CN102234655B - 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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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
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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 radiation hardness different coccus R1 pprM gene (Deinococcus radiodurans R1 pprM), be specifically related to this pprM gene improvement plant to the purposes in drought stress resistance etc.
Background technology
Radiation hardness different coccus Deinococcus radiodurans R1 is a kind of rhodobacterium that the people such as Anderson in 1956 find from the can through radiation sterilization process.
PprM gene (DR_0907 in Deinococcus radiodurans R1, GeneID:1797366) annotation is cold shock protein, find that this gene can depend on PprI and regulate and control the relevant albumen (Ohba of a series of DNA damage reparations such as PprA people such as Hirofumi Ohba in 2009, H.Satoh, et al., 2009) participate in the DNA damage reparation after ionizing rays, this gene is also the key gene that Deinococcus radiodurans R1 has radiation resistance.
Although it is generally acknowledged that radiation resistance and drought stress resistance have synergetic property on evolving, at present about Deinococcusradiodurans R1 pprM gene is for the research improving plant drought performance, and there are no report.
Summary of the invention
The object of the invention is the gene finding to strengthen plant drought resistance from D.radiodurans R1 genome.And by this gene transferred plant, make it obtain to siccocolous ability.
The present inventor finds, Deinococcus radiodurans R1 pprM gene (DR_0907, GeneID:1797366) has the function of drought resisting, can be used for the plant cultivating drought-resistant character.
Invention has been 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, is cloned on carrier pGEMT-easy, constructs the recombinant plasmid pGEMT-pprM containing complete pprM gene;
2. the groEL promotor that all can work in intestinal bacteria and the different coccus of radiation hardness is connected on above-mentioned pGEMT-pprM recombinant plasmid, builds and has the recombinant plasmid pGEMT-pprMG of groEL containing complete pprM gene;
3. the recombinant plasmid pGEMT-pprMG importing pprM gene is proceeded in recipient E. coli JM109, obtain engineering strain JM_pprM;
Confirm through experiment, this bacterial strain has resists ability that 4M NaCl and high density (3M) sorbyl alcohol (Sorbitol) impact respectively (because Sorbitol has water absorbability, the Sorbitol solution of high density can the environment of simulating drought, detailed in Example 2);
4. D.radiodurans R1 pprM gene (DR_0907) is connected in plant expression vector pBI121, builds the recombinant plasmid pBI121-pprM with pprM gene;
5. being imported by pBI121-pprM recombinant plasmid does not have in the rape of drought resistance, and obtaining can the rapeseed plants of tolerate drought;
The transgene rape of the D.radiodurans R1 pprM gene that experimental result shows containing gained of the present invention obtains the stronger tolerance to arid (detailed in Example 5,6,7).
Accompanying drawing illustrates:
Fig. 1 is the PCR primer containing D.radiodurans R1 pprM gene (DR_0907) sequence;
Fig. 2 is the structure checking electrophoretogram of the prokaryotic expression carrier containing D.radiodurans R1 pprM gene (DR_0907) and groEL promotor;
Fig. 3 is the checking electrophoretogram of the plant expression vector construction containing D.radiodurans R1 pprM gene (DR_0907) sequence;
Fig. 4 is the colibacillary upgrowth situation containing empty carrier and the prokaryotic expression carrier containing D.radiodurans R1 pprM gene (DR_0907) sequence before shock test, wherein:
A is intestinal bacteria JM 109 bacterial strain containing empty expression vector;
B is the e. coli jm109 bacterial strain containing D.radiodurans R1 pprM gene (DR_0907) sequence expression vector;
Fig. 5 is intestinal bacteria (E.coli) upgrowth situation in containing the substratum of 3M sorbyl alcohol of prokaryotic expression carrier containing D.radiodurans R1 pprM gene (DR_0907) sequence and empty carrier, proves that D.radiodurans R1 pprM gene (DR_0907) can strengthen the function that intestinal bacteria have drought resisting.Bacterial strain in figure is as follows:
A is the e. coli jm109 bacterial strain containing empty expression vector;
B is the e. coli jm109 bacterial strain containing D.radiodurans R1 pprM gene (DR_0907) sequence expression vector.
Fig. 6 contains intestinal bacteria upgrowth situation in the substratum containing 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 salt tolerant resistance.Bacterial strain in figure is as follows:
A is the e. coli jm109 bacterial strain containing empty expression vector;
B is the e. coli jm109 bacterial strain containing D.radiodurans R1 pprM gene (DR_0907) sequence expression vector.
Fig. 7 is the RT-PCR checking of transgene rape.
Fig. 8 is the rape state before treatment turning pprM gene.
Fig. 9 is negative control (not genetically modified rape) state before treatment.
Figure 10 is the state that the rape turning pprM gene stops watering 1 week.
Figure 11 is the state that negative control (not genetically modified rape) stops watering 1 week.
Figure 12 is the state that the rape turning pprM gene stops watering 2 weeks.
Figure 13 is the state that negative control (not genetically modified rape) stops watering 2 weeks.
Figure 14 is transgene rape and wild-type rapeseed plant cell membranes in tissue permeability vs after drought stress, and wherein ordinate zou shows ion permeability.
Figure 15 is transgene rape and the contrast of wild-type rape leaf mda content after drought stress, and wherein ordinate zou shows mda content.
Embodiment
The plasmid lifted in following examples, bacterial strain, just for being described in further detail the present invention, are not limited flesh and blood of the present invention.All unreceipted specific experiment conditions, be according to normal condition well known to those skilled in the art, such as Sambrook equimolecular clone: laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) condition described in, or according to the condition that manufacturer advises.
The plasmid lifted in embodiment, bacterium source are as follows:
Cloning vector pGEMT-easy: be Promrga companies market product;
Plant expression vector pBI121: be Clontech companies market product;
Intestinal bacteria JM 109: be Beijing Quan Shi King Company commercially available prod.Agrobacterium EHA 105 is preserved by this laboratory
The expression of embodiment 1 D.radiodurans R1 pprM gene order in intestinal bacteria
Design a pair PCR Auele Specific Primer according to the pprM gene order in the D.radiodurans R1 genome announced, increase complete nucleotide sequence from D.radioduransR1 genomic dna.
Up 5′AGCGACTAGT GCCAAAT ATGTGCCGAGTTTC 3′；Down 5′ATCCCATATGTTACCAGCGGTCGTCGCGGC3′。From the genome of D.radiodurans R1, sequence SEQ ID NO:1 is amplified by PCR method, condition: 95 DEG C of 10min, [95 DEG C of 30sec, 55 DEG C of 30sec, 72 DEG C of 45sec] 30 circulations, 72 DEG C of 10min, PCR primer is after glue reclaims, be cloned on carrier pGEMT-easy, called after pGEMT-pprM, then the groEL promotor coming from D.radiodurans R1 is also linked on this carrier, build coli expression carrier pGEMT-pprMG, by this expression vector transformation of E. coli JM109, through PCR, enzyme is cut, the correct (see Fig. 1 of insertion sequences is verified in three aspects that check order, 2), be JM_pprM by this Strain Designation.
By the E.coli JM109 called after JM_pGEMT-G containing pGEMT-easy+groEL empty plasmid.
Embodiment 2 is containing the tolerance test of the engineering strain raising Drought and salt of D.radiodurans R1 pprM gene
One, experimental technique
1, the intestinal bacteria that two of gained in embodiment 1 are recombinated are inoculated in 20mL LB liquid nutrient medium, after shake-flask culture 3-4h, then transfer in the LB liquid nutrient medium of 100mL, keep the consistent of inoculum size as far as possible, be cultured between OD0.3-0.4, be adjusted to OD value consistent 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, aseptic deionized water proportional diluted to 10 used immediately by each sample
-4, get 10 μ L points at LB solid culture primary surface, cultivate 16h through 37 DEG C, observe and take a picture.The Sorbitol Powder of 4M NaCl salts solution and 3M is hypertonic solution, can Drought stress simulation.
Two, experimental result
As can be seen from Figure 4, the JM_pprM bacterial strain containing D.radiodurans R1 pprM gene (DR_0907) before impacting is basically identical with the JM_pGEMT-G strain growth state containing empty plasmid; After 4M NaCl salts solution and 3M sorbyl alcohol impact, JM_pprM strain growth containing D.radiodurans R1 pprM gene (DR_0907) in order, colony count is obviously more than the JM_pGEMT-G bacterial strain (see Fig. 4,5,6) only containing empty plasmid.
Three, conclusion
D.radiodurans R1 pprM gene can help coercing of prokaryotic organism tolerate drought and salt.
Embodiment 3 D.radiodurans R1 pprM gene carries out the qualification of eukaryotic cell expression and transfer-gen plant in rape cell
(1) containing the structure of goal gene plant expression vector
Be used for the primer of expression of plants according to D.radiodurans R1 genome sequence design pprM gene, upstream primer adds Bam HI, and downstream primer adds Sac I two restriction enzyme sites, 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 upstream is with Bam HI restriction enzyme site, downstream is with Sac I restriction enzyme site, pprM (DR_0907) gene is increased out 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 the expression vector pBI 121-pprM (see Fig. 3) containing complete DR0907 gene.
(2) agrobacterium mediation converted rape experiment
1. the competent preparation of agrobacterium tumefaciens EHA105:
1) picking list bacterium colony, be inoculated in 5mL YEB liquid nutrient medium (containing Rifampin Rif 50mg/L), 28 DEG C, 250rpm shaking culture is spent the night;
2) get 2mL bacterium liquid, add in 50mL YEB liquid nutrient medium (containing Rif 50mg/L), 28 DEG C, 250rpm shaking culture is to OD
600about about 0.6;
3) bacterium liquid is gone in 50mL sterile centrifugation tube, the centrifugal 5min of ice bath 30min, 5000 × g;
4) supernatant is abandoned, precipitation 2mL 20mM CaCl
2heavily floating, every part of 100 μ L are dispensed in 1.5mL centrifuge tube, save backup in liquid nitrogen.
2. recombinant plasmid dna proceeds to Agrobacterium:
1) the pBI-pprM plasmid DNA of about 1 μ g is joined respectively in 100 μ L EHA105 competent cells, mixing, ice bath 5min;
2) centrifuge tube is put freezing 8min in liquid nitrogen, go to rapidly temperature bath 5min in 37 DEG C of water-baths;
3) 1mL YEB liquid nutrient medium is added, 250rpm recovery 4 ~ 5h on 28 DEG C of shaking tables;
4) get on YEB solid medium that appropriate bacterium liquid is applied to containing Rifampin (Rif) 50mg/L and kantlex (Kan) 100mg/L, put 28 DEG C and cultivate 24 ~ 48h.
3. the extraction of the plasmid of Agrobacterium
1) picking colony in containing Rifampin (Rif) 50mg/L and kantlex (Kan) 100mg/L YEB liquid nutrient medium in (YEB: Tryptones 5g/L, yeast extract 1g/L, sucrose 5g/L, magnesium sulfate 0.5g/L), 28 DEG C, 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) wash 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, repeatedly aspirate mixing with pipettor, room temperature leaves standstill 10min;
3) add solution II (0.2M NaOH, 1%SDS) the 600 μ L of fresh configuration, turn upside down and mix several times;
4) add solution III (5M potassium acetate pH 5.2, Glacial acetic acid 11.5mL, adds water to 100mL) the 450 μ L of precooling, fully mix, ice bath 5min, 4 DEG C 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 (Tris-HCl 10mmol/L, 1mmol/LEDTA, pH 8.0), verify through PCR, BamHI, 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, the NaClO sterilizing 12min of 10%, again with 0.1% mercuric chloride solution sterilization 7-8min, with sterilizing washing 3-5 time, and on the Semen Brassicae campestris of sterilizing is placed in without hormone pre-culture medium equably (7.5% agar).Illumination cultivation, 4-5d Brassica Napus Seedling in age is most suitable for genetic transformation.
2) preculture
Cut cotyledon and vegetative point with the alcohol of 75% and the scissors of high-temp sterilizing, the hypocotyl that under clip rape aseptic seedling next-door neighbour vegetative point, about 0.5cm is long, be placed in pre-culture medium (MS+6-BA 2mg/L+2,4-D 1 mg/L+AgNO
32.5mg/L+AS 19.62mg/L) on, illumination preculture 2d.
3) cultivation of Agrobacterium
0.1mLpprM-EHA 105 bacterium liquid is added in 50mL LB liquid nutrient medium (containing kantlex 100mg/L, Rifampin 50mg/L), 220rpm about shaking culture 16h at 28 DEG C, the centrifugal 10min of 4000 × g under room temperature, abandon supernatant liquor, the MS liquid nutrient medium (containing AS 100 μm of ol/L) of thalline sterilizing suspends, be diluted to the 5-20 of original volume doubly, 220rpm shaking culture 1h at 28 DEG C, makes the OD of bacterium liquid
600reach about 0.5.
4) Dual culture
The hypocotyl of preculture 2d is put into bacterium liquid and contaminates 1min, hypocotyl pulled out by medication spoon, be placed on the thieving paper of sterilizing, blot the unnecessary bacterium liquid on hypocotyl, be put into again be coated with sterilizing filter paper pre-culture medium on, seal culture dish with sealed membrane, about 25 DEG C, be about 2d (light culture) in dark place Dual culture.
5) inducing culture
The hypocotyl of Dual culture 2d is put into inducing culture (MS+6-BA 2mg/L+AgNO
32.5mg/L), seal culture dish with sealed membrane, about 25 DEG C illumination cultivation, every two weeks subcultures once, until grow the callus expanded.
6) select to cultivate
The callus expanded is transferred to screening culture medium (MS+6-BA 2mg/L+AgNO
32.5mg/L+Kan 100mg/L) on, seal culture dish with sealed membrane, about 25 DEG C illumination cultivation, every two weeks subcultures once, until grow seedling.
7) root culture
When young shoot grows to 1-2cm height, they are separated from callus, transfer to (1/2MS+NAA 0.5mg/L+Kan 25mg/L) on root media and carry out root culture.Under antibiotic-screening condition, the bud of about 87% forms root after two weeks.
8) hardening and transplanting
When seedling after taking root grows to 5-6cm height, half opens wide culturing bottle lid, carries out hardening; After treating that seedling adapts to external environment, transfer to plantation in the vermiculite of indoor sterilizing and cultivate, and water with 1/2MS nutrient solution.When growth of seedling is to 7-9cm, transplants seedlings in earth and grow, and then carry out next step experiment.
Embodiment 4 utilizes RT-PCR method to detect the expression of pprM gene in transgene rape plant
The extraction of 1 plant total serum IgE
The post centrifugal a small amount of plant total serum IgE extraction agent box using Shanghai Hua Shun Bioisystech Co., Ltd to produce extracts the RNA turning pprM vector for transgenic rape obtained, and with DNase I process DNA.
2RT-PCR
Reverse transcription uses the test kit (DyNAmo of NEB company
tMcDNA Synthesis Kit).The cDNA obtained with reverse transcription is template, utilizes primer DR0907 PLANT UP:CGGCGGATCCATGTGCCGAGTTTCGATTGT;
DR0907 PLANT DOWN:ATTAGAGCTC TTACCAGCGGTCGTCGCGGC carries out PCR reaction, and condition is: 95 DEG C of 10min, [95 DEG C of 30sec, 55 DEG C of 30sec, 72 DEG C of 45sec] 30 circulation, 72 DEG C of 10min.
PCR primer through agarose gel electrophoresis detect after, sequence verification.Show that pprM gene has successfully been transferred in rape (see Fig. 7).
Drought resisting experiment (1) of embodiment 5 turns of pprM gene plants---rapeseed plant leaf water content measures
In intestinal bacteria, to drought stress, there is resistance in view of this nucleotide sequence has been proved to be, further Identification of Drought is carried out to transfer-gen plant.
To rape is disposable water sufficient nutritive medium after do not rewater, every other day measure leaf water content.The measuring method of water content: about 0.1g blade is from after plant takes off, take fresh weight (FW) immediately, then blade to be immersed in deionized water 4 hours, taking-up filter paper sucks surface-moisture, take saturated fresh weight (TW), then blade is put into baking oven to dry to constant weight in 70 DEG C, take dry weight (DW).
Relative water content (Relativewatercontent, RWC) (%)=(FW-DW)/(TW-DW) × 100%.
Plant is in Osmotic treatment process, and leaf r elative water content declines gradually, and along with the prolongation between arid place, the difference of transfer-gen plant relative water content and wild-type is obvious gradually.WT lines is when coercing the 4th day dry morning, and blade has started to wilt, and water content sharply declines; During genetically modified rape drought stress the 7th day, blade just starts to wilt, and leaves water loss speed is obviously slow than wild-type; After coercing 14 days dry morning, the leaf r elative water content of adjoining tree drops to 49%, and the relative water content of transgenic line is between 64-68%, far above adjoining tree (see Fig. 8,9,10,11,12,13).
Drought resisting experiment (2)---transgene rape and the wild-type rapeseed plant cell membranes in tissue permeability vs after drought stress of embodiment 6 turns of pprM gene plants
1) experiment purpose
When plant be subject to adverse circumstance affect time, after high temperature or low temperature, arid, salt marsh, pathogen infection, cytolemma is destroyed, and membrane permeability increases, thus makes intracellular Electrolyte Leakage, 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, the increase degree of more same crop different varieties membrane permeability at Different stress temperature, the resistance that can compare between crop is strong and weak.Therefore, conductivity meter method has become the cultivation of current crop anti-adversity, breeding has been identified one of the crop anti-adversity power accurately method of practicality.
Drought stress usually causes vegetable cell to damage, and causes cytolemma ion permeability to increase and lipid peroxdation.
2) experiment material
The rape that a turns the rape (preparation method is shown in embodiment 2) of pprM gene and wild-type simultaneously disposable water sufficient nutritive medium after do not rewater, the Osmotic treatment blade of the 8th day;
The transgenosis of b without Osmotic treatment and the rape leaf of wild-type;
3) experimental technique and step
Through the rape leaf of Osmotic treatment with not through the rape leave agreement that contracts a film or TV play to an actor or actress 0.1g of overdrying early process, 3 times are rushed with distilled water, blade surface moisture is blotted gently with thieving paper, blade is shredded and is placed in 25mL distilled water, in 0.05MPa lower pumping 20min in vacuum drier, slowly vibrate at 25 DEG C 2h, then measures specific conductivity S1, the blank specific conductivity C1 of Simultaneously test water.Again sample is processed 20min in boiling water bath, after being cooled to room temperature, constant volume measures specific conductivity S2 to 25mL, processes water in the same manner, measures its blank specific conductivity C2.Ion permeability=(S1-C1)/(S2-C2) × 100%.
4) experimental result
In untreated plant, ion permeability is between 12-15%; Osmotic treatment 8 days, the blade cell film ion permeability of the plant of wild-type reaches 32%; The ion permeability of transfer-gen plant is 22-24%, is significantly less than WT lines (see Figure 14).
Drought resisting experiment (3)---transgene rape and the comparison of wild-type rape leaf mda content after drought stress of embodiment 7 turns of pprM gene plants
1) experiment purpose
This experiment adopts thiobarbituricacidα-(TBA) colorimetry.Its principle is: plant is under adverse environmental factor, often peroxidation of membrane lipids occurs, mda (malondialdehyde, MDA) is one of its product, usual profit uses it as Lipid peroxidation index, represents the power that cell membrane lipid extent of peroxidation and plant react adverse environmental factor.
Mda reacts with thiobarbituricacidα-(TBA) under high temperature, acidic conditions, be formed in coloured trimethylammonium mixture that there is maximum light absorption at 532nm wavelength place, the specific absorbance of this mixture is 155 [mmol/ (L*cm)], and has minimum light to absorb at 600nm wavelength place.Can by following formula:
A
532-A
600＝155000×C×L (1)
Calculate MDA concentration C (μm ol/L), calculate MDA content C (μm ol/g) in the fresh tissue of unit weight further.A in formula
532and A
600represent the absorbance at 532nm and 600nm wavelength place respectively.L is cuvette thickness (cm).
It is pointed out that in plant tissue, glucide has interference effect to MDA-TBA reaction.For eliminating this interference, through test, available following formula eliminates the error caused by sucrose.
C/μmol/L＝6.45(A
532-A
600)-0.56A
450(2)
In formula: A
450, A
532, A
600represent the absorbance under 450nm, 532nm and 600nm wavelength respectively.Directly can try to achieve the concentration of MDA in plant sample extracting solution with formula (2), calculate its content in plant tissue further.
2) experiment material
The rape that a turns the rape (preparation method is shown in embodiment 2) of pprM gene and wild-type simultaneously disposable water sufficient nutritive medium after do not rewater, the Osmotic treatment blade of the 8th day;
The transgenosis of b without Osmotic treatment and the rape leaf of wild-type.
3) experimental technique and step
A gets 0.5g rape leaf, totally 4 parts, adds 5%TCA 5mL respectively, gained homogenate centrifugal 10min under 3000 × g after grinding;
B gets supernatant liquor 2mL, adds 0.67%TBA 2mL, boils 30min after mixing in 100 DEG C of water-baths, once centrifugal again after cooling;
C measures the absorbance of supernatant liquor under 450nm, 532nm and 600nm wavelength respectively, and calculates MDA concentration by formula (2), then calculates the MDA content (μm ol/g) in unit fresh weight tissue;
D the data obtained adopts the otherness of the different rape MDA content of variance analysis test two.
4) experimental result
After drought stress, the content of the mda of transfer-gen plant is also lower than wild-type (see Figure 15).
More than experiment proves that the resistance turning the rape of pprM gene in Deinococcus radiodurans R1 is far away higher than the rape of control group.
Claims (5)
1.Deinococcus radiodurans R1 pprM gene cultivates the purposes of drought-resistant plant, and the numbering of described gene and sequence number are DR_0907, GeneID:1797366.
2. the recombinant plasmid containing Deinococcus radiodurans R1 pprM gene is cultivating the purposes in drought-resistant plant.
3. purposes according to claim 2, described recombinant plasmid is can at the plasmid of escherichia coli expression.
4. purposes according to claim 2, described recombinant plasmid is the plasmid can expressed in plant.
5., containing the purposes of the host cell cultivation drought-resistant plant of the recombinant plasmid transformed of Deinococcus radiodurans R1 pprM gene, described host cell comprises prokaryotic cell prokaryocyte and eukaryotic cell.
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| Genome of the Extremely Radiation-Resistant Bacterium Deinococcus radiodurans Viewed from the Perspective of Comparative Genomics;KIRA S. MAKAROVA等;《MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS》;20010331 * |
| Harvey,D.等.GeneID:1797366.《GenBank》.2001, * |
| Identification of PprM: a modulator of the PprI-dependent DNA damage response in Deinococcus radiodurans;Hirofumi Ohba等;《Extremophiles》;20090226 * |
| 耐辐射球菌研究进展及其应用前景;舒海燕;《生态学杂志》;20091231 * |
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