CN102234655A - Application of pprM gene of Deinococcus radiodurans R1 in improvement of drought-resistant traits of plants - Google Patents
Application of pprM gene of Deinococcus radiodurans R1 in improvement of drought-resistant traits of plants Download PDFInfo
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Abstract
The invention discovers that pprM gene in Deinococcus radiodurans R1 can enhance the drought resistance of prokaryotes and plants. Recombinant vectors comprising the gene are constructed and respectively introduced into procaryotic and enkaryotic host cells. Experiments prove that after the pprM gene is expressed in prokaryotic host cells and plants, the resistance to drought environment can be enhanced.
Description
Technical field
The present invention relates to the new function of the different coccus R1 of radiation hardness pprM gene (Deinococcus radiodurans R1 pprM), be specifically related to this pprM gene in the purposes of improvement plant to aspects such as drought stress resistances.
Background technology
The different coccus Deinococcus of radiation hardness radiodurans R1 is a kind of rhodobacterium that people such as Anderson in 1956 find from the can of handling through radiation sterilization.
PprM gene (DR_0907 among the Deinococcus radiodurans R1, GeneID:1797366) note is a cold shock protein, find that people such as Hirofumi Ohba in 2009 this gene can depend on the relevant albumen (Ohba of a series of dna damage reparations such as PprI regulation and control PprA, H.Satoh, et al., 2009) participate in dna damage reparation after the ionizing rays, this gene also is the key gene that Deinococcus radiodurans R1 has radiation resistance.
Have synergetic property although it is generally acknowledged radiation resistance and drought stress resistance on evolving, at present relevant Deinococcusradiodurans R1 pprM gene is for the research that improves the plant drought performance, and not seeing has report.
Summary of the invention
The objective of the invention is from D.radiodurans R1 genome, to find to strengthen the gene of plant arid resistance.And, make it obtain the ability of opposing arid with this gene transferred plant.
The inventor finds, (DR_0907 GeneID:1797366) has the function of drought resisting to Deinococcus radiodurans R1 pprM gene, can be used for cultivating the plant of drought-resistant character.
The present invention has carried out following work:
1. amplify D.radiodurans R1pprM gene (DR_0907) by PCR from D.radiodurans R1 (DSM 20539) strain gene group, gene order number is: GeneID:1797366.Its size is 402bp, and it is cloned on carrier pGEMT-easy, has made up the recombinant plasmid pGEMT-pprM that contains complete pprM gene;
2. the groEL promotor that can all work in intestinal bacteria and the different coccus of radiation hardness is connected on the above-mentioned pGEMT-pprM recombinant plasmid, makes up to have the recombinant plasmid pGEMT-pprMG that groEL contains complete pprM gene;
3. the recombinant plasmid pGEMT-pprMG that will import the pprM gene changes in the acceptor e. coli jm109, obtains engineering strain JM_pprM;
Through experiment confirm, this bacterial strain has resists 4M NaCl and the ballistic ability of high density (3M) sorbyl alcohol (Sorbitol) (because of Sorbitol has water absorbability, the environment that the Sorbitol solution of high density can simulating drought sees embodiment 2 for details) respectively;
4. D.radiodurans R1 pprM gene (DR_0907) is connected among the plant expression vector pBI121, makes up recombinant plasmid pBI121-pprM with pprM gene;
5. the pBI121-pprM recombinant plasmid is imported in the rape of the arid of tool not resistance, acquisition can tolerate the rape plant of arid;
Experimental result shows that the transgene rape of the D.radiodurans R1 pprM gene that contains gained of the present invention has obtained the stronger tolerance to arid (seeing embodiment 5,6 for details, 7).
Description of drawings:
Fig. 1 is the PCR product that contains D.radiodurans R1 pprM gene (DR_0907) sequence;
Fig. 2 is the construction of prokaryotic expression vector checking electrophoretogram that contains D.radiodurans R1 pprM gene (DR_0907) and groEL promotor;
Fig. 3 is the checking electrophoretogram that contains the plant expression vector construction of D.radiodurans R1 pprM gene (DR_0907) sequence;
Fig. 4 is containing empty carrier and containing the colibacillary upgrowth situation of the prokaryotic expression carrier of D.radiodurans R1 pprM gene (DR_0907) sequence before shock test, wherein:
A is intestinal bacteria JM 109 bacterial strains that contain empty expression vector;
B is the e. coli jm109 bacterial strain that contains D.radiodurans R1 pprM gene (DR_0907) sequence expression vector;
Fig. 5 contains the prokaryotic expression carrier of D.radiodurans R1 pprM gene (DR_0907) sequence and intestinal bacteria (E.coli) upgrowth situation in the substratum that contains the 3M sorbyl alcohol of empty carrier, proves that D.radiodurans R1 pprM gene (DR_0907) can strengthen the function that intestinal bacteria have drought resisting.Bacterial strain among the figure is as follows:
A is the e. coli jm109 bacterial strain that contains empty expression vector;
B is the e. coli jm109 bacterial strain that contains D.radiodurans R1 pprM gene (DR_0907) sequence expression vector.
Fig. 6 contains intestinal bacteria upgrowth situation in the substratum that contains 4M NaCl of D.radiodurans R1 pprM gene (DR_0907) sequence expression vector and empty carrier, proves that D.radiodurans R1 pprM gene (DR_0907) has the salt tolerant resistance.Bacterial strain among the figure is as follows:
A is the e. coli jm109 bacterial strain that contains empty expression vector;
B is the e. coli jm109 bacterial strain that contains D.radiodurans R1 pprM gene (DR_0907) sequence expression vector.
Fig. 7 is the RT-PCR checking of transgene rape.
Fig. 8 is the state of rape before processing that changes the pprM gene.
Fig. 9 is the state of negative control (not genetically modified rape) before processing.
Figure 10 is the rape that changes the pprM gene stop the to water state in 1 week.
Figure 11 is negative control (not genetically modified rape) stop the to water state in 1 week.
Figure 12 is the rape that changes the pprM gene stop the to water state in 2 weeks.
Figure 13 is negative control (not genetically modified rape) stop the to water state in 2 weeks.
Figure 14 is transgene rape and the membrane permeability contrast of wild-type rape plant tissue cell behind the drought stress, and wherein ordinate zou shows ion permeability.
Figure 15 is transgene rape and the contrast of wild-type rape leaf mda content behind the drought stress, and wherein ordinate zou shows mda content.
Embodiment
The plasmid of being lifted in following examples, bacterial strain just are used for the present invention is described in further detail, and flesh and blood of the present invention are not limited.All unreceipted concrete experiment conditions, be according to normal condition well known to those skilled in the art, for example the Sambrook equimolecular is cloned: laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.
Plasmid, the bacterium source lifted among the embodiment are as follows:
Cloning vector pGEMT-easy: be Promrga company commercially available prod;
Plant expression vector pBI121: be Clontech company commercially available prod;
Intestinal bacteria JM 109: be Beijing Quan Shi King Company commercially available prod.Agrobacterium EHA 105 is preserved by this laboratory
Embodiment 1 expression of D.radiodurans R1 pprM gene order in intestinal bacteria
According to the design of the pprM gene order in the D.radiodurans R1 genome of having announced one couple of PCR Auele Specific Primer, the complete nucleotide sequence of amplification from the D.radioduransR1 genomic dna.
Up?5′AGCGACTAGT?GCCAAAT?ATGTGCCGAGTTTC?3′；Down?5′ATCCCATATGTTACCAGCGGTCGTCGCGGC3′。From the genome of D.radiodurans R1, amplify sequence SEQ ID NO:1 by PCR method, condition: 95 ℃ of 10min, [95 ℃ of 30sec, 55 ℃ of 30sec, 72 ℃ of 45sec] 30 circulations, 72 ℃ of 10min, the PCR product is cloned on carrier pGEMT-easy after glue reclaims, called after pGEMT-pprM, the groEL promotor that will come from D.radiodurans R1 is then also linked on this carrier, makes up coli expression carrier pGEMT-pprMG, with this expression vector transformed into escherichia coli JM109, through PCR, enzyme is cut, correct (the seeing Fig. 1,2) of three aspects that check order checking insertion sequence is with this bacterial strain called after JM_pprM.
The E.coli JM109 called after JM_pGEMT-G that will contain the pGEMT-easy+groEL empty plasmid.
Embodiment 2 contains the engineering strain raising arid of D.radiodurans R1 pprM gene and the tolerance experiment of salt
One, experimental technique
1, the intestinal bacteria with two reorganization of gained among the embodiment 1 are inoculated in the 20mL LB liquid nutrient medium, behind the shake-flask culture 3-4h, transfer again in the LB of 100mL liquid nutrient medium, keep the unanimity of inoculum size as far as possible, be cultured between the OD0.3-0.4, transfer to OD value unanimity as far as possible.
2, get the bacterium liquid of 10mL centrifugal after, in the sorbitol solution of isopyknic 4M NaCl salts solution and 3M, impact 2h, each sample is used aseptic deionized water proportional diluted to 10 immediately
-4, get 10 μ L points at LB solid culture primary surface, through 37 ℃ of cultivation 16h, observe also and take a picture.The Sorbitol Powder of 4M NaCl salts solution and 3M is hypertonic solution, can coerce by simulating drought.
Two, experimental result
As can be seen from Figure 4, the JM_pprM bacterial strain and the JM_pGEMT-G strain growth state basically identical that contains empty plasmid that contain D.radiodurans R1 pprM gene (DR_0907) before the impact; After 4M NaCl salts solution and 3M sorbyl alcohol impact, the JM_pprM strain growth that contains D.radiodurans R1 pprM gene (DR_0907) in order, the bacterium colony number is obviously more than the JM_pGEMT-G bacterial strain that only contains empty plasmid (seeing Fig. 4,5,6).
Three, conclusion
D.radiodurans R1 pprM gene can help coercing of prokaryotic organism tolerance arid and salt.
Embodiment 3 D.radiodurans R1 pprM genes carry out the evaluation of eukaryotic cell expression and transfer-gen plant in rape cell
(1) contains the structure of goal gene plant expression vector
Design the primer that the pprM gene is used for expression of plants according to D.radiodurans R1 genome sequence, upstream primer adds Bam HI, and downstream primer adds two restriction enzyme sites of Sac I, and initiator codon is changed into atg from gtg.Primer sequence is as follows: DR0907 PLANT UP:CGGCGGATCC ATGTGCCGAGTTTCGATTGT;
DR0907?PLANT?DOWN：ATTAGAGCTC?TTACCAGCGGTCGTCGCGGC。
And the upstream has Bam HI restriction enzyme site, the downstream has Sac I restriction enzyme site, pprM (DR_0907) gene is increased from genome, fragment is through Bam HI, Sac I double digestion glue is connected on the plant expression vector pBI 121 of same double digestion after reclaiming, and constructs and contains complete DR0907 expression carrier pBI 121-pprM (see figure 3).
(2) agrobacterium mediation converted rape experiment
1. the competent preparation of agrobacterium tumefaciens EHA105:
1) picking list bacterium colony is inoculated in 5mL YEB liquid nutrient medium (containing Rifampin Rif 50mg/L), and 28 ℃, the 250rpm shaking culture is spent the night;
2) get 2mL bacterium liquid, add in the 50mL YEB liquid nutrient medium (containing Rif 50mg/L), 28 ℃, the 250rpm shaking culture is to OD
600About about 0.6;
3) bacterium liquid is gone in the aseptic centrifuge tube of 50mL ice bath 30min, the centrifugal 5min of 5000 * g;
4) abandon supernatant, precipitation 2mL 20mM CaCl
2Heavily floating, every part 100 μ L branch installs in the 1.5mL centrifuge tube, preserves standby in the liquid nitrogen.
2. recombinant plasmid dna changes Agrobacterium over to:
1) the pBI-pprM plasmid DNA with about 1 μ g joins in the 100 μ L EHA105 competent cells mixing, ice bath 5min respectively;
2) centrifuge tube is put freezing 8min in the liquid nitrogen, gone to temperature bath 5min in 37 ℃ of water-baths rapidly;
3) add 1mL YEB liquid nutrient medium, 250rpm recovery 4～5h on 28 ℃ of shaking tables;
4) get an amount of bacterium liquid and be applied on the YEB solid medium that contains Rifampin (Rif) 50mg/L and kantlex (Kan) 100mg/L, put 28 ℃ and cultivate 24～48h.
3. the extraction of the plasmid of Agrobacterium
1) picking colony (YEB: Tryptones 5g/L, yeast extract 1g/L, sucrose 5g/L, sal epsom 0.5g/L) in the YEB liquid nutrient medium that contains Rifampin (Rif) 50mg/L and kantlex (Kan) 100mg/L, 28 ℃, 250rpm shaking culture 20h;
2) get 1.5mL bacterium liquid centrifugal after, abandon supernatant, with STE solution (Tris-HCI 10mM pH8.0, NaCl 10mM, EDTA 10mM pH 8.0) washed twice, abandon supernatant, add solution I (50mM glucose, the 25mM Tris-HClpH8.0 of precooling, 10mM EDTApH 8.0, RNaseA 100 μ g/mL) 300 μ L aspirate mixing repeatedly with pipettor, and room temperature leaves standstill 10min;
3) (solution II of the fresh configuration of adding turns upside down mixing several times for 0.2M NaOH, 1%SDS) 600 μ L;
4) solution III (5M potassium acetate pH 5.2, Glacial acetic acid 11.5mL adds water to 100mL) the 450 μ L of adding precooling, abundant mixing, ice bath 5min, 4 ℃ of centrifugal 5min of 12000 * g get the isopropanol precipitating of supernatant with 0.6 times of volume;
5) after precipitation is dissolved with TE (pH 8.0 for Tris-HCl 10mmol/L, 1mmol/LEDTA), through PCR, BamHI, the checking of Sac I double digestion.
4. the preparation of rape aseptic seedling and agriculture bacillus mediated pprM gene genetic transform
1) cultivation of rape aseptic seedling
Swede type rape (Brassica napus L.) (84100-18) seed soaks 10min with aqua sterilisa on Bechtop, 10% NaClO sterilization 12min, again with 0.1% mercuric chloride solution sterilization 7-8min, with sterilization washing 3-5 time, and the Semen Brassicae campestris of sterilization is placed in equably on the pre-culture medium of no hormone (7.5% agar).Illumination cultivation, 4-5d rape in age seedling is suitable for genetic transformation most.
2) the pre-cultivation
The scissors of alcohol with 75% and high-temperature sterilization sterilization is cut cotyledon and vegetative point, and clip rape aseptic seedling next-door neighbour vegetative point is the long hypocotyl of about 0.5cm down, places pre-culture medium (MS+6-BA 2mg/L+2,4-D 1 mg/L+AgNO
32.5mg/L+AS 19.62mg/L), 2d is cultivated in illumination in advance.
3) cultivation of Agrobacterium
In 50mL LB liquid nutrient medium (containing kantlex 100mg/L, Rifampin 50mg/L), add 0.1mLpprM-EHA 105 bacterium liquid, about 28 ℃ of following 220rpm shaking culture 16h, the centrifugal 10min of 4000 * g under the room temperature, abandon supernatant liquor, thalline suspends with the MS liquid nutrient medium (containing AS 100 μ mol/L) of sterilization, the 5-20 that is diluted to original volume doubly at 28 ℃ of following 220rpm shaking culture 1h, makes the OD of bacterium liquid
600Reach about 0.5.
4) cultivate altogether
The pre-hypocotyl of cultivating 2d is put into bacterium liquid contaminate 1min, the medication spoon is pulled hypocotyl out, be placed on the thieving paper of sterilization, blot the unnecessary bacterium liquid on the hypocotyl, be put into again on the pre-culture medium that is coated with sterilization filter paper, seal culture dish with sealing film, about 25 ℃, cultivate about 2d (the dark cultivation) altogether in the dark place.
5) inducing culture
The hypocotyl of cultivating 2d altogether is put into inducing culture (MS+6-BA 2mg/L+AgNO
32.5mg/L) on, seal culture dish with sealing film, 25 ℃ of left and right sides illumination cultivation, per two all subcultures once, until growing the callus that expands.
6) select to cultivate
The callus that expands is transferred to screening culture medium (MS+6-BA 2mg/L+AgNO
32.5mg/L+Kan 100mg/L), seal culture dish with sealing film, 25 ℃ of left and right sides illumination cultivation, per two all subcultures once, until growing seedling.
7) root culture
When high, they are separated from callus to 1-2cm when young shoot is long, transfer to that (1/2MS+NAA 0.5mg/L+Kan 25mg/L) carries out root culture on the root media.Under the antibiotic-screening condition, about 87% bud is at two weeks back formation root.
8) hardening and transplanting
Seedling after taking root is long to 5-6cm when high, and half opens wide the culturing bottle lid, carries out hardening; Treat that seedling adapts to after the external environment, transfer in the vermiculite of indoor sterilization plantation and cultivate, and water with the 1/2MS nutrient solution.When growth of seedling during, sprigging is grown in earth, and then carry out next step experiment to 7-9cm.
Embodiment 4 utilizes the RT-PCR method to detect the expression of pprM gene in the transgene rape plant
The extraction of the total RNA of 1 plant
The total RNA extraction agent of the centrifugal a small amount of plant of the post box that uses Shanghai China Shun Bioisystech Co., Ltd to produce extracts the RNA of the commentaries on classics pprM vector for transgenic rape that obtains, and handles DNA with DNase I.
2RT-PCR
Test kit (the DyNAmo of NEB company is used in reverse transcription
TMCDNA Synthesis Kit).The cDNA that obtains with reverse transcription is a template, utilizes primer DR0907 PLANT UP:CGGCGGATCCATGTGCCGAGTTTCGATTGT;
DR0907 PLANT DOWN:ATTAGAGCTC TTACCAGCGGTCGTCGCGGC carries out the PCR reaction, and condition is: 95 ℃ of 10min, [95 ℃ of 30sec, 55 ℃ of 30sec, 72 ℃ of 45sec] 30 circulations, 72 ℃ of 10min.
The PCR product after agarose gel electrophoresis detects, sequence verification.The (see figure 7) in the rape that is transferred to that shows the success of pprM gene.
Embodiment 5 changes drought resisting experiment (1)---the rape plant leaf moisture determination of pprM gene plant
In view of this nucleotide sequence has been proved to be in intestinal bacteria drought stress is had resistance, further transfer-gen plant is carried out drought resistance and identify.
Do not rewater after watering sufficient nutritive medium to rape is disposable, every other day measure leaf water content.The measuring method of water content: after about 0.1g blade takes off from plant, take by weighing fresh weight (FW) immediately, then blade was immersed in the deionized water 4 hours, taking-up is inhaled with filter paper and is removed surface-moisture, take by weighing saturated fresh weight (TW), then blade is put into baking oven and dried to constant weight in 70 ℃, take by weighing dry weight (DW).
Relative water content (Relativewatercontent, RWC) (%)=(FW-DW)/(TW-DW) * 100%.
Plant is in arid treating processes, and the blade relative water content descends gradually, and along with the prolongation between the arid place, the difference of transfer-gen plant relative water content and wild-type is obvious gradually.When the wild-type plant was coerced the 4th day in dried morning, blade had begun to wilt, and water content sharply descends; Genetically modified rape drought stress is in the time of the 7th day, and blade just begins to wilt, and the blade dehydrating speed is obviously slow than wild-type; After coercing 14 days dried morning, the blade relative water content of adjoining tree drops to 49%, and the relative water content of transgenic line is between 64-68%, far above adjoining tree (seeing Fig. 8,9,10,11,12,13).
Embodiment 6 changes the drought resisting experiment (2) of pprM gene plant---transgene rape and the membrane permeability contrast of wild-type rape plant tissue cell behind the drought stress
1) experiment purpose
When plant was subjected to adverse circumstance and influences, behind high temperature or low temperature, arid, salt marsh, pathogen infection, cytolemma was destroyed, and membrane permeability increases, thereby intracellular ionogen is exosmosed, so that the specific conductivity of vegetable cell vat liquor increases.The degree that membrane permeability increases is relevant with environment stress intensity, also relevant with the power of stress resistance of plant.Like this, more same crop different varieties is coerced the increase degree of membrane permeability under the temperature in difference, can compare a little less than the strong resistance between crop.Therefore, the conductivity meter method has become an accurate and practical method identifying the crop anti-adversity power in present crop anti-adversity cultivation, the breeding.
Drought stress usually causes the vegetable cell damage, causes the cytolemma ion permeability to increase and the adipose membrane peroxidation.
2) experiment material
After changeing that the rape of the rape (preparation method is seen embodiment 2) of pprM gene and wild-type is simultaneously disposable and water sufficient nutritive medium, a do not rewater the 8th day blade of arid processing;
B is without the transgenosis of arid processing and the rape leaf of wild-type;
3) experimental technique and step
Through the rape leaf of arid processing and the rape leave agreement that contracts a film or TV play to an actor or actress 0.1g that does not early handle through overdrying, dash 3 times with distilled water, blot blade surface moisture gently with thieving paper, blade shredded place the 25mL distilled water, in vacuum drier in 0.05MPa lower pumping 20min, 25 ℃ of 2h that slowly vibrate down measure specific conductivity S1 then, measure the blank specific conductivity C1 of water simultaneously.Again sample is handled 20min in boiling water bath, be cooled to after the room temperature constant volume and measure specific conductivity S2 to 25mL, treating water is measured its blank specific conductivity C2 in the same manner.Ion permeability=(S1-C1)/(S2-C2) * 100%.
4) experimental result
In the untreated plant, ion permeability is between 12-15%; Arid was handled 8 days, and the blade cell film ion permeability of the plant of wild-type reaches 32%; The ion permeability of transfer-gen plant is 22-24%, significantly less than wild-type plant (seeing Figure 14).
Embodiment 7 changes the drought resisting experiment (3) of pprM gene plant---transgene rape and wild-type rape leaf mda content comparison behind the drought stress
1) experiment purpose
Thiobarbituricacid (TBA) colorimetry is adopted in this experiment.Its principle is: plant is under adverse environmental factor, peroxidation of membrane lipids often takes place, and (malondialdehyde MDA) is one of its product to mda, usually utilize it as the lipid peroxidation index, expression cell membrane lipid peroxidation degree and plant are to the power of adverse environmental factor reaction.
Mda reacts with thiobarbituricacid (TBA) under high temperature, acidic conditions, being formed on 532nm wavelength place has coloured trimethylammonium mixture of maximum light absorption, the specific absorbance of this mixture is 155[mmol/ (L*cm)], and have minimum light absorb at 600nm wavelength place.Can be by following formula:
A
532-A
600＝155000×C×L (1)
Calculate MDA concentration C (μ mol/L), further calculate MDA content C (μ mol/g) in the bright tissue of unit weight.A in the formula
532And A
600The absorbance of representing 532nm and 600nm wavelength place respectively.L is cuvette thickness (cm).
It is pointed out that in the plant tissue that reaction has interference effect to glucide to MDA-TBA.For eliminating this interference, through test, available following formula is eliminated the error that is caused by sucrose.
C/μmol/L＝6.45(A
532-A
600)-0.56A
450 (2)
In the formula: A
450, A
532, A
600Represent the absorbance under 450nm, 532nm and the 600nm wavelength respectively.Can directly try to achieve the concentration of MDA in the plant sample extracting solution with formula (2), further calculate its content in plant tissue.
2) experiment material
After changeing that the rape of the rape (preparation method is seen embodiment 2) of pprM gene and wild-type is simultaneously disposable and water sufficient nutritive medium, a do not rewater the 8th day blade of arid processing;
B is without the transgenosis of arid processing and the rape leaf of wild-type.
3) experimental technique and step
A gets the 0.5g rape leaf, and totally 4 parts, add 5%TCA 5mL respectively, grind back gained homogenate centrifugal 10min under 3000 * g;
B gets supernatant liquor 2mL, adds 0.67%TBA 2mL, mixes the back and boil 30min in 100 ℃ of water-baths, and is once centrifugal again after the cooling;
C measures the absorbance of supernatant liquor under 450nm, 532nm and 600nm wavelength respectively, and by formula (2) calculate MDA concentration, again the MDA content in the unit's of calculating fresh weight tissue (μ mol/g);
The otherness of two different rape MDA content of d gained The data variance analysis check.
4) experimental result
Behind the drought stress, the MDA content of transfer-gen plant also is lower than wild-type (seeing Figure 15).
Experimental results show that more than the resistance of changeing the rape of pprM gene among the Deinococcus radiodurans R1 is higher than the rape of control group far away.
Claims (6)
1.Deinococcus radiodurans R1 pprM gene (DR_0907, GeneID:1797366) purposes of cultivation drought-resistant plant.
2. the recombinant plasmid that contains the described gene of claim 1.
3. the described recombinant plasmid of claim 2 is can be at the plasmid of escherichia coli expression.
4. the described recombinant plasmid of claim 2 is the plasmid that can express in plant.
5. with the host cell of the described recombinant plasmid transformed of claim 2, comprise prokaryotic cell prokaryocyte and eukaryotic cell.
6.Deinococcus radiodurans R1 pprM gene (DR_0907, GeneID:1797366) purposes of cultivation salt-tolerant plant.
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| CN102676585A (en) * | 2012-05-14 | 2012-09-19 | 中国农业科学院生物技术研究所 | Use and utilizing method of genes dtrA and dtr B of two-component system |
| CN102807990A (en) * | 2012-08-15 | 2012-12-05 | 中国农业科学院生物技术研究所 | Application of trkA genes in deinococcus radiodurans R1 to culture of drought-resistant plant |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102559728A (en) * | 2012-02-21 | 2012-07-11 | 中国农业科学院生物技术研究所 | Application of dap gene in Deinococcus radiodurans R1 in breeding of salt tolerant plants |
| CN102676585A (en) * | 2012-05-14 | 2012-09-19 | 中国农业科学院生物技术研究所 | Use and utilizing method of genes dtrA and dtr B of two-component system |
| CN102676585B (en) * | 2012-05-14 | 2014-04-16 | 中国农业科学院生物技术研究所 | Use and utilizing method of genes dtrA and dtr B of two-component system |
| CN102807990A (en) * | 2012-08-15 | 2012-12-05 | 中国农业科学院生物技术研究所 | Application of trkA genes in deinococcus radiodurans R1 to culture of drought-resistant plant |
| CN102807990B (en) * | 2012-08-15 | 2014-05-14 | 中国农业科学院生物技术研究所 | Application of trkA genes in deinococcus radiodurans R1 to culture of drought-resistant plant |
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