CN104120138A

CN104120138A - Arabidopsis AtPGK2 gene for enhancing salt tolerance of plants and application of arabidopsis AtPGK2 gene

Info

Publication number: CN104120138A
Application number: CN201410358406.7A
Authority: CN
Inventors: 刘栋; 李卫春; 马利霞; 程建峰
Original assignee: Jiangxi Agricultural University
Current assignee: Jiangxi Agricultural University
Priority date: 2014-07-26
Filing date: 2014-07-26
Publication date: 2014-10-29
Anticipated expiration: 2034-07-26
Also published as: CN104120138B

Abstract

The invention clones an arabidopsis 3-phosphoglycerate kinase gene AtPGK2 for enhancing salt tolerance of plants, and discloses an application of the arabidopsis 3-phosphoglycerate kinase gene AtPGK2. The nucleotide sequence of the gene is shown in SEQIDNO: 1, and the gene also comprises genes which have homologies of 90-100% with the SEQIDNO: 1 nucleotide sequence. Meanwhile, the invention also provides a construction and transgenosis method of a recombinant vector to apply the gene. The method can be used for cultivating new varieties of transgenic plants with stronger salt tolerance, and has wide application values.

Description

A kind of Arabidopis thaliana AtPGK2 gene and application thereof that strengthens plant salt endurance

Technical field

The invention belongs to plant genetic engineering field, is to utilize Protocols in Molecular Biology to obtain a kind of gene that can strengthen plant salt endurance specifically atPGK2and the application in transgenic plant.

Background technology

The soil salinization is all very serious in the whole world, and salt stress is one of the important abiotic stress of the global crop yield of impact (Parvaiz and Satyawati, 2008).China's saline soil area is large, distribution is wide, type is many, account for 10% of area, and annual salinization and Secondary Saline are all constantly increasing the weight of, the Sustainable development of agriculture production is on the hazard, have a strong impact on growing and the yield and qualities (Sun Jianchang etc., 2008) of cash crop of plant.Therefore the transgenic crop new variety that, cultivation salt tolerance strengthens are very urgent tasks.

Plant genetic engineering refers in vitro, utilizes toolenzyme to carry out to DNA molecular the DNA molecular that " montage " and " splicing " forms restructuring, by its transformed plant cells and make its expression.At present, the transgenic plant new variety of utilizing plant genetic engineering to cultivate salt tolerance enhancing have obtained certain progress.Much research shows, the salt-resistant related gene of plant is imported to the cell of self or other species and produces transfer-gen plant, can significantly strengthen the salt resistance ability of transgenic plant.As overexpression corn in Arabidopis thaliana zmCBL4gene can improve transfer-gen plant at germination period and Seedling Stage to the tolerance of salt stress (Wang et al., 2007); Chen etc. (2008) utilize constitutive promoter caMV 35Sdrive Arabidopis thaliana atNHX1gene is overexpression in buckwheat, and in transgenosis buckwheat plant, the content of main nutrient composition is not subject to the impact of high-salt stress; Liang etc. (1997) utilize caMV 35Spromoters driven spinach bADHgene is overexpression in tobacco plant, and the salt tolerance of transgene tobacco significantly improves.

Glycerol 3-phosphate acid kinase (the 3-Phosphoglycerate kinase of plant; PGK) as important enzyme catalysis 3-phoshoglyceric acid and 1; change between 3-diphosphoglyceric acid has critical function (Blake and Rice, 1981 in carbon fixation, glycolysis-and glyconeogenesis metabolism; Watson et al., 1982).At present, the gene of many coding PGKs is cloned (Watson and Littlechild, 1990) in plant, animal and microorganism.In plant, as pea (Pacold and Anderson, 1975), wheat (Longstaff et al., 1989), barley (McMorrow and Bradbeer, 1990), spinach (Kopke-Secundo et al., 1990) and the PGKs of tomato (Rao et al., 1995) etc. separated and clone successively, and the albumen majority of its coding is purified for enzymic activity research.Sequential analysis shows, the PGKs of higher plant is encoded by two cell nucleus gene, the aminoacid sequence of its proteins encoded has higher homology, wherein, the aminoacid sequence of a genes encoding at N-end by one section of transit peptides that is positioned chloroplast(id) approximately being formed by 70 amino acid, on the contrary, the aminoacid sequence of another genes encoding there is no transit peptide sequence at N-end.Therefore, infer that two PGKs that encoded by higher plant cell core are positioned respectively chloroplast(id) and tenuigenin (Longstaff et al., 1989; Kitayama and Togasaki, 1995).The above concentrates on the later stage in last century to the enzymic activity research majority of the separation of higher plant PGKs gene, clone, sequential analysis and its proteins encoded.Enter this century, although there are some researches show in the recent period under condition of salt stress, in plant materials, the content of PGK albumen significantly increases (Kosova et al., 2011; Wang et al., 2013), but for its physiological function particularly the function of involved in plant response salt stress it be unclear that.

Plant is for adapting to salt stress; to reduce to greatest extent the injury of salt stress to self; from the expression to final regulation and control stress response gene to the perception of salt stress signal, intracellular signal transmission and intracellular signal transduction; salt stress (Zhu, 2002 are resisted in the variation that makes plant produce a series of forms, physiology and chemistry; Jiang and Deyholos, 2006; Radi et al., 2013).In order to excavate plant salt tolerance gene and to cultivate the transgenic plant that salt tolerance strengthens, the present invention has cloned Arabidopis thaliana coding 3-phoshoglyceric acid kinase gene atPGK2, by this gene transformation model plant Arabidopis thaliana, find that transgenic arabidopsis plant obviously strengthens the tolerance of salt stress.

Summary of the invention

The present invention seeks to the resistant gene of salt of separated and clone plant, and the function of its participation salt stress response is identified, a kind of molecular biology model plant Arabidopis thaliana 3-phoshoglyceric acid kinase gene is provided atPGK2and the application in transgenic plant.

The invention provides a kind of Arabidopis thaliana 3-phoshoglyceric acid kinase gene new, 1437 bp atPGK2sequence, the nucleotide sequence of this gene is selected from:

(a) nucleotide sequence as shown in sequence table SEQ ID NO:1;

(b) in the nucleotide sequence limiting at (a), there is the homology nucleotide sequence between 90～100%.As the example of concrete application, the invention provides a kind of Arabidopis thaliana coding 3-phoshoglyceric acid kinase gene atPGK2cloning process, concrete operation step is as follows:

(1) total RNA the reverse transcription of extraction Arabidopis thaliana plant are cDNA;

(2) take cDNA as template, by PCR method, increase atPGK2the CDS sequence of gene;

(3) reclaim pcr amplification product.

The present invention provides Arabidopis thaliana 3-phoshoglyceric acid kinase gene simultaneously atPGK2the application of coercing at plant salt endurance utilizes clone's that is: atPGK2the CDS sequence of gene, build " caMV 35S- atPGK2" fusion gene, carry out Plant Transformation, thereby obtain constitutive expression in Arabidopis thaliana atPGK2the transgenic plant of goal gene.Goal gene in wherein said fusion gene can be any goal gene needing for fundamental research, transformation technology, flowers or the improvement of farm crop salt-tolerance character.

As the example of concrete application, the invention provides a kind of " caMV 35S- atPGK2" fusion gene structure and at transgenic arabidopsis, participate in the application of salt stress.Specific operation process is as follows:

(1) use ncoi/ bstEiI double digestion passes through pcr amplification atPGK2the CDS sequence fragment of gene;

(2) use ncoi/ bstEiI double digestion pCAMBIA 1301(is purchased from CAMBIA company) plasmid, reclaims large carrier segments;

(3) mix that the above-mentioned the first step obtains atPGK2the CDS sequence fragment of gene and the pCAMBIA that second step obtains 1301 carrier large fragments are carried out ligation under ligase enzyme catalysis, complete on pCAMBIA 1301 carriers " caMV 35S- atPGK2" structure of fusion gene.

Wherein designed atPGK2the pcr amplification primer of the CDS sequence of gene is as follows, and wherein upstream primer has been introduced ncoi restriction enzyme site, downstream primer has been introduced bstEiI restriction enzyme site:

Upstream primer: 5 '-CATG cCATGGcTTCCACCGCCGCAACTGCA-3 '

Downstream primer: 5 '-G gGTAACCtTAAACAGTGACTGGCGTTGCT-3 '.

In described application " caMV 35S- atPGK2" constitutive expression that is structured in transgenic arabidopsis of fusion gene expresses, operating process is as follows:

The concrete grammar of transformation of Arabidopsis thaliana, adopts the method (Clough and Bent, 1998) of agriculture bacillus mediated Floral dip, and the seed of acquisition is through 50 mg l ^-1hygromycin resistance screening, the normal resistant plant of growing turns earth culture supports, and utilizes 50 mg l ^-1totomycin carry out resistance screening and to transform atPGK2the transgenic arabidopsis strain of isozygotying of gene, then detects the salt tolerance of transgenic arabidopsis strain.

Positively effect of the present invention:

Test-results shows, by clone's coding 3-phoshoglyceric acid kinase gene atPGK2transformation mode plant Arabidopis thaliana, can significantly improve the salt tolerance of transfer-gen plant.Fusion gene construction of the present invention " caMV 35S- atPGK2" can be used as that genetic resources is used in agriculture production or seed selection transgenic plant to improve salt tolerance, be with a wide range of applications.

Accompanying drawing explanation

Fig. 1 be transform " caMV 35S- atPGK2" goal gene in the transgenic arabidopsis strain of fusion gene atPGK2expression analysis, * * P ﹤ 0.01.

Fig. 2 be transform " caMV 35S- atPGK2" Salt Tolerance Analysis of transgenic arabidopsis strain Seedling Stage of fusion gene.

Fig. 3 be transform " caMV 35S- atPGK2" the transgenic arabidopsis strain of fusion gene becomes the Salt Tolerance Analysis in seedling stage.*P﹤0.05。

Embodiment

embodiment 1:arabidopis thaliana coding 3-phoshoglyceric acid kinase gene atPGK2clone

(1) utilize different sulfuric acid nitrile guanidine-phenol method to extract total RNA of Arabidopis thaliana plant, the use of medicine, preparation and concrete operation step are with reference to the method for (2002) such as Huang Peitang.

(2) according to known Arabidopis thaliana atPGK2the CDS sequences Design special primer of gene is introduced in upstream primer ncoi restriction enzyme site is introduced in downstream primer bstEiI restriction enzyme site.

Upstream primer: 5 '-CATG cCATGGcTTCCACCGCCGCAACTGCA-3 ' (introduces ncoi restriction enzyme site)

Downstream primer: 5 '-G gGTAACCtTAAACAGTGACTGGCGTTGCT-3 ' (introduces bstEiI restriction enzyme site)

(3) get the template that 1 mg RNA does reverse transcription, with P ₂₈₅₃for primer utilizes the ThermoScript II ImProm-II of Promega ^tMcarry out reverse transcription, P ₂₈₅₃primer sequence, reverse transcription program and reverse transcription system are as follows.

P ₂₈₅₃primer sequence: 5'-GCGAATTCTTTTTTTTTTTTTTTTT-3'

Reverse transcription program:

72 ℃ 5 minutes; 25 ℃ 5 minutes; 42 ℃ 60 minutes; 80 ℃ 20 minutes; 4 ℃ of insulations.

Reverse transcription system:

Reagent	Add-on (μ l)
		DEPC water	5.0
P ₂₈₅₃Primer (10 μ mol l ^-1）	1.0
		5 * reaction buffer is (containing Mg ²⁺）	4.0
MgCl ₂（25 mmol l ^-1）	2.4
		dNTP（10 mmol l ^-1）	4.0
HPRI inhibitor	0.6
		ImProm-II ^TMThermoScript II (5 U μ l ^-1）	1.0
Template ribonucleic acid (50 ng μ l ^-1）	2.0

(4) take the cDNA of reverse transcription is template, utilizes above-mentioned primer to carry out pcr amplification, obtains atPGK2the CDS fragment of gene.

PCR response procedures:

94 ℃ 3 minutes;

94 ℃ 30 seconds, 56 ℃ 30 seconds, 72 ℃ 2 minutes, 30 circulations;

72 ℃ 10 minutes;

4 ℃ of insulations.

PCR reaction system:

Reagent	Add-on (μ l)
		Sterilizing distilled water	36.5
10 * reaction buffer is (containing Mg ²⁺）	5.0
		dNTP（10 mmol l ^-1）	4.0
Upstream primer (10 μ mol l ^-1）	1.0
		Downstream primer (10 μ mol l ^-1）	1.0
ExTaq archaeal dna polymerase (5 U μ l ^-1）	0.5
		Template DNA (50 ng μ l ^-1）	2.0

(5) by agarose gel electrophoresis, reclaim target DNA fragment, recovery method adopts the DNA sepharose of the precious biotech firm in Dalian to reclaim test kit, and concrete operation step is shown in catalogue.

(6) the PCR product after reclaiming is carried out to DNA sequencing, its nucleotide sequence is as shown in sequence table 1.

embodiment 2:utilize pCAMBIA1301 vector construction " caMV 35S- atPGK2" fusion gene

(1) from intestinal bacteria, extract carrier pCAMBIA 1301 plasmids (purchased from CAMBIA company), use ncoi/ bstEafter II double digestion, reclaim large carrier segments.

(2) embodiment 1 is reclaimed atPGK2the CDS fragment of gene is used ncoi/ bstEiI double digestion, reclaims (with embodiment 1) fragment after enzyme is cut by agarose gel electrophoresis.

(3) above-mentioned 2 fragments are spent the night in 16 ℃ of connections under ligase enzyme catalysis, complete on pCAMBIA 1301 carriers " caMV 35S- atPGK2" Fusion gene construction.

Linked system:

Reagent	Add-on (μ l)
		AtPGK2The CDS fragment of gene (50 ng μ l ^-1）	2.0
PCAMBIA1301 carrier large fragment (50 ng μ l ^-1）	3.0
		Solution I ligase enzyme	5.0

(4) with connecting mixture, transform bacillus coli DH 5 alpha competent cell, concrete grammar is as follows:

CaCl routinely ₂induction and method for transformation, prepare bacillus coli DH 5 alpha competent cell, with 10 μ l, connects product transformed competence colibacillus cell, is then evenly applied on the flat board that contains Amp, X-gal and IPTG, is inverted for 37 ℃ and cultivates 12 hours.

(5) take plasmid carries out PCR reaction as template, identify in plasmid " caMV 35S- atPGK2" fusion gene, the size of amplified fragments is 1678 bp.The primer is as follows:

Upstream primer: 5'-GCGATAAAGGAAAGGCCATCG-3'

Downstream primer: 5 '-GGGTAACCTTAAACAGTGACTGGCGTTGCT-3 '

(6) from positive colony, extract plasmid, by ordinary method, transform Agrobacterium GV3101, obtain through engineering approaches Agrobacterium, for Plant Transformation.

embodiment 3:the preparation of transgenic arabidopsis plant

(1) with embodiment 2, build " caMV 35S- atPGK2" fusion gene arabidopsis thaliana transformation, concrete method for transformation adopts the method (Clough and Bent, 1998) of agriculture bacillus mediated Floral dip, and the seed of acquisition is through 50 mg l ^-1hygromycin resistance screening, the normal plant that grows turns earth culture and supports.

(2) real-time quantitative RT-PCR of transfer-gen plant detects: the transgenic arabidopsis expansion of identifying through PCR in embodiment 2 is numerous, and through 50 mg l ^-1hygromycin resistance screening obtains T ₃the transgenic line that generation isozygotys, obtains respectively growth wild-type and the transgenic arabidopsis seedling of 1 week, and total RNA the reverse transcription of according to the method for embodiment 1, extracting plant tissue are mRNA, by following primer, response procedures and reaction system, carry out PCR reaction:

atPGK2the detection primer of gene:

Upstream primer: 5 '-CGTTGACTCTCGTTTCTCGGTCC-3 '

Downstream primer: 5 '-TCCAACACTCTTCTTCGCCATCG-3 '

tIP41-likethe detection primer of gene:

Upstream primer: 5 '-GTATGAAGATGAACTGGCTGACAAT-3 '

Downstream primer: 5 '-ATCAACTCTCAGCCAAAATCGCAAG-3 '

Response procedures:

95 ℃: 2 minutes; 1 circulation; 95 ℃: 10 seconds; 60 ℃: 30 seconds; 40 circulations.

Reaction system:

Composition	Add-on (m L）
		Sterilizing distilled water	4.0
2 x SYBR Premix	10.0
		cDNA（10 ng/mL）	2.0
Upstream primer (2 μ mol l ^-1）	2.0
		Downstream primer (2 μ mol l ^-1）	2.0

Real-time quantitative PCR result as shown in Figure 1, atPGK2gene significantly raises at the expression amount of two transgenic lines, and comparing rising degree with wild-type increases by 7～8 times.

embodiment 4:cross expression atPGK2the salt tolerance of the transgenic arabidopsis strain of gene detects

(1) Seedling Stage salt tolerance detects:

The transgenic line of identifying by real-time quantitative RT-PCR in embodiment 3 is carried out to salt tolerance detection at Seedling Stage.Concrete grammar: by wild-type and the expression of crossing of isozygotying atPGK2gene T ₃the transgenic arabidopsis seed in generation is layered on 1/2MS substratum germination and growth 4 days (growth conditions: intensity of illumination: 90 μ E m ^-2s ^-1, the photoperiod: 16 h illumination/8 h are dark), then seedling is transferred on the 1/2MS substratum that contains 150 mM NaCl and grows 15 days, every the survival rate (sum of the seedling number/seedling of survival rate=vegetative point necrosis) of 3 days statistics seedling.Test-results as shown in Figure 2, is crossed and is expressed atPGK2the transgenic arabidopsis strain of gene increases significantly compared with wild-type Arabidopis thaliana in the salt tolerance of Seedling Stage.

(2) become Seedling Salt-tolerance to detect:

The transgenic line of identifying by real-time quantitative RT-PCR in embodiment 3 is being become to carry out salt tolerance detection seedling stage.Concrete grammar: by wild-type and the expression of crossing of isozygotying atPGK2gene T ₃the transgenic arabidopsis seed in generation is sowed in soil, cultivates the 28 days (growth conditionss: intensity of illumination: 90 μ E m of growing under normal illumination condition ^-2s ^-1, the photoperiod: 16h illumination/8h is dark), then every the pouring of the NaCl solution with 300 mM in 3 days, within 15 days, add up afterwards the survival rate (sum of the number of seedling/seedling of survival rate=vegetative point necrosis) of seedling.Test-results as shown in Figure 3, is crossed and is expressed atPGK2the transgenic arabidopsis strain of gene is becoming the salt tolerance in seedling stage to increase significantly compared with wild-type Arabidopis thaliana.

The above embodiments result shows and confirms, Arabidopis thaliana provided by the present invention atPGK2the salt tolerance of gene and plant is proportionate, and in plant, expressing this gene can significantly improve transgenic plant in Seedling Stage and the salt tolerance that becomes seedling stage excessively.Therefore, Fusion gene construction of the present invention " caMV 35S- atPGK2" can be used as that genetic resources is used in agriculture production or seed selection transgenic plant to improve salt tolerance, be with a wide range of applications.

Sequence table

SEQUENCE LISTING

<110> Agricultural University Of Jiangxi

<120> Arabidopis thaliana AtPGK2 gene and application thereof that strengthens plant salt endurance

<130> 2014

<160> 9

<170> PatentIn version 3.3

<210> 1

<211> 1437

<212> DNA

<213> Arabidopis thaliana (Arabidopsis thaliana)

<400> 1

atggcttcca ccgccgcaac tgcagctctt tcgatcatca aatccaccgg tggcgccgcc 60

gttacacgct cctcccgcgc ctcctttgga cacattccct ccacatctgt ctccgcacgt 120

cgccttggct tctccgccgt cgttgactct cgtttctcgg tccacgtggc gtccaaggtc 180

cactccgtgc gcggaaaggg cgccagggga gtgattacga tggcgaagaa gagtgttgga 240

gatctgaatt ctgtagattt gaaggggaag aaggtgtttg ttagagctga tctcaatgta 300

cctctcgatg acaaccagaa tatcactgac gacactagaa tcagagccgc cattcccacc 360

atcaagtttt tgattgagaa tggtgctaag gtcatcctct ccactcattt gggaaggcca 420

aagggtgtca ctccaaagtt cagcttggct cctcttgttc ccagattatc agagctcctt 480

ggtattgagg tcgtgaaagc tgatgattgt attggtccag aagtggaaac cttggtggct 540

tcccttcctg aaggtggagt tttgcttctt gagaacgtga ggttttacaa ggaggaagag 600

aagaacgaac ctgattttgc taagaagctt gcttctctag ctgaccttta tgtcaacgat 660

gcgttcggga ctgctcacag agcccatgct tcaacagagg gagtcactaa gttcttgaag 720

ccatcagttg ctggtttcct tttgcaaaag gagttggact accttgttgg tgcggtttca 780

aacccaaaga gaccatttgc tgccattgtg ggaggttcca aggtctcatc taagattgga 840

gttatcgaat cgcttcttga gaaatgtgac atccttctgc ttggtggtgg aatgatcttt 900

acattctaca aggcgcaagg tctttccgtt ggctcctccc ttgttgaaga agacaagctt 960

gaattggcta caacactcct tgccaaggct aaggccagag gagtctctct gttgttacca 1020

acagatgttg tgattgctga caagttcgct cctgatgcca acagcaagat tgtgccagca 1080

tcagccattc ctgatgggtg gatgggattg gacatcggtc cagactcggt gaaaacattc 1140

aacgaagctc tggataccac gcagacagtc atttggaatg gaccaatggg agttttcgag 1200

tttgaaaagt ttgcaaaagg aactgaggcg gtagcgaata aactagcaga gctaagcaaa 1260

aagggagtga caacgataat aggaggagga gactcggtgg cagcagtgga gaaagtggga 1320

gtagcaggag tcatgagtca catctccaca ggtggtggtg ccagtttgga gctcttggaa 1380

ggcaaagtgc ttcccggtgt cgtcgctctt gatgaagcaa cgccagtcac tgtttaa 1437

<210> 2

<211> 30

<212> DNA

<213> artificial sequence

<400> 2

catgccatgg cttccaccgc cgcaactgca 30

<210> 3

<211> 30

<212> DNA

<213> artificial sequence

<400> 3

gggtaacctt aaacagtgac tggcgttgct 30

<210> 4

<211> 25

<212> DNA

<213> artificial sequence

<400> 4

gcgaattctt tttttttttt ttttt 25

<210> 5

<211> 21

<212> DNA

<213> artificial sequence

<400> 5

gcgataaagg aaaggccatc g 21

<210> 6

<211> 23

<212> DNA

<213> artificial sequence

<400> 6

cgttgactct cgtttctcgg tcc 23

<210> 7

<211> 23

<212> DNA

<213> artificial sequence

<400> 7

tccaacactc ttcttcgcca tcg 23

<210> 8

<211> 25

<212> DNA

<213> artificial sequence

<400> 8

gtatgaagat gaactggctg acaat 25

<210> 9

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atcaactctc agccaaaatc gcaag 25

Claims

1. an Arabidopis thaliana that strengthens plant salt endurance atPGK2gene, is characterized in that, is one of following nucleotide sequences:

(a) nucleotide sequence as shown in sequence table SEQ ID NO:1;

(b) in the nucleotide sequence limiting at (a), there is the homology nucleotide sequence between 90～100%.

2. strengthen as claimed in claim 1 the Arabidopis thaliana of plant salt endurance atPGK2the cloning process of gene, concrete operation step is as follows:

(3) reclaim pcr amplification product.

3. the Arabidopis thaliana of enhancing plant salt endurance as claimed in claim 1 atPGK2the application of gene, it is to utilize clone's atPGK2the CDS sequence of gene, build " caMV 35S- atPGK2" fusion gene, carry out Plant Transformation, thereby obtain constitutive expression in Arabidopis thaliana atPGK2the transgenic plant of goal gene, the goal gene in wherein said fusion gene can be any goal gene needing for fundamental research, transformation technology, flowers or the improvement of farm crop salt-tolerance character; " caMV 35S- atPGK2" fusion gene structure and at transgenic arabidopsis, participate in the application of salt stress, specific operation process is as follows:

(3) mix that the above-mentioned the first step obtains atPGK2the CDS sequence fragment of gene and the pCAMBIA that second step obtains 1301 carrier large fragments are carried out ligation under ligase enzyme catalysis, complete on pCAMBIA 1301 carriers " caMV 35S- atPGK2" structure of fusion gene;

Upstream primer: 5 '-CATG cCATGGcTTCCACCGCCGCAACTGCA-3 '

Downstream primer: 5 '-G gGTAACCtTAAACAGTGACTGGCGTTGCT-3 ';

In described application " caMV 35S- atPGK2" fusion gene be structured in constitutive expression in transgenic arabidopsis, operating process is as follows: the concrete grammar of transformation of Arabidopsis thaliana, adopt the method for agriculture bacillus mediated Floral dip, the seed of acquisition is through 50 mg l ^-1hygromycin resistance screening, the normal resistant plant of growing turns earth culture supports, and utilizes 50 mg l ^-1totomycin carry out resistance screening and to transform atPGK2the transgenic arabidopsis strain of isozygotying of gene, then detects the salt tolerance of transgenic arabidopsis strain;

Described plant is wheat, barley, paddy rice, corn, soybean, peanut, cotton, potato, rape, tomato, cucumber, clover or Arabidopis thaliana.