CN105734024A - Wolfberry glutamyl cysteine synthetase and encoding gene and application - Google Patents
Wolfberry glutamyl cysteine synthetase and encoding gene and application Download PDFInfo
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Abstract
The invention discloses wolfberry glutamyl cysteine synthetase and an encoding gene and application.The amino acid sequence of wolfberry glutamyl cysteine synthetase is as shown in SEQ ID NO.1.The encoding gene of the wolfberry glutamyl cysteine synthetase is transferred into a target plant, and a transgenic plant high in cadmium ion resistance and high in salt resistance is obtained.
Description
Technical field
The present invention relates in a kind of Fructus Lycii (LyCiumChinenseMiller) gamma glutamyl cysteine synthetase (LcGCS) and encoding gene thereof and application.
Background technology
The biotic such as animals and plants, in normal growth process, not only can meet with saline and alkaline, heavy metal, arid, the abiotic stress such as hot and cold, also can run into wound, insect pest, virus.Under these stress conditions, internal can produce accumulated active oxygen, cause, cellular content, cell membrane lipid equivalent damage.Plant evolution goes out some regulatory mechanism to tackle these adverse circumstances, for instance produce substantial amounts of reducing substances and antioxidase.Animals and plants by regulate glutathion (Glutathione, GSH) synthetase series such as γ-GCS (Glutamylcys-teineSynthetase) expression and increase the synthesis of reducing substances glutathion.
GSH is the tripeptides of glutamic acid, cysteine and glycine composition[DixonDPetal.,2005], it is divided into oxidized form (GSSG) and reduced form (GSH).Usually said glutathion is reduced form, and it is very important antioxidant in plant, improves the ability of plant resistant adverse circumstance.GSH synthesis is main to be realized through two step dependency ATP catalytic reactions.Under ATP function, glutamic acid, cysteine and glycine energy supply synthesize GSH under γ-GCS and GS (Glutathionesynthase) catalysis.When cysteine is sufficient, γ-GCS is the primary Restriction Enzyme gene of GSH synthesis.[NOCTORGetal.,2002]In most plants, GCS limits the synthesis rate of glutathion.The synthesis of transcription factor nrf2 participation γ-GCS heavy chain, and then affect the synthesis of GCS, it is mixed into the synthetic quantity of glutamyl cysteine on final.[JaiswalAKetal.,2004]
Conventional cross-breeding technology, Flight Mutagenesis technology, molecular mark, genomics technologies etc. can the degeneration-resistant border of selection-breeding, the crop of disease-resistant high yield.But it is long that these technology are respectively present breeding cycle, radiation hazradial bundle, the problems such as screening operation amount is big, relatively costly.
Summary of the invention
It is an object of the invention to provide a kind of Fructus Lycii glutamyl cysteine synthetase.
Second purpose of the present invention is to provide Fructus Lycii glutamyl cysteine synthetase encoding gene.
3rd purpose of the present invention is to provide containing the recombinant vector of Fructus Lycii glutamyl cysteine synthetase encoding gene, expression vector and recombinant bacterium.
4th purpose of the present invention is to provide Fructus Lycii glutamyl cysteine synthetase encoding gene and is preparing the application of transgenic plant.
Technical scheme is summarized as follows:
A kind of Fructus Lycii glutamyl cysteine synthetase, is the aminoacid sequence shown in SEQIDNO.1.
Fructus Lycii glutamyl cysteine synthetase encoding gene, is the deoxynucleotide sequence shown in SEQIDNO.2.
Containing the recombinant vector of Fructus Lycii glutamyl cysteine synthetase encoding gene, expression vector and recombinant bacterium.
Fructus Lycii glutamyl cysteine synthetase encoding gene is preparing the application of transgenic plant, is imported in purpose plant by Fructus Lycii glutamyl cysteine synthetase encoding gene, obtains resistance to high NaCl environment stress and tolerance cadmium ion transgenic plant.
Described purpose plant is corn and soybean, Oryza sativa L., Semen arachidis hypogaeae, Helianthi, Rhizoma Solani tuber osi, Fructus Hordei Vulgaris, Semen Tritici aestivi, Cotton Gossypii, Chinese rose, Lisianthus, Bulbus Lilii, An Zuhua, iris, Euonymus japonicus or fragrant flower Chinese scholar tree.
Advantages of the present invention: the Fructus Lycii glutamyl cysteine synthetase encoding gene of the present invention imports in purpose plant, obtains the transgenic plant that tolerance cadmium ion ability is high and salt resistance ability is high.
Accompanying drawing explanation
Fig. 1. with P1, P2 and P3, P4 primer electrophoretogram to the PCR primer (a) of Fructus Lycii cDNA He (b) respectively.
Fig. 2 .LcGCS connects the PCR primer electrophoretogram of pMD18-T carrier.
Fig. 3 .pMD18-T-LcGCS carrier schematic diagram.
Fig. 4 .LcGCS connects the PCR the result of pET-28a carrier.
Fig. 5 pET28a-LcGCS protein expression electrophoretogram in e. coli bl21.
Fig. 6 .pCAMBIA2300-LcGCS digestion verification electrophoretogram.
Fig. 7 .pCAMBIA2300-LcGCS carrier schematic diagram.
Fig. 8. transgene tobacco genome and wild type (WT) PCR primer electrophoretogram.
Fig. 9. transgenic corns, Semen sojae atricolor, Oryza sativa L., Semen arachidis hypogaeae, Helianthi, Rhizoma Solani tuber osi, Cotton Gossypii, Fructus Hordei Vulgaris, Wheat volatiles PCR primer electrophoretogram.
Figure 10. transgenic Chinese rose, Lisianthus, Bulbus Lilii, iris Genomic PCR products electrophoretogram.
Figure 11. transgenic Euonymus japonicus, fragrant flower Chinese scholar tree Genomic PCR products electrophoretogram.
Detailed description of the invention
Below by specific embodiment, the present invention is further illustrated.
Embodiment 1
The clone of Fructus Lycii glutamyl cysteine synthetase LcGCS gene
Utilize Trizol reagent, total serum IgE is extracted from fresh Fructus Lycii (China Fructus Lycii) blade of 100mg, adopt Transgentransecriptone-stepgDNAremovalandcDNAsynthesissu permix test kit, with Fructus Lycii total serum IgE for template, OligodT18For primer, synthesis cDNA the 1st chain under the effect of AMV reverse transcriptase.
nullThe Unigene sequential design according to Fructus Lycii transcript profile data base forward primer LcGCSL-F:5 ' GTGACACCCAATTGATTGCTATA3 ' shown in SEQIDNO.3 and shown in SEQIDNO.4 downstream primer LcGCSL-R:5 ' CTGTTGTGGGGAAAATGTATGTC3 '，The long segment of glutamyl cysteine synthetase LcGCS gene in pcr amplification Fructus Lycii，PCR primer electrophoretogram，See Fig. 1. (a)，Second time PCR is carried out as template after diluting 100 times using this PCR primer again，The second time forward primer P1:5 ' CTTACTGAGCCAAGGCACAAAG3 ' shown in SEQIDNO.5 of nest-type PRC and the primer P2:5 ' GCTGAATTGAAATGCACTCTCTC3 ' shown in SEQIDNO.6 amplification obtains complete gene order，PCR primer is shown in Fig. 1. (b).Utilize Tian Gen company common DNA product purification kit that PCR product is purified after reaction, operate according to test kit description, obtain the LcGCSPCR fragment of purification.
PCR primer sequence is verified, it is thus achieved that length is 1611bp sequence.GCS deoxynucleotide sequence comparison with other species, it has been found that be ORF (openreadingframe) sequence of LcGCS.Its protein sequence is predicted, as shown in SEQIDNO.1 in the ORFFinder website of NCBI.Its deoxynucleotide sequence is such as shown in SEQIDNO.2
Embodiment 2
The building process of recombinant vector pMD18-T-LcGCS
LcGCS gene shown in sequence table SEQ IDNO.2 is connected with pMD18-T carrier,
Reaction condition: 16 DEG C, 30min.Connect product Transformed E .ColiTop10.Select white colony, bacterium colony PCR method confirms the length scale of Insert Fragment in carrier T, such as Fig. 2, consistent with expection, this carrier is sent to the order-checking of Hua Da genome company, we obtain this gene deoxynucleotide sequence of 1611bp, blast is carried out at NCBI, with Fructus Lycopersici esculenti, the homology such as Rhizoma Solani tuber osi is high, it was shown that for this gene clone success.LcGCS deoxynucleotide sequence and pMD18-T sequence assembly are assembled into cloning vehicle pMD18-T-LcGCS, as shown in Figure 3.
Embodiment 3
The building process of escherichia coli RT-PCR expression vector pET28a-LcGCS
First with pMD18-T-LcGCS plasmid for template, P3, the P4 that SEQIDNO.7, SEQIDNO.8 are respectively shown in is respectively upstream and downstream primer, expands LcGCS gene.(SEQIDNO.7:CGCGGATCCATGGCCTTGATGTCTCAGGC) (SEQIDNO.8:ACGCGTCGACTCAGTAGAGAAGCTCCTCAAAGAC)
Its reaction condition is: 94 DEG C, 4min；(94 DEG C, 30Sec；56 DEG C, 30Sec；72 DEG C, lmin50Sec) 32cycles；72 DEG C, 8min.
P3 has BamHI restriction enzyme site (GGATCC), P4 has SalI restriction enzyme site (GTCGAC), then PCR primer and pET28a empty carrier plasmid are respectively with BamHI and SalI double digestion, the digestion products of the two is connected, connect product Transformed E .ColiDH5 α, coat on the LB flat board containing 200mg/Lkana resistance, 37 DEG C of cultivations.After 12h, picking list bacterium colony carries out bacterium colony PCR checking, such as Fig. 4, by bacterium positive for bacterium colony PCR checking, shakes bacterium and extracts plasmid, and enzyme action is identified and obtained purpose band, finally send the order-checking of Hua Da gene sequencing company, and it is correct that result shows that carrier pET28a-LcGCS builds.
The prokaryotic expression carrier pET28a-LcGCS built is converted e. coli bl21.Carry out protein expression checking, obtain LcGCS expressing protein such as Fig. 5 of expection size (58.6KD).Be followed successively by tropina after pET28a empty carrier bacterium does not induce tropina, the induction of pET28a empty carrier bacterium from left to right, pET28a-LcGCS expression vector bacterium does not induce tropina, pET28a-LcGCS expression vector bacterium induction tropina
Embodiment 4
Microorganism, plant eukaryotic recombinant expression carrier pCAMBIA2300-LcGCS structure
First with pMD18-T-LcGCS plasmid for template, P3, P4 is upstream and downstream primer for (deletion) respectively, expands LcGCS gene, and its reaction condition is: 94 DEG C, 4min；(94 DEG C, 30Sec；56 DEG C, 30Sec；72 DEG C, lmin50Sec) 32cycles；72 DEG C, 8min.P3 has BamHI restriction enzyme site (GGATCC), P4 has SalI restriction enzyme site (GTCGAC), then PCR primer and pCAMBIA2300 empty carrier plasmid are respectively with BamHI and SalI double digestion, the digestion products of the two is connected, connect product Transformed E .ColiDH5 α, coat on the LB flat board containing 100mg/L concentration kana resistance.37 DEG C of cultivations, after 12h, picking list bacterium colony carries out bacterium colony PCR checking, by bacterium positive for bacterium colony PCR checking, shake bacterium and extract plasmid, enzyme action is identified and is obtained purpose band, such as Fig. 6 (being followed successively by pCAMBIA2300-LcGCS carrier, pCAMBIA2300-LcGCS digestion products from left to right), finally sending the order-checking of Hua Da gene sequencing company, it is correct that result shows that carrier pCAMBIA2300-LcGCS builds.LcGCS deoxynucleotide sequence and pCAMBIA2300 sequence assembly are assembled into expression vector pCAMBIA2300-LcGCS, as shown in Figure 7.
Embodiment 5
Structure for the Agrobacterium engineering strain C58:pCAMBIA2300-LcGCS of plant transgene.
Prepared by Agrobacterium competent cell
1. by mono-for Agrobacterium C58 colony inoculation in 5mLYEP fluid medium, 28 DEG C, 180r/min shaken cultivation 36h.
2. proceeding in l00mLYEP fluid medium by above-mentioned bacterium solution 2ml, 28 DEG C, 180r/min, shaken cultivation is to (OD600Value is about 0.4-0.5).
3., after ice bath 30min, 4 DEG C, 5000r/min is centrifuged l0min, collects thalline, is resuspended in the ddH of 20mL pre-cooling2In O.
4.4 DEG C, 5000r/min is centrifuged 10min, collects thalline, is resuspended in 15% glycerol of pre-cooling, often after pipe 200 μ L liquid nitrogen flash freezer, be stored in immediately-80 DEG C standby.
Plant expression vector electroporated
1. the C58 competent cell that-80 DEG C are taken out is placed on ice so that it is slowly melt；
2. add 2-5 μ LpCAMBIA2300-LcGCS plasmid, mixing, ice bath 5min；
3. it is transferred in the electric shock cup of pre-cooling；
4. electroporated instrument parameter is set: 1500V, 5-6ms, electroporated；
5. room temperature adds 500 μ LYEP fluid mediums after standing 2min, 28 DEG C, 180r/min shaken cultivation 3-4h；
6. the centrifugal 10min of room temperature 4000r/min, sucking-off 400 μ L of supernatant liquid, mix remaining bacterium solution, coat on the YEB flat board containing 100mg/L kanamycin and 100mg/L rifampicin resistance, is inverted flat board, 28 DEG C of cultivations, 36-48h, until seeing single bacterium colony clearly.
7. picking list bacterium colony, bacterium colony PCR verifies.
Embodiment 6
Agriculture bacillus mediated Nicotiana tabacum L. genetic transformation
The aseptic culture of tobacco seedling: select full, healthy tobacco seed, with containing 25%NaOCl (effective chlorine >=10%) 20ml, the aqueous solution of concentrated hydrochloric acid 4ml, suffocating sterilization 1.5h, seed is placed in MS culture medium, 16h/8h light dark cycles, 25 ± 1 DEG C of cultivations.
The culture medium that this experiment is used is as shown in the table
The Agrobacterium-mediated Transformation of Nicotiana tabacum L.
Infect the preparation of bacterium solution
1. picking positive Agrobacterium list bacterium colony, is inoculated in the 5ml YEP fluid medium containing 100mg/L kanamycin and 100mg/L rifampicin, in 28 DEG C, 24h cultivated by the shaking table of 200r/min.
2. next day, take 3ml bacterium solution, be inoculated in the 50mlYEP fluid medium containing 100mg/L kanamycin and 100mg/L rifampicin, when bacterium solution is in vigorous period (OD600=0.4-0.6) time, bacterium solution is poured into 50ml centrifuge tube, at 5000r/min, at 4 DEG C, centrifugal 10min, abandons supernatant, collects thalline.Resuspended with the MS fluid medium of equivalent, make OD600=0.9～1.
Outer implant is contaminated
1. tobacco leaf is removed master pulse and limb edge, then blade is cut into 0.5cm × 0.5cm size, immerse the Agrobacterium bacterium solution prepared, soaking 15-20min, shake 2-3 time, makes blade be fully contacted bacterium solution therebetween, take out blade, by the sterile water wash containing kanamycin and with the aseptic filter paper unnecessary liquid of exhaustion, blade face down, blade back be inoculated in common training culture medium upward, about 25 DEG C light culture 3 days.
2. being transferred in screening culture medium by blade, within about 20 days, change a subculture, induction of resistance bud produces, and when resistant buds grows to about 1cm from callus, cuts resistant buds wound healing, is inoculated in resistance seedling rooting culture medium.
Transgenic seedling is transplanted
Root growth is good, vitality is vigorous Nicotiana tabacum L. tissue cultured seedling takes out from tissue culture bottle, with tap water culture medium (reducing root system damage) as far as possible, it is planted in perlite, in the compost of Vermiculitum and fertile soil (1:2:2), cover film moisturizing, ventilative, at 25 DEG C, cultivate 7-10 days, then open thin film, regularly water, apply fertilizer, and be transferred in greenhouse.
Embodiment 7
Transgene tobacco Molecular Detection and salt-resistance detection
The PCR detection of transgene tobacco genomic DNA: 1.CTAB method extracts T1 for Nicotiana tabacum L. STb gene；2. detecting with genomic DNA for template performing PCR, primer is P3, P4.
Take above-mentioned PCR primer 5 μ l and carry out electrophoresis detection, see Fig. 8, illustrate that LcGCS gene successfully proceeds to Nicotiana tabacum L..
At greenhouse normal growth, 5-7cm transgene tobacco Seedling and wild type control Seedling, water the water of NaCl containing 300mmol/L and 500mmol/L concentration respectively, greenhouse keeps the humidity of 85%.Detection finds, under the growth conditions of 300mmol/LNaCI, WT lines growth phase is to slowly, Biomass is relatively fewer, and blade is partially yellow, and transgene tobacco can normal growth, Biomass is big, and blade is greener, it is possible to blossom and bear fruit, when the NaCl of 500mmol/L concentration, after growing one week, wild-type tobacco yellow leaf, wilt, tissue necrosis, it is impossible to normal growth.Although the growth of transgene tobacco is also affected by a degree of suppression, but obvious unlike wild-type tobacco, substantially can normal growth.
At greenhouse normal growth, choose the consistent transgene tobacco Seedling of growth and wild type control Seedling, cultivate respectively at the CdCl containing 200 μm of ol/L and 300 μm of ol/L concentration21/10hoagland nutritional solution in, in greenhouse keep 85% humidity.After 4 weeks, detection finds: 200 μm of ol/L process group transgene tobacco cross phase true leaves turn yellow, but top is bud green, and root system compared with normal plant is short and thick, but can grow, but non-transgenic tobacco blade and stem all turn yellow, and root system is very short, and plant is downgraded serious, withered；300 μm of ol/L process groups, transgenic group tobacco growing is suppressed, yellow leaf, and root system is short and thick, and color browning is yellow.
Embodiment 8
This laboratory passes through seed embryo growing point gene brush Wicresoft infestation method (Rong Fei, Wang Gang, Ji Jing, Deng. utilize " Wicresoft's brush " method to obtain resistance glyphosate genetically engineered soybean [J]. Soybean Science, 2015, 34 (2) .) to Semen Maydis, Semen sojae atricolor, Oryza sativa L., Semen arachidis hypogaeae, Helianthi, Rhizoma Solani tuber osi, Fructus Hordei Vulgaris, Semen Tritici aestivi, crop maturity embryo transgenic (LcGCS) such as Cotton Gossypii, it is successfully obtained transgenic corns, Semen sojae atricolor, Oryza sativa L., Semen arachidis hypogaeae, Helianthi, Rhizoma Solani tuber osi, Fructus Hordei Vulgaris, Semen Tritici aestivi, cotton plants, Genomic PCR electrophoretogram (such as Fig. 9), it is followed successively by Semen Maydis from left to right, Semen sojae atricolor, Oryza sativa L., Semen arachidis hypogaeae, Helianthi, Rhizoma Solani tuber osi, Fructus Hordei Vulgaris, Semen Tritici aestivi, the genome of Cotton Gossypii and the PCR primer of carrier 2300-LcGCS.
At greenhouse normal growth, 5-7 true leaf transgenic seedlings and wild type control Seedling, water the water of NaCl containing 300mmol/L and 500mmol/L concentration respectively, greenhouse keeps the humidity of 85%.Detection finds, under the growth conditions of 300mmol/LNaCl, WT lines growth phase is to slowly, Biomass is relatively fewer, and blade is partially yellow, and transgenic seedlings can normal growth, Biomass is big, and blade is greener, it is possible to blossom and bear fruit, when the NaCl of 500mmol/L concentration, after growing one week, wild type seedlings yellow leaf, wilt, tissue necrosis, it is impossible to normal growth.Although the growth of transgenic seedlings is also affected by a degree of suppression, but obvious unlike wild type seedlings, substantially can normal growth.
At greenhouse normal growth, choose the consistent transgenic seedlings of growth and wild type control Seedling, cultivate respectively at the CdCl containing 200 μm of ol/L and 300 μm of ol/L concentration21/10hoagland nutritional solution in, in greenhouse keep 85% humidity.After 4 weeks, detection finds: 200 μm of ol/L process group transgenic seedlings cross phase true leaves turn yellow, but top is bud green, and root system compared with normal plant is short and thick, but can grow, but non-transgenic seedlings blade and stem all turn yellow, and root system is very short, and plant is downgraded serious, withered；300 μm of ol/L process groups, transgenic group growth of seedling is suppressed, yellow leaf, and root system is short and thick, and color browning is yellow.
Embodiment 9
This laboratory utilizes Agrobacterium infestation method that the flowers such as Chinese rose, Lisianthus, Bulbus Lilii, iris are carried out axillalry bud or stem scale Wicresoft's method transgenic (LcGCS), obtain transgenic Chinese rose, Lisianthus, Bulbus Lilii, Phalaenopsis plants, Genomic PCR electrophoretogram such as Figure 10, is followed successively by the PCR primer of Chinese rose, Lisianthus, Bulbus Lilii, the genome of iris and carrier 2300-LcGCS from left to right.
At greenhouse normal growth, 5-7cm transgenic seedlings and wild type control Seedling, water the water of NaCl containing 300mmol/L and 500mmol/L concentration respectively, greenhouse keeps the humidity of 85%.Detection finds, under the growth conditions of 300mmol/LNaCl, WT lines growth phase is to slowly, blade is partially yellow, it is impossible to normally bloom, and transgenic seedlings can normal growth, blade is greener, can bloom, when the NaCl of 500mmol/L concentration, after growing one week, wild type seedlings yellow leaf, wilt, tissue necrosis, it is impossible to normal growth.Although the growth of transgenic seedlings is also affected by a degree of suppression, but obvious unlike wild type seedlings, substantially can bloom by normal growth.
At greenhouse normal growth, choose the consistent transgenic seedling of growth and wild type control Seedling, cultivate respectively at the CdCl containing 200 μm of ol/L and 300 μm of ol/L concentration21/10hoagland nutritional solution in, in greenhouse keep 85% humidity.After 4 weeks, detection finds: 200 μm of ol/L process group transgenic seedling cross phase true leaves turn yellow, but top is bud green, and root system compared with normal plant is short and thick, but can grow, but non-transgenic seedling leaf and stem all turn yellow, and root system is very short, and plant is downgraded serious, withered；300 μm of ol/L process groups, transgenic seedling growth is suppressed, yellow leaf, and root system is short and thick, and color browning is yellow.
Embodiment 10
This laboratory utilizes Agrobacterium infestation method that the arbor trees such as Euonymus japonicus, fragrant flower Chinese scholar tree is carried out transgenic (LcGCS), obtain transgenic Euonymus japonicus, fragrant flower Chinese scholar tree plant, Genomic PCR electrophoretogram (such as Figure 11), is followed successively by the PCR primer of Euonymus japonicus, the genome of fragrant flower Chinese scholar tree and carrier 2300-LcGCS from left to right.
At transgenic seedling and the wild type control Seedling of greenhouse normal growth, water the water of NaCl containing 300mmol/L and 500mmol/L concentration respectively, greenhouse keeps the humidity of 85%.Detection finds, under the growth conditions of 300mmol/LNaCl, WT lines growth phase is to slowly, and blade is partially yellow.And transgenic seedling can normal growth, blade is greener.When the NaCl of 500mmol/L concentration, after growing one week, wild type seedling leaf turns to be yellow, and wilts, tissue necrosis, it is impossible to normal growth.Although the growth turning base seedling is also affected by a degree of suppression, but obvious unlike wild type seedlings, substantially can normal growth.
At greenhouse normal growth, choose the consistent transgenic seedling of growth and wild type control Seedling, cultivate respectively at the CdCl containing 200 μm of ol/L and 300 μm of ol/L concentration21/10hoagland nutritional solution in, in greenhouse keep 85% humidity.After 4 weeks, detection finds: 200 μm of ol/L process group transgenic seedling cross phase true leaves turn yellow, but top is bud green, and root system compared with normal plant is short and thick, but can grow, but non-transgenic seedling blade and stem all turn yellow, and root system is very short, and plant is downgraded serious, withered；300 μm of ol/L process groups, the growth of transgenic seedling is suppressed, yellow leaf, and root system is short and thick, and color browning is yellow.
Claims (5)
1. a Fructus Lycii glutamyl cysteine synthetase, is characterized in that the aminoacid sequence shown in SEQIDNO.1.
2. Fructus Lycii glutamyl cysteine synthetase encoding gene, is characterized in that the deoxynucleotide sequence shown in SEQIDNO.2.
3. contain the expression vector of gene described in claim 2, recombinant vector and recombinant bacterium.
4. Fructus Lycii glutamyl cysteine synthetase encoding gene is preparing the application of transgenic plant, it is characterized in that in channel genes purpose plant described in claim 2, to obtain resistance to high NaCl environment stress and tolerance cadmium ion transgenic plant.
5. application according to claim 4, it is characterized in that described purpose plant is corn and soybean, Oryza sativa L., Semen arachidis hypogaeae, Helianthi, Rhizoma Solani tuber osi, Fructus Hordei Vulgaris, Semen Tritici aestivi, Cotton Gossypii, Chinese rose, Lisianthus, Bulbus Lilii, An Zuhua, iris, Euonymus japonicus,, fragrant flower Chinese scholar tree.
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