become powder, and collecting the powder in a 1.5mLdorf tube (precooling the moso bamboo leaves with the liquid nitrogen in advance);

(2) sucking 1mL of Trizol solution by using a pipettor, adding the Trizol solution into a dof tube filled with a sample, quickly shaking the dof tube for 15s, and then placing the dof tube on an ultraclean workbench for 5min at room temperature;

(3) centrifuge at 4 ℃ for 10min, set at 12,000 rpm. After the end, carefully sucking the middle aqueous phase of the three liquid phases in the 800 mu L dof tube into a new dof tube;

(4) adding chloroform/isoamylol solution 200 μ L, shaking rapidly for 15s, standing for 3 min;

(5) centrifuging at 4 deg.C for 15min at 12,000rpm, sucking 400 μ L of upper water phase into a new dof tube, adding isopropanol with the same volume for precipitation, standing for more than 25 min;

(6) repeating the centrifugal setting of the step (5), and only leaving the sediment at the bottom of the dof tube after finishing;

(7) washing the precipitate with 75% alcohol twice, rotating at 7500rpm, centrifuging for 5min, and collecting the precipitate;

(8) and (4) placing the uncovered dof tube on an ultra-clean workbench for drying, and keeping the room temperature for 5-10 min. The drying time cannot be too long, otherwise, the precipitate is too dry and is difficult to dissolve;

(9) dissolving the precipitate with 30 μ L DEPC water;

(10) after mixing uniformly, obtaining RNA solution, taking out 4 microliter of detection result, and storing the rest in a refrigerator at-80 ℃.

(II) detection of Total RNA and treatment of genome-free DNA

(1) Total RNA integrity assay

And (3) mixing 3 mu L of extracted total RNA with 1 mu L of loading buffer, carrying out electrophoresis detection, and carrying out electrophoresis at 5V/cm. After 20 minutes, the extracted RNA was considered not degraded if the 28S rRNA band near the spotted well was found to be approximately twice as bright as the 18S rRNA band, intact and free of bands, observed in a gel imaging system and photographed (FIG. 2).

(2) Detection of Total RNA purity and content

The purity and content of total RNA were determined by a microspectrophotometer, the instrument was cleaned and RNA (1 μ L) was detected. If the ratio of A260/A280 is in the range of 1.8-2.0, the RNA purity is better, if the RNA is polluted by protein or phenol, the ratio of A260/A280 is obviously lower than 1.8, and if the ratio is more than 2.0, the RNA sample is degraded.

(3) Removal of genomic DNA

1) The following reaction solutions were prepared in a PCR tube:

in a microcentrifuge tube, the following reagents were added in sequence:

RNase-free water was replenished to a final volume of 50. mu.L

2) Reacting at 37 ℃ for 30 minutes, and adding 50 mu L of RNase-free water;

3) 100. mu.L of phenol/chloroform/isoamyl alcohol mixture was aspirated (ratio 25: 24: 1) adding;

4) centrifugation (10000rpm, 10 min), transfer of the aqueous layer to a fresh PCR tube, addition of 10. mu.L sodium acetate (3mol/L, pH 5.2) and 250. mu.L absolute ethanol (precooling), and incubation at-20 ℃ for 45 min;

5) washing the centrifuged precipitate with pre-cooled 70% ethanol, blow-drying on an ultra-clean bench, dissolving the precipitate with RNase-free water, and detecting whether the genomic DNA is removed by electrophoresis.

(III) reverse transcription

Experimental method Using Takara reverse transcription kit PrimeScript^TM RT Master Mix(Perfect Real Time)

The specification provides:

(1) sample adding:

(2) after sample adding and mixing, incubation is carried out for 15 minutes at 37 ℃, and the time can be prolonged to 30 minutes if the RNA concentration is too low;

(3) 5 seconds at 85 ℃ in order to inactivate the reverse transcriptase and preserve at 4 ℃;

(4) the reverse transcription product cDNA was stored in a-20 ℃ freezer for fluorescent quantitation experiments.

(IV) primer design

Fluorescence quantification was designed using Primer 5 software based on the full length of cDNA sequence of the Phyllostachys pubescens PeVQ28 gene

PeVQ28 primer:

F：5’-TGAGCCCATCAACTACACTGCC-3’；

R：5’-TGCTGGCGTCTGGCTTCC-3’。

the specificity of the PCR is detected by using PCR, and the PCR kit can be used on the premise of ensuring the specific amplification of the PCR. The Phyllostachys pubescens TIP41 gene is used as a positive control of a fluorescence quantitative test, and primers of the TIP41 gene are as follows:

F：5’-AAAATCATTGTAGGCCATTGTCG-3’；

R：5-ACTAAATTAAGCCAGCGGGAGTG-3’。

the fluorescent quantitative PCR reaction system is 25 mu L, each component is SYBR Green Mix 12.5 mu L, the template cDNA is 2 mu L, the upstream primer and the downstream primer (10 mu mol/L) are 0.5 mu L respectively, and finally, deionized water is supplemented to 25 mu L. The PCR reaction parameters were as follows: pre-denaturation at 95 ℃ for 10 min; 95 ℃ for 15sec, 60 ℃ for 1min, for 40 cycles. And after the reaction is finished, heating the product to obtain the dissolution curve of the product. By using 2^–ΔΔCTThe method processes the acquired signals and data.

Example 3

Cloning of salt-resistant related gene PeVQ28 of moso bamboo

According to a PeVQ28 gene CDS sequence published on a moso bamboo genome website, a primer for PCR amplification of the fragment is designed, wherein a Sma I restriction enzyme site is added to an upstream primer, and a Sal I restriction enzyme site is added to a downstream primer.

The primer sequences are as follows:

PeVQ28-F5′-ATACCCGGGATGGGGGAGTACCACAG-3′

PeVQ28-R5-GCCGTCGACTTAAGATGCGTACATTTCAC-3′

performing PCR amplification by using a moso bamboo leaf totalRNA reverse transcription 1 st chain cDNA as a template and using an upstream primer and a downstream primer, wherein a PCR reaction system is shown in a table 1:

TABLE 1

The PCR reaction conditions are as follows: pre-denaturation: 10min at 98 ℃; denaturation: 10s at 98 ℃; annealing: 30s at 58 ℃; extension: 1min at 72 ℃ and 39 cycles; total extension: 10min at 72 ℃.

After the reaction is finished, sucking the PCR product for 2% agarose gel electrophoresis, detecting and cutting the gel in a gel imaging system, recovering by using an agarose gel DNA recovery kit, connecting the recovered fragment with pEASY T1 single Cloning Vector, and transforming into Escherichia coli competence TransT1 cells of the all-in-one gold biotechnology Limited company; PCR and enzyme digestion detection screening positive clones; and (3) preliminarily judging the correct connecting fragment according to the detection result, sending the Huada corporation to perform sequencing, wherein the sequencing result is as follows: the DNA sequence of SEQ ID NO.2 in the sequence table is shown and consists of 747bp bases, and the result is completely consistent when the DNA sequence is compared with the sequence reported on a phyllostachys pubescens genome website.

Example 4

Construction of plant transgenic vector pCAMBI1301a-PeVQ28 of Phyllostachys pubescens PeVQ28 gene

The PeVQ28 fragment, which was a small fragment ligated to pEASY T1simple Cloning Vector, was double digested with the Agrobacterium binary Vector pCAMBIA1301a using SmaI and Sal I enzymes in a 37 ℃ water bath for 3h, and the digested 20uL system is shown in Table 2:

TABLE 2

After double digestion, detection was performed by 1% agarose gel electrophoresis, and recovered by using an agarose gel DNA recovery kit. The large and small fragments are connected by T4DNA ligase, and are connected for 2-12 h at 25 ℃, and the connection system is shown in a table 3:

TABLE 3

Gently pumping 5-10 mu L of constructed vector pCAMBIA1301a-PeVQ28 plasmid into 100 mu L of EHA105 agrobacterium-infected cells, carrying out ice bath for 5min, quickly freezing for 1min by using liquid nitrogen, carrying out water bath at 37 ℃ for 5min, adding 200 mu L of YEP liquid culture medium, and carrying out culture at 28 ℃ and 220 for 4-5 h; 1.0X 10⁴g, centrifuging for 30s, removing the supernatant, adding 100 mu L YEP liquid culture medium, resuspending the cells, coating the cells on YEP solid plates containing 100 mu g/mL Kan and 50 mu g/mL Rif, and culturing for about 24-48 h at 28 ℃; picking a yellowish single colony growing on the plate, inoculating the yellowish single colony in YEP liquid culture solution containing 100 mu g/mL Kan and 50 mu g/mL Rif, and shaking the bacteria for 24-48 h; extracting plasmids when the bacterial liquid is turbid and shows orange yellow; verified by PCR and double digestion respectively.

Example 5

Plant transgenic expression vector pCAMBIA1301a-PeVQ28 is transferred into wild type arabidopsis

1. Planting of arabidopsis thaliana:

(1) rinsing the plump wild type Arabidopsis seeds with 12% King bleached water for 8min on a super clean bench, and repeatedly rinsing with sterilized distilled water for 7-8 times, each time for 3 min.

(2) The sterilized seeds were aspirated by a 1mL pipette and evenly blown on 1/2MS solid medium, vernalized in a 4 ℃ freezer for 3 days, and then transferred to a greenhouse for cultivation.

(3) One week later, arabidopsis on the plates were transplanted into small pots filled with vermiculite and black soil (3:1), 4 per pot. The plastic wrap was then covered for three days. Watering once in 3 days during the growth period to keep the soil in the flowerpot moist.

2. Preparation of agrobacterium infection liquid:

taking 50 mu L of the constructed over-expression vector agrobacterium liquid, adding the over-expression vector agrobacterium liquid into 10mLYEP liquid culture, simultaneously adding corresponding Kan and Rif, culturing for two days at 28 ℃, 230r/min, OD₆₀₀＝08-1.0。4℃，4.5×10³r/min, centrifuging for 6min, and discarding the supernatant. The pellet was resuspended in transformation buffer medium and ready for use.

The main stem which is bolting is cut off by scissors, the apical dominance is broken, and the main stem can have more lateral branches. When 1/3 flower buds are on the plant, a prepared bacterial liquid is absorbed by a rubber dropper, the flower buds are dipped, and existing fruit pods are cut by scissors, so that the false positive of seeds is reduced. Culturing in dark for three days. After about 5 days, the second infection was carried out by the same method. The final harvested seed was T0 generation.

Example 6

Transgenic arabidopsis screening and identification of phyllostachys pubescens PeVQ28 gene

1. Screening of hygromycin culture medium:

arabidopsis T0 seeds were evenly plated on hygromycin MS medium, as described above. After vernalization at 4 ℃ for 3 days, the cells were transferred to a greenhouse for culture. And observing the germination condition for about one week, wherein if the germination condition is normal, the plant is probably a transgenic positive plant.

PCR molecular validation

Firstly, extracting PeVQ28 transgenic Arabidopsis DNA according to a CTAB method,

(1) dissolving CTAB extraction buffer solution in a water bath (60 ℃);

(2) the leaves were placed in a mortar, poured into liquid nitrogen, and ground to a powder. Loading into 2mL tubes of the LDof, adding 1mL CTAB extraction buffer solution into each tube, shaking in vortex for 30s, and standing on ice for 3-5min for full lysis;

(3) placing the Dof tube in a water bath (65 ℃) and preserving heat for 1h to ensure that cells are completely broken;

(4) to the Dof tube was added 1mL phenol: chloroform: isoamyl alcohol (25: 24: 1), mixed well, 1.2X 10⁴g, centrifuging for 15min, taking the upper aqueous phase to a new Dof tube, adding equal volume of chloroform isoamylol, and fullyMixing, and 1.2 × 10⁴g, centrifuging for 15 minutes;

(5) sucking 400mL of supernatant into a new Dof tube, adding 400mL of isopropanol, mixing, standing in a refrigerator at-20 deg.C for half an hour (1.2 × 10)⁴g, centrifuging for 15min, and removing supernatant;

(6) adding 1mL of ethanol (70%), centrifuging for 6min, and removing the supernatant;

(7) repeating the step (6);

(8) adding 50. mu.L of sterilized pure water to dissolve the precipitate, and performing DNA purity, concentration and A₂₆₀/A₂₈₀And (4) detecting the value.

Wild type arabidopsis DNA was used as a control, primers PeVQ28F and PeVQ28R constructed using vectors, and PCR screening was performed on positive plant DNA of transgenic arabidopsis. PCR amplification procedure: 10min at 98 ℃; denaturation: 10s at 98 ℃; annealing: 5s at 62 ℃; extension: 30s at 72 ℃ for 28 cycles; total extension: 10min at 72 ℃. After the reaction is finished, taking the PCR product, carrying out 2% agarose gel electrophoresis, and detecting in a gel imaging system.

3. Phenotypic analysis of transgenic Arabidopsis thaliana of Phyllostachys pubescens PeVQ28 Gene

Planting the transgenic and wild arabidopsis homozygote seeds in a greenhouse, and photographing to observe the phenotype after two weeks; and simultaneously carrying out salt stress treatment, and photographing to observe the phenotypic change in the salt treatment process. And the survival rate of the arabidopsis thaliana of different strains after salt treatment is counted, and the physiological indexes before and after the dry salt treatment are counted (figure 4).

The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.

Sequence listing

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<120> Phyllostachys pubescens PeVQ28 protein and coding gene and application thereof

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gtgttggaag ttcttgcacc cttggttgca aataatgatg tgtatgtaag gggtcactgg 780

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gagcaccaat ttagttttga gcccctttca gtagacctgc caaagaatgc tctgattaag 1200

tgtttgcttc agtttactaa ttgcaaagga tgtgatcaaa caggtgtcaa tgactctatg 1260

cacaactgga aattaccttg atgggaataa ctctttttta atagcattct tctatgtgaa 1320

gtttttttta gtccggatgt gtgctgccat tagcaatatg cagaggtgta agtaagctgg 1380

cagaaaacga cgcttacata ggctaatgcc ctagctcact tgtttaataa gtattatatg 1440

ttcggctatc ttaaacaaga tccaatggtt ttcttctaat atgtcctcaa tttggaagaa 1500

ccatataata gtactgacct aagatgtact acagaaaaga cagatagata gcagtgctga 1560

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gccgtcgact taagatgcgt acatttcac 29

Claims

1. An application of a Phyllostachys Pubescens PeVQ28 gene in improving salt resistance of plants, wherein the nucleotide sequence of the Phyllostachys Pubescens PeVQ28 gene is shown as SEQ ID No. 2.

2. Use according to claim 1, characterized in that: the gene of claim 1 is transferred into a plant by an agrobacterium-mediated inflorescence dip method.