CN103709238B - Application of poplar salt tolerant gene PtoeIF5A1 - Google Patents

Application of poplar salt tolerant gene PtoeIF5A1 Download PDF

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CN103709238B
CN103709238B CN201310700593.8A CN201310700593A CN103709238B CN 103709238 B CN103709238 B CN 103709238B CN 201310700593 A CN201310700593 A CN 201310700593A CN 103709238 B CN103709238 B CN 103709238B
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ptoeif5a1
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arabidopis thaliana
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CN103709238A (en
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张杰伟
魏建华
王宏芝
李瑞芬
张中保
陈亚娟
丁莉萍
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Beijing Academy of Agriculture and Forestry Sciences
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    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance

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Abstract

The invention discloses an application of a poplar salt tolerant gene PtoeIF5A1. The invention discloses an application of any one of the following substances in improvement of plant salt tolerance: (1) a protein represented in SEQ ID No. 2; (2) a coding gene of the protein represented in SEQ ID No. 2; and (3) a recombinant vector, an expression cassette, a transgenic cell line or a recombinant bacterium containing the (2). The PtoeIF5A1 gene is high in salt tolerance, and has high application value in improving salt tolerance of forests or crops through a genetic engineering means.

Description

The application of willow resistant gene of salt PtoeIF5A1
Technical field
The present invention relates to the application of a kind of willow resistant gene of salt PtoeIF5A1.
Background technology
EIF5A is the class protein extensively existed in eukaryote, and research shows, eIF5A is the protein uniquely containing carboxylic putrescine Methionin (hypusine) residue.Evolutionary analysis shows, and eIF5A is high conservative between each species.Experiment shows, eIF5A take part in the multinomial physiological activity of cell, as cell proliferation, protein translation, cell aging and apoptosis etc.In yeast body, when eIF5A function is suppressed or lacks, Partial Protein synthesis reduces.When the activity of eIF5A is suppressed, the delayed growth of cell or stagnation.
The biological function that in plant, eIF5A is definite is also indefinite.The research such as Thompson finds to there is the different eIF5A encoding gene of 3 expression characteristics in Arabidopis thaliana, AteIF5A1, AteIF5A2 and AteIF5A3.WesternBlotting analyzes and shows, AteIF5A1 only expresses in aging tissues, and AteIF5A2 is high level expression in the tissue be mechanically damaged, and AteIF5A3 high level expression in the seed of the very active imbibition of cell fission.
Summary of the invention
The object of this invention is to provide the application of a kind of willow resistant gene of salt PtoeIF5A1.
The invention provides following arbitrary material and improve the application in plant salt endurance:
(1) protein shown in SEQ ID No.2;
(2) encoding gene of protein shown in SEQ ID No.2;
(3) recombinant vectors containing (2), expression cassette, transgenic cell line or recombinant bacterium.
In above-mentioned application, shown in described SEQ ID No.2, the encoding gene of protein is as shown in SEQ ID No.1.
In above-mentioned arbitrary described application, described salt tolerance is the character of resistance to NaCl.
In above-mentioned arbitrary described application, described raising plant salt endurance refers to that described plant is good containing the growth conditions under NaCl condition; And/or, high containing the seed germination rate under NaCl condition;
Described growth conditions refers to that well the blade of described plant is comparatively large, color is greener;
In above-mentioned arbitrary described application, described plant is Arabidopis thaliana.
The method preparing the transgenic plant that salt tolerance improves also belongs to a protection scope of the present invention, comprises the steps: the encoding gene of protein shown in SEQ ID No.2 to import to set out in plant, obtains transgenic plant; Compared with the plant that sets out, the salt tolerance of transgenic plant strengthens.
In aforesaid method, described encoding gene is imported by recombinant expression vector, and described recombinant expression vector is that the multiple clone site described encoding gene being inserted the carrier pBI121 that sets out obtains.
In above-mentioned arbitrary described method, the salt tolerance of described transgenic plant strengthens and refers to that described transgenic plant are being better than the plant that sets out containing the growth conditions under NaCl condition containing the growth conditions under NaCl condition; And/or the seed of transgenic plant is containing the germination rate under NaCl condition containing the germination rate under NaCl condition higher than the plant that sets out.
The concentration of described NaCl is 150mM-175mM.
In above-mentioned arbitrary described method, described plant is Arabidopis thaliana.
In above-mentioned arbitrary described method, described salt tolerance is the character of resistance to NaCl.
Present invention demonstrates that PtoeIF5A1 gene has the ability tolerating high salt, this gene has using value in use genetic engineering means raising forest or crop tolerance to salt.
Accompanying drawing explanation
Fig. 1 be Western-Blotting detect PtoeIF5A1 albumen at transgenic arabidopsis T0 for plant and contrast Arabidopis thaliana T0 for the expression in plant.
Fig. 2 is the expression that real-time fluorescence quantitative PCR detects PtoeIF5A1 gene.
Fig. 3 is that isozygoty in the T2 generation Arabidopis thaliana that turns PtoeIF5A1 gene and T2 generation the isozygoty salt tolerance that turns empty carrier pBI121 Arabidopis thaliana detects and the Western-Blotting of PtoeIF5A1 albumen detects.
Fig. 4 is seed germination experiment.
Embodiment
The experimental technique used in following embodiment if no special instructions, is ordinary method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
Turriform Cortex Populi Tomentosae CV-BJHR01(Populus tomentosa Carr.cv ' BJHR01 ') document " Zhang Lijiao; Zhang Jiewei; Chen Yajuan; etc. cloning and expression analysis [J] the .Journal of Agricultural Biotechnology of Cortex Populi Tomentosae Eukaryotic initiation factor 5A gene (PtoeIF5A4); 2013; 21 (8): 949-956. " in be disclosed, the public can obtain from Beijing City Agriculture and Forestry Institute.
Arabidopis thaliana (Arabidopsis thaliana, Columbia ecotype) be disclosed in document " Initiative A G.Analysis of the genome sequence of the flowering plant Arabidopsis thaliana [J] .Nature; 2000; 408 (6814): 796-815. ", the public can obtain from Beijing City Agriculture and Forestry Institute.
Cloning vector pGEM-T easy is purchased from Promega company, and catalog number is A1360.
PBSK-Ω-Flag carrier (modified pBluescript vector) is disclosed in document " Su T; Xu J; Li Y; et al.Glutathione-indole-3-acetonitrile is required for camalexinbiosynthesis in Arabidopsis thaliana [J] .The Plant Cell; 2011; 23 (1): 364-380. ", and the public can obtain from Beijing City Agriculture and Forestry Institute.
Plant expression vector pBI121 is disclosed in document " Su T; Xu J; Li Y; et al.Glutathione-indole-3-acetonitrile is required for camalexin biosynthesis inArabidopsis thaliana [J] .The Plant Cell; 2011; 23 (1): 364-380. ", and the public can obtain from Beijing City Agriculture and Forestry Institute.
Agrobacterium GV3101 is disclosed in document " Li J F; Li L; Sheen J.Protocol:a rapid and economicalprocedure for purification of plasmid or plant DNA with diverse applicationsin plant biology [J] .Plant Methods; 2010; 6 (1): 1-8. ", and the public can obtain from Beijing City Agriculture and Forestry Institute.
0.5%TBST solution: this solution is made up of solvent and solute, solvent is Tris-HCl damping fluid, solute is NaCl and Tween20; The concentration of Tris-HCl damping fluid is 50mM, the NaCl concentration in 0.5%TBST solution be 150mM, the Tween20 volumn concentration in 0.5%TBST solution is 0.05%, pH value of solution=7.5.
Primary antibodie: mouse-anti Flag M2 monoclonal antibody available from Sigma, is diluted 10000 times with 0.5%TBST solution during use.
Two resist: the sheep anti-mouse igg of horseradish peroxidase, purchased from Beijing Bioisystech Co., Ltd of Zhong Shan Golden Bridge, is diluted 10000 times with 0.5%TBST solution during use.
The structure of embodiment 1, pBSK-Ω-Flag-PtoeIF5A1 carrier
One, choose the blade that growth time is the turriform Cortex Populi Tomentosae CV-BJHR01 seedling of 3 months, extract total serum IgE after liquid nitrogen flash freezer and reverse transcription becomes cDNA.
Two, analyzed by comospore poplar genomic library (http: ∥ genome.jgi-psf.org/Poptr11/Poptr11.home.htm1), obtain the complete sequence of the Cortex Populi Tomentosae PtreIF5A1 gene of prediction, at its non-coding region design primer, the cDNA(of turriform Cortex Populi Tomentosae CV-BJHR01 obtained with step one is hereinafter referred to as Cortex Populi Tomentosae cDNA) carry out nested PCR amplification for template.
Nest-type PRC system is as follows:
First round PCR:
System: the PhusionDNA polysaccharase 0.2 μ L of upstream and downstream primer each 1 μ L, the 2.0U/L of Cortex Populi Tomentosae cDNA2.0 μ L, 10 μm of ol/L, the dNTPs mixed solution 1.6 μ L of 2.5mmol/L, 5 × HF Buffer4.0 μ L, ddH 2o is 10.2 μ L.
Upstream primer: 5 '-CCTCTCTGGTACTCTCTGTG-3 ';
Downstream primer: 5 '-GCAGAACGTTACTTAAATGGGC-3 '.
PCR condition: 98 DEG C of 30s; 98 DEG C of 10s, 55 DEG C of 30s, 72 DEG C of 1min, 30 circulations; 72 DEG C of total elongation 10min.
Second takes turns PCR:
System: the Phusion archaeal dna polymerase 0.2 μ L of upstream and downstream primer each 1 μ L, the 2.0U/L of first round pcr amplification product 2.0 μ L, 10 μm of ol/L, the dNTPs mixed solution 1.6 μ L of 2.5mmol/L, 5 × HF Buffer4.0 μ L, ddH 2o is 10.2 μ L.
Upstream primer: 5 '-CTCTTGATCAATCGCCGCC-3 ';
Downstream primer: 5 '-CTACAACTATCAGCAGTCAACC-3 '.
PCR condition: 98 DEG C of 30s; 98 DEG C of 10s, 55 DEG C of 30s, 72 DEG C of 1min, 72 DEG C of total elongation 10min, 30 circulations.
Three, second take turns after PCR terminates, reclaim the fragment of 500bp and be connected on pGEM-T easy carrier, obtaining recombinant vectors, being checked order by recombinant vectors, result is correct, by this recombinant plasmid called after pGEM-T easy-PtoeIF5A1.
Four, design the coding region of primer clone PtoeIF5A1, introduce NdeI restriction enzyme site at 5 ' end of upstream primer.
Primer sequence is as follows:
Upstream primer PtoeIF5A1F:5 '- cATATGtCTGACGAGGAGCAT-3 '
Downstream primer PtoeIF5A1R:5 '-TTACTTGGGGCCAATGTCC-3 '
(sequence shown in underscore is that enzyme cuts recognition site)
Five, with pGEM-T easy-PtoeIF5A1 plasmid for template, carry out pcr amplification with PtoeIF5A1F and PtoeIF5A1R for primer, obtain PtoeIF5A1 gene, gene fragment is connected with pGEM-T easy carrier and obtains recombinant plasmid.Checked order by recombinant plasmid, order-checking is correct.
The nucleotide sequence of PtoeIF5A1 gene is as shown in SEQ ID No.1.The aminoacid sequence of PtoeIF5A1 albumen is as shown in SEQ ID No.2.
Six, with the site that NdeI and SpeI(SpeI site is on pGEM-T easy carrier) recombinant vectors that double digestion step 5 obtains, obtain PtoeIF5A1 gene fragment; With NdeI and SpeI double digestion pBSK-Ω-Flag carrier, obtain carrier large fragment; Gene fragment is connected with carrier large fragment, obtains pBSK-Ω-Flag-PtoeIF5A1 recombinant plasmid.
Seven, XhoI and SpeI double digestion pBSK-Ω-Flag-PtoeIF5A1, obtains Flag-Ω-PtoeIF5A1 gene fragment; XhoI and SpeI double digestion pBI121, obtains carrier large fragment; Gene fragment be connected with carrier large fragment, obtain pBI121-Ω-Flag-PtoeIF5A1 recombinant plasmid, send order-checking by plasmid, order-checking is correct.
Eight, by pBI121-Ω-Flag-PtoeIF5A1 transformation Agrobacterium GV3101, obtain recombinant bacterium, by recombinant bacterium upgrading grain, send order-checking, result proves that recombinant bacterium is correct.Simultaneously by empty carrier pBI121 transformation Agrobacterium GV3101, obtain contrasting recombinant bacterium.
Embodiment 2, bud infusion method arabidopsis thaliana transformation
Arabidopis thaliana transgenic method adopts the bud infusion method of the people such as Clough, and detailed process is as follows:
One, the recombinant bacterium that obtains of picking embodiment 1 and contrast recombinant bacterium list bacterium colony access (containing Kan50 μ g/ml, Gent25 μ g/ml) in 5ml LB respectively, cultivate one day for 28 DEG C.
Two, being transferred to 200ml respectively containing in same antibiotic LB with volume ratio 1:100 by cultivating the bacterium liquid obtained, continuing to be cultured to OD 600the centrifugal 10min of=1.0,5000g room temperature, to be resuspended in 200ml1/2MS substratum in (containing 50g/l sucrose, 200 μ l/l Silwet L-77) respectively by bacterial sediment, obtain recombinant bacterium and contrast recombinant bacterium suspension.
Three, will treat that genetically modified Arabidopis thaliana is inverted, 5min is soaked in the recombinant bacterium that the inflorescence of more than lotus throne leaf is obtained in step 2 or contrast recombinant bacterium suspension, period weak vibrations 2-3 time, obtain respectively turning the Arabidopis thaliana (hereinafter referred to as transgenic arabidopsis) of PtoeIF5A1 gene and turning the Arabidopis thaliana (hereinafter referred to as contrast Arabidopis thaliana) of empty carrier pBI121.
Four, transgenic arabidopsis and contrast Arabidopsis plant are placed horizontally in 22 DEG C of environment, basin alms bowl is sealed with lighttight plastics bag, taken out after about 16h, vertically cultivate until the T0 of results transgenic arabidopsis and contrast Arabidopis thaliana is for seed, seed is for subsequent use after drying at room temperature.
The screening of embodiment 3, transgenic arabidopsis
One, with volumn concentration be 70% aqueous ethanolic solution by transgenic arabidopsis and contrast Arabidopis thaliana T0 for seed treatment 1min, sterilizing washing 2min, then with clorox process 13min, then with aseptic washing 5 times, each 2min, is placed in 4 DEG C of vernalization 2-3 days by the seed after process.
Two, the seed after vernalization is evenly laid on (10g/l sucrose, 50 μ g/ml Kan, 8g/l agar, pH=5.7) in 1/2MS screening culture medium, move into 22 DEG C of incubator (16hr illumination, 8hr is dark, and intensity of illumination is 80-100Lux) sprout.
Three, the blade edge of transgenic arabidopsis seedling after about 6 days, root are longer, although and contrast Arabidopis thaliana and also can sprout in 1/2MS screening culture medium, seedling is yellow compared with contemporaneously transgenic seedlings blade, root is very short.Respectively the seedling of transgenic arabidopsis and contrast Arabidopis thaliana is moved on new 1/2MS agar plate (10g/l sucrose, 50 μ g/ml Kan, 8g/l agar, pH=5.7), continue to be cultured to appearance 4 leaves, moved in soil and grow.
Four, extract transgenic arabidopsis T0 and carry out Western-Blotting detection for plant and contrast Arabidopis thaliana T0 for the total protein of plant, primary antibodie is mouse-anti Flag M2 monoclonal antibody, this antibody can be combined with the fusion rotein of Flag-tag and PtoeIF5A1, two resist the sheep anti-mouse igg for horseradish peroxidase, and result as shown in Figure 1.
In Fig. 1, Vector representative contrast Arabidopis thaliana T0 is for plant; 1,2,3 three strain transgenic arabidopsis T0 are represented for plant.
Fig. 1 shows, transgenic arabidopsis T0 has the expression of the fusion rotein of Flag-tag and PtoeIF5A1 for plant, thus has the appearance of 19kD object band, and contrasts Arabidopis thaliana T0 and do not have object band for plant.
Select transgenic arabidopsis and contrast Arabidopis thaliana T0 and obtain T1 for seed for plant results.
Five, will contrast Arabidopis thaliana and transgenic arabidopsis T1 continues kind in 1/2MS screening culture medium (10g/l sucrose, 50 μ g/ml Kan, 8g/l agar, pH=5.7) for seed, select the resistance of Progeny plants and the non-resistance colony being separated (3:1) by Mendelian's rule, then identify that transgenic arabidopsis T1 is for plant with Western-Blotting.
Results are accredited as positive contrast Arabidopis thaliana and transgenic arabidopsis T1 for seed, transgenic arabidopsis T2 is obtained for plant by resistance and Protein Detection, results transgenic arabidopsis T2 is for seed, if T2 for seed in 1/2MS screening culture medium (10g/l sucrose, 50 μ g/ml Kan, 8g/l agar, pH=5.7) all there is resistance, then corresponding T2 is homozygote for seed; Contrast Arabidopis thaliana T2 is obtained for plant equally by Resistance detecting, results contrast Arabidopis thaliana T2 is for seed, if T2 for seed in 1/2MS screening culture medium (10g/l sucrose, 50 μ g/mlKan, 8g/l agar, pH=5.7) all there is resistance, then corresponding T2 is homozygote for seed, finally obtains the isozygoty Arabidopis thaliana strain that turns empty carrier pBI121 and T2 generation in T2 generation isozygoty and turn the Arabidopis thaliana strain (PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3) of PtoeIF5A1 gene by this method.
The detection of the real-time fluorescence quantitative PCR of embodiment 4, transfer-gen plant
One, in Arabidopis thaliana strain PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 that the T2 that Example 3 obtains turns PtoeIF5A1 gene for isozygotying and T2 generation, isozygoty and turn the Arabidopis thaliana strain of empty carrier pBI121, extract total serum IgE respectively, reverse transcription obtains cDNA.
Two, respectively with the cDNA of four plants for template, with primers F 1 and R1, real-time fluorescence quantitative PCR amplification is carried out to the cDNA of gene PtoeIF5A1, with primer PtoACT9-F and PtoACT9-R, real-time fluorescence quantitative PCR amplification is carried out to reference gene PtoACT9 simultaneously.3 repetitions are established in a parallel test.Utilize 2 -Δ Δ CTcalculate the relative expression quantity of PtoeIF5A1 gene in each strain.
Wherein Δ Δ CT=(C t.Target-C t.Actin) Time x-(C t.Target-C t.Actin) Time0
Time x represents random time point, and Time0 represents that the target gene of 1 times amount after PtoACT9 corrects is expressed.
Target represents PtoeIF5A1 gene, and Actin represents reference gene PtoACT9.
Real-time fluorescence quantitative PCR primer is as follows:
PtoeIF5A1 gene test primer:
F1：5’-TGTCACTTCGTTGGGATTGA-3’；
R1：5’-CAAGATCTTTCCCCTCACCA-3’；
Reference gene PtoACT9 detects primer:
PtoACT9-F：5’-TTGCTGACCGTATGAGCAAG-3’；
PtoACT9-R：5’-AATCCACATCTGCTGGAAGG-3’。
As shown in Figure 2, in Fig. 2, Vector represents T2 generation and isozygotys and turn the Arabidopis thaliana strain of empty carrier pBI121 result; 5-OE1,5-OE2 and 5-OE3 represent three T2 respectively and turn Arabidopis thaliana strain PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 of PtoeIF5A1 gene for isozygotying.
Fig. 2 shows, turn the Arabidopis thaliana not expressing gene PtoeIF5A1 of empty carrier pBI121, and the expression amount turning PtoeIF5A1 in the Arabidopsis plant of PtoeIF5A1 gene is all very high.
Embodiment 5, the salt tolerance of PtoeIF5A1 gene plant of turning
One, the seed of these three transgenic lines of Arabidopis thaliana strain PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 turning PtoeIF5A1 gene and the T2 generation seed turning empty carrier pBI121 Arabidopis thaliana strain (control group) that isozygotys of being isozygotied in 4 DEG C of vernalization T2 of 2 days generation is placed on the 1/2MS flat board containing 0mM NaCl, at 22 DEG C, cultivate under 16hr illumination/8hr dark condition, after 12 days, three T2 generation the isozygoty growth conditions of seedling of the Arabidopis thaliana strain turning PtoeIF5A1 gene and the seedling of control group plant obviously do not distinguish.
Two, the seed of these three transgenic lines of Arabidopis thaliana strain PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 turning PtoeIF5A1 gene and the T2 generation seed turning empty carrier pBI121 Arabidopis thaliana strain (control group) that isozygotys of being isozygotied in 4 DEG C of vernalization T2 of 2 days generation is placed on the 1/2MS flat board containing 150mM and 175mM NaCl, 22 DEG C, cultivate under 16hr illumination/8hr dark condition, after 12 days, compared with control group, the seedling turning PtoeIF5A1 gene strain all shows good growth conditions, and its blade is comparatively large, color is greener.
As shown in Figure 3A, Fig. 3 B is the position view of each group of seedling to result.
In Fig. 3 B, Vector represents control group seedling, and 5-OE1,5-OE2 and 5-OE3 represent the seedling of T2 for these three transgenic lines of Arabidopis thaliana strain PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 turning PtoeIF5A1 gene that isozygoty respectively.
Three, extract T2 generation isozygoty these three transgenic lines of Arabidopis thaliana strain PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 of turning PtoeIF5A1 gene and the T2 generation total protein turning empty carrier pBI121 Arabidopis thaliana (control group) that isozygotys and carry out Western-Blotting detection, primary antibodie is mouse-anti Flag M2 monoclonal antibody, this antibody can be combined with the fusion rotein of Flag-tag and PtoeIF5A1, and two resist the sheep anti-mouse igg for horseradish peroxidase.
Result as shown in Figure 3 C.
In Fig. 3 C, Vector represents control group Arabidopis thaliana; 5-OE1,5-OE2 and 5-OE3 represent three T2 respectively and turn Arabidopis thaliana strain PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 of PtoeIF5A1 gene for isozygotying.
Fig. 3 C shows, these three willow PtoeIF5A1 gene overexpression strains of PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 all can detect PtoeIF5A1 albumen, and object band do not detected in control group.
Four, seed germination experiment
In 4 DEG C of vernalization T2 of 2 days generation, is isozygotied the seedling of these three transgenic lines of Arabidopis thaliana strain PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 turning PtoeIF5A1 gene and T2 generation isozygoty the seed that turns empty carrier pBI121 Arabidopis thaliana strain (control group) be placed in respectively containing 0,150 and 175mM NaCl 1/2MS flat board on, 22 DEG C, cultivate under 16hr illumination/8hr dark condition, the germination rate (seed shows money or valuables one carries unintentionally) adding up each strain is started, continuously statistics one week from the 1st day.
Result as shown in Figure 4.In Fig. 4, Vector represents control group; 5-OE1,5-OE2 and 5-OE3 represent three T2 respectively and turn Arabidopis thaliana strain PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 of PtoeIF5A1 gene for isozygotying.
Fig. 4 A shows, T2 generation the isozygoty germination rate under 0mM NaCl condition of Arabidopis thaliana strain PtoeIF5A1-OE1, PtoeIF5A1-OE2 and PtoeIF5A1-OE3 that turns PtoeIF5A1 gene and control group Arabidopis thaliana there is no notable difference.
Fig. 4 B shows, obviously comparatively control group Arabidopis thaliana was high from the 3rd day for the Arabidopis thaliana strain PtoeIF5A1-OE1 that T2 turns PtoeIF5A1 gene for isozygotying, the germination rate of PtoeIF5A1-OE2 and PtoeIF5A1-OE3 strain under 150mM NaCl condition, for its 2.3 to 2.6 times, be all about 1.3 times of control group Arabidopis thaliana from the 4th day to the 7th day.
Fig. 4 C shows, obviously comparatively control group Arabidopis thaliana was high from the 3rd day for the Arabidopis thaliana strain PtoeIF5A1-OE1 that T2 turns PtoeIF5A1 gene for isozygotying, the germination rate of PtoeIF5A1-OE2 and PtoeIF5A1-OE3 strain under 175mM NaCl condition, for its 1.7 to 2 times, be about its 1.3 to 1.8 times from the 4th day to the 7th day.
Result shows that the overexpression of willow PtoeIF5A1 gene can improve the high salt ability of tolerance of plant.

Claims (6)

1. following arbitrary material is improving the application in plant salt endurance:
(1) protein shown in SEQ ID No.2;
(2) encoding gene of protein shown in SEQ ID No.2;
(3) recombinant vectors containing (2), expression cassette, transgenic cell line or recombinant bacterium;
Described plant is Arabidopis thaliana.
2. application according to claim 1, is characterized in that: shown in described SEQ ID No.2, the encoding gene of protein is as shown in SEQ ID No.1.
3. application according to claim 1 and 2, is characterized in that: described salt tolerance is the character of resistance to NaCl.
4. prepare a method for the transgenic plant that salt tolerance improves, comprise the steps: the encoding gene of protein shown in SEQ ID No.2 to import to set out in plant, obtain transgenic plant; Compared with the plant that sets out, the salt tolerance of transgenic plant strengthens;
Described plant is Arabidopis thaliana.
5. method according to claim 4, is characterized in that: described encoding gene is imported by recombinant expression vector, and described recombinant expression vector is that the multiple clone site described encoding gene being inserted the carrier pBI121 that sets out obtains.
6. method according to claim 5, is characterized in that: described salt tolerance is the character of resistance to NaCl.
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