CN106754497A - A kind of Ke Leishi pneumobacilluses and its microbial inoculum and preparation method and application - Google Patents
A kind of Ke Leishi pneumobacilluses and its microbial inoculum and preparation method and application Download PDFInfo
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- CN106754497A CN106754497A CN201611136507.5A CN201611136507A CN106754497A CN 106754497 A CN106754497 A CN 106754497A CN 201611136507 A CN201611136507 A CN 201611136507A CN 106754497 A CN106754497 A CN 106754497A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/22—Klebsiella
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/04—Pesticides, e.g. insecticides, herbicides, fungicides or nematocides
Abstract
The invention discloses a kind of Ke Leishi pneumobacilluses and its microbial inoculum and preparation method and application, described Ke Leishi pneumobacilluses (Klebsiella pneumoniae) FM84 be thiram degradation bacteria, depositary institution：China Committee for Culture Collection of Microorganisms's common micro-organisms center（CGMCC）, preservation day：On December 15th, 2015, deposit number：CGMCC No. 11663；The Ke Leishi pneumobacilluses belong to bacterium circle, Proteobacteria, γ deformation Gammaproteobacterias, enterobacteria mesh, enterobacteriaceae, Klebsiella.Described Ke Leishi pneumobacilluses for active component microbial inoculum be with the Ke Leishi pneumobacilluses (Klebsiella pneumoniae) FM84 prepared through actication of culture, seed culture, productive culture, preparation after the microbial inoculum that is made.Preparation method includes activation culture and fermented and cultured step；Application of the described microbial inoculum in degraded thiram agriculture thing is prepared.Microbial inoculum preparation process is simple containing the bacterial strain, it is with low cost, it is easy to use, with good application prospect.
Description
Technical field
The invention belongs to microbial technology field, and in particular to a kind of Ke Leishi pneumobacilluses and its microbial inoculum and preparation method
With application.
Background technology
Thiram is the common pesticides in plant mycosis disease control, and chemical name is the thio carbon of peroxidating two of tetramethyl
Sour diamides, and commonly referred to as thiuram, TMTD etc..English name：tetramethylthiuram disulfide；
Thiram.White, has special odor, crystalline powder at canescence.Density is slightly larger than water.It is water insoluble, insoluble in sig water, vapour
Oil, is dissolved in ethanol, benzene, chloroform, carbon disulfide etc..Thiram and Mancozeb pass through frequently as two kinds of active ingredients multiple to be fabricated to
Distribution type bactericide, belongs to low toxicity bactericide, has stimulation to skin and mucous membrane.The fungus diseases such as epidemic disease can be treated, extensively
As bactericide, downy mildew, epidemic disease, anthracnose to various crop, Cereal smut, seedling stage yellow blight have preferably
Preventing and treating, thiram is conventional fungal disease pesticide control on tobacco.Due to applying in large quantities for a long time, thiram in tobacco leaf
Recall rate more and more higher.Because thiram and its derivative have carcinogenicity, mutagenicity and a teratogenesis, and acute toxicity and
Sub-lethal effect, has caused the very big concern of people at present.Biodegradation be to thiram pollution tobacco leaf degraded it is important
One of approach.For thiram residual, the screening operation of thiram degrading microorganism has been carried out both at home and abroad.Up to the present,
Multiple bacterial strains with thiram degradation capability, such as pseudomonas, Mycobacterium, bacillus are isolated, but
Have no pneumobacillus for the report in thiram degraded.
The content of the invention
An object of the present disclosure is to provide a kind of Ke Leishi pneumobacilluses, and the second purpose is to provide one kind with described gram
Lei Shi pneumobacilluses as active component microbial inoculum, the 3rd purpose be provide it is described using Ke Leishi pneumobacilluses as activity into
The bacterial preparation process for dividing, the 4th purpose is to provide the answering using Ke Leishi pneumobacilluses as the microbial inoculum of active component
With.
The first object of the present invention be achieved in that described Ke Leishi pneumobacilluses (Klebsiella pneumoniae)
FM84 is thiram degradation bacteria, depositary institution：China Committee for Culture Collection of Microorganisms's common micro-organisms center（CGMCC）,
Preservation day：On December 15th, 2015, deposit number：CGMCC No. 11663；The Ke Leishi pneumobacilluses belong to bacterium circle,
Proteobacteria, γ-deformation Gammaproteobacteria, enterobacteria mesh, enterobacteriaceae, Klebsiella.
--- Ke Leishi pneumobacilluses (Klebsiella pneumoniae) FM84 acquisition with identification
（1）Ke Leishi pneumobacilluses (Klebsiella pneumoniae) FM84 acquisition with identification
Ke Leishi pneumobacilluses of the present invention, are isolated from tobacco leaf.
Bacterial strain is obtained：
LB fluid nutrient mediums：Dusty yeast 10g, peptone 5.0g, sodium chloride 10.0g, distilled water complements to 1000ml, is stirred after mixing
Mix uniform, natural ph, high pressure steam sterilization (121 DEG C, 30min) is obtained afterwards.
LB solid mediums：Agar 15.0g is added in LB Liquid Culture based formulas.
Tobacco sample picks up from Yuxi Hongta District of Yunnan Province, takes 1g tobacco samples and is placed in 250ml conical flasks, adds
The thiram of 100ml LB fluid nutrient mediums and final concentration of 100mg/L, shaken cultivation (28 DEG C, 150rpm) 5 days, takes nutrient solution
Gradient dilution is carried out, the μ l of nutrient solution 200 after dilution is taken and is coated on the LB solid plates containing 100mg/L thiram, be placed in perseverance
Culture in warm incubator (28 DEG C), after bacterium colony is grown on flat board, each bacterium colony of picking is flat in the LB solids containing 100mg/L thiram
Purified repeatedly on plate, until bacterium colony is single, each bacterium colony after purification is respectively connected to the LB liquid of final concentration of 100mg/L thiram
Shaken cultivation (28 DEG C, 150rpm) overnight, the centrifugation of cultured bacterium solution is followed by cultivating one into enrichment culture liquid in body test tube
In week, thiram residual quantity in each enrichment culture liquid is detected by high performance liquid chromatography (HPLC), finally screening obtains one plant of energy
The bacterial strain of degraded effective cypermethrin, is named as FM84.
Identification of strains：
The bacterial strain of above-mentioned acquisition is carried out into biochemical character and molecular biology identification.The main biological property of the bacterial strain is：Bacterium
Body is without gemma and flagellum, and Gram-negative is milky on LB solid mediums, opaque, there is a pod membrane, viscous toughness,
Subcircular, neat in edge.Results of biochemical is shown in Table 1.Bacterial strain can be at 25~35 DEG C, under the condition of culture of pH value 7.0~9.0 preferably
Growth.The bacterial strain is accredited as the Ke Leishi pneumobacilluses of pneumobacillus category through 16S rDNA sequence analyses（Klebsiella pneumoniae）.
The FM84 bacterial strains biochemical character of table 1 is determined
In table 1, "+" represents positive reaction, and "-" represents negative reaction.
（2）Ke Leishi pneumobacilluses (Klebsiella pneumoniae) FM84 preservation
By above-mentioned qualification result, confirm bacterial strain FM84 be Ke Leishi pneumobacilluses (Klebsiella pneumoniae) one
Individual strain, is named as FM84, and the common micro- life of China Committee for Culture Collection of Microorganisms is preserved on December 15th, 2015
Thing center（Abbreviation CGMCC, address is：Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode
100080）, its deposit number：CGMCC No. 11663.
The second object of the present invention be achieved in that with the Ke Leishi pneumobacilluses (Klebsiella pneumoniae) FM84 prepared through actication of culture, seed culture, productive culture, preparation after the microbial inoculum that is made.
The third object of the present invention is achieved in that described Ke Leishi pneumobacilluses are the system of the microbial inoculum of active component
Preparation Method, it is characterised in that including actication of culture, seed culture, productive culture, preparation preparation process, specifically include：
(1) actication of culture：The preservation kind of the FM84 bacterial strains is cultivated by plating method, culture to single bacterium colony occurs；
(2) seed culture：By the single bacterium colony access to seed culture bottle in, with FM84 bacterial strain seed culture medium cultures to logarithm
Growth period；
(3) productive culture：In the nutrient solution access fermentation tank that step (2) culture is obtained, trained with FM84 bacterial strains production medium
Support to exponential phase, the access amount of the nutrient solution is the 4-8% of the FM84 bacterial strains production medium cumulative volume；
(4) prepared by preparation：The zymotic fluid centrifugation that step (3) culture is obtained, precipitation is suspended with physiological saline, and packing obtains Fu Mei
Double bacterium for degradating residual agricultural chemical.
The fourth object of the present invention is achieved in that application of the described microbial inoculum in degraded thiram medicine is prepared.
Ke Leishi pneumobacilluses of the invention (Klebsiella pneumoniae) FM84, low production cost, thiram agriculture
Medicine removal effect is good, efficient, nontoxic, environmental protection, and the bacterial strain can be prepared by simple, quick, inexpensive operating method
Thiram bacterium for degradating residual agricultural chemical, the degradation bacterial agent prepared using bacterial strain of the present invention has low production cost, user
Just, the advantages of removal effect is good, can be effective for the production of non agricultural chemical residuum agricultural product in agricultural production.
Outer unless otherwise indicated, the percentage employed in the present invention is percentage by volume.
Brief description of the drawings
Fig. 1 be Ke Leishi pneumobacilluses of the present invention (Klebsiella pneumoniae) FM84 LB culture mediums bacterium colony
Feature schematic diagram；
Fig. 2 be Ke Leishi pneumobacilluses of the present invention (Klebsiella pneumoniae) FM84 shows the degradation curve of thiram
It is intended to.
Specific embodiment
With reference to embodiment and accompanying drawing, the present invention is further illustrated, but the present invention is subject to never in any form
Limitation, based on present invention teach that any conversion or improvement made, each fall within protection scope of the present invention.
Ke Leishi pneumobacilluses of the present invention (Klebsiella pneumoniae) FM84 be thiram degradation bacteria,
Depositary institution：China Committee for Culture Collection of Microorganisms's common micro-organisms center（CGMCC）, preservation day：In December, 2015
15 days, deposit number：CGMCC No. 11663；The Ke Leishi pneumobacilluses belong to bacterium circle, Proteobacteria, γ-mycetozoan
Guiding principle, enterobacteria mesh, enterobacteriaceae, Klebsiella.
Described Ke Leishi pneumobacilluses (Klebsiella pneumoniae) biological property of FM84 is：Thalline without
Gemma and flagellum, Gram-negative is milky on LB solid mediums, opaque, there is pod membrane, viscous toughness, nearly circle
Shape, neat in edge.Bacterial strain can preferably grow at 25~35 DEG C under the condition of culture of pH value 7.0~9.0.
Ke Leishi pneumobacilluses of the present invention are the microbial inoculum of active component, are with the Ke Leishi pneumobacilluses
(Klebsiellapneumoniae) FM84 prepared through actication of culture, seed culture, productive culture, preparation after the microbial inoculum that is made.
Ke Leishi pneumobacilluses of the present invention are the preparation method of the microbial inoculum of active component, it is characterised in that including bacterium
Activation, seed culture, productive culture, preparation preparation process are planted, is specifically included：
Actication of culture：The preservation kind of the FM84 bacterial strains is cultivated by plating method, culture to single bacterium colony occurs；Seed
Culture：By the single bacterium colony access to seed culture bottle in, with FM84 bacterial strain seed culture medium cultures to exponential phase；Production
Culture：The nutrient solution that step seed culture is obtained is accessed in fermentation tank, is given birth to FM84 bacterial strain production medium cultures to logarithm
For a long time, the access amount of the nutrient solution is the 4-8% of the FM84 bacterial strains production medium cumulative volume；Seed culture and production are trained
The optimization formula for supporting base contains：(NH4)2SO4 1-3g/L、K2HPO4 4-7g/L、KH2PO41-3g/L、MgSO4•7H2O 0.1g/L、
Glucose 5-10 g/L.Condition of culture is：25~35 DEG C of cultivation temperature, pH value 7.0~9.0,150~200rpm of stirring frequency,
Air mass flow is 0.1 ~ 1.0vvm.It is prepared by preparation：The zymotic fluid centrifugation that step productive culture is obtained（6000rpm, 10min）,
Precipitate and suspended with the sodium-chloride water solution containing 0.5% Tween-20 and 0.9%, packing obtains thiram pesticide residual degrading bacteria
Agent.
Ke Leishi pneumobacilluses of the present invention are that described microbial inoculum is preparing poison for the application of the microbial inoculum of active component
Application in degraded thiram medicine.
With specific embodiment, the present invention will be further described below：
Embodiment 1
The screening of bacterial strain and identification
Leaf sample is gathered from the tobacco of grown in field, nature enrichment culture is carried out as bacterium source, by glucose 2g, tryptose
Peptone 4g, beef extract 4g, MgSO40.8g is added in the distilled water of 400mL, temperature conditionss of the nutrient solution for preparing at 121 DEG C
Lower sterilization treatment 20min, cultivates three days on this culture medium, then prepares inorganic salts basal medium, adds thiram, is configured to
After solution, then add distilled water and be settled to 500mL, good fortune is beautiful during the amount of above-mentioned addition thiram will make the nutrient solution for preparing
Double ultimate densities is 100mg/mL, the nutrient solution that then will be prepared sterilization treatment under 121 DEG C of temperature conditionss
20min, obtains final product required nutrient solution.Cultivated three days on this culture medium, dilution spread isolates one plant of bacterium and is named as FM84, should
Bacterium was preserved in " China Committee for Culture Collection of Microorganisms's common micro-organisms center " on December 15th, 2015, its preserving number
CGMCC 11663。
The PCR amplifications of the 16SrDNA genes of FM84, sequencing are completed by Shanghai Ying Jun Bioisystech Co., Ltd.Bacterial strain
FM84 16S rDNA sequences（GenBank registration numbers：KU057959）It is nearest with Klebsiella bacterium affiliation, withKlebsiella pneumoniaeCP010361.1 bacterial strains homology up to 100.0%, give birth to by combining form, colony characteristicses and physiology
Change characteristic, identification bacterial strain belongs to K. pneumonia（Klebsiella pneumoniae）.
Embodiment 2
The preparation of microbial inoculum
The preservation kind of the FM84 bacterial strains is inoculated into LB solid plates cultivation carries out culture 24h, occurs to single bacterium colony；Will be upper
State single bacterium colony and access to and contain (NH4) 2SO4 1g/L, K2HPO4 4g/L, KH2PO4 3g/L、MgSO4•7H2O 0.1g/L, Portugal
In the triangular flask of grape 5 g/L culture mediums of sugar, culture to exponential phase obtains seed liquor；The nutrient solution that seed culture is obtained is connect
Enter in fermentation tank, inoculative proportion is 5%, and 28 DEG C, 200rpm, air mass flow is 0.8vvm, culture 48h obtains productive culture liquid.Will
The zymotic fluid centrifugation that productive culture is obtained（6000rpm, 10min）, sodium chloride of the precipitation containing 0.5% Tween-20 and 0.9%
The aqueous solution suspends, and packing obtains thiram bacterium for degradating residual agricultural chemical.
Embodiment 3
Thiram degradation experiment
The crop field tobacco leaf polluted by thiram, 1-10 × 10 are sprayed by every mu13Cfu K. pneumonias（Klebsiella pneumoniae）FM84, cigarette strain can realize the biodegradation to thiram residual on tobacco leaf after field grows 7d naturally.
Fig. 2 illustrate FM84 bacterium treatment tobacco leaf in thiram with the time dynamics degradation curve.Inoculation thiram degraded
The treatment of bacterium FM84 thiram when 7 days degrades 99.2%, and spray clear water is slow as the CK tobacco leaves degraded of control, and 7
It when degrade 67.2%.It can be seen that after thiram degradation bacteria is sprayed, accelerating the degraded of thiram residual.
It is good to the biodegradable effect that thiram on tobacco leaf is remained using the method for the present invention, with thiram natural degradation phase
Than, the degradation effect that tobacco leaf is polluted to thiram is improve, it is more beneficial for the control in practice to thiram residues of pesticides on tobacco leaf
System.
Claims (9)
1. a kind of Ke Leishi pneumobacilluses, it is characterised in that described Ke Leishi pneumobacilluses (Klebsiella pneumoniae)
FM84 is thiram degradation bacteria, depositary institution：China Committee for Culture Collection of Microorganisms's common micro-organisms center
（CGMCC）, preservation day：On December 15th, 2015, deposit number：CGMCC No. 11663；The Ke Leishi pneumobacilluses belong to
Bacterium circle, Proteobacteria, γ-deformation Gammaproteobacteria, enterobacteria mesh, enterobacteriaceae, Klebsiella.
2. Ke Leishi pneumobacilluses according to claim 1, it is characterised in that described Ke Leishi pneumobacilluses
(Klebsiella pneumoniae) biological property of FM84 is：Thalline is without gemma and flagellum, and Gram-negative is solid in LB
It is milky on body culture medium, it is opaque, there are pod membrane, viscous toughness, subcircular, neat in edge；Bacterial strain can at 25~35 DEG C,
Preferably grown under the condition of culture of pH value 7.0~9.0.
3. the Ke Leishi pneumobacilluses described in a kind of claim 1 or 2 are the microbial inoculum of active component, it is characterised in that be with described
Ke Leishi pneumobacilluses (Klebsiella pneumoniae) FM84 is through actication of culture, seed culture, productive culture, preparation system
The microbial inoculum being made after standby.
4. the Ke Leishi pneumobacilluses described in a kind of claim 3 are the preparation method of the microbial inoculum of active component, it is characterised in that
Including through actication of culture, seed culture, productive culture, preparation preparation process, specifically including：
(1) actication of culture：The preservation kind of the FM84 bacterial strains is cultivated by plating method, culture to single bacterium colony occurs；
(2) seed culture：By the single bacterium colony access to seed culture bottle in, with FM84 bacterial strain seed culture medium cultures to logarithm
Growth period；
(3) productive culture：In the nutrient solution access fermentation tank that step (2) culture is obtained, trained with FM84 bacterial strains production medium
Support to exponential phase, the access amount of the nutrient solution is the 4-8% of the FM84 bacterial strains production medium cumulative volume；
(4) prepared by preparation：The zymotic fluid centrifugation that step (3) culture is obtained, precipitation is suspended with physiological saline, and packing obtains Fu Mei
Double bacterium for degradating residual agricultural chemical.
5. Ke Leishi pneumobacilluses according to claim 4 are the preparation method of the microbial inoculum of active component, it is characterised in that
Described seed culture and the optimization formula of production medium contain：(NH4)2SO4 1-3g/L、K2HPO4 4-7g/L、KH2PO41-
3g/L、MgSO4•7H2O 0.1g/L, glucose 5-10 g/L.
6. Ke Leishi pneumobacilluses according to claim 4 are the preparation method of the microbial inoculum of active component, it is characterised in that
Condition of culture in seed culture and productive culture is：25~35 DEG C of cultivation temperature, pH value 7.0~9.0, stirring frequency 150~
200rpm, air mass flow is 0.1 ~ 1.0vvm.
7. Ke Leishi pneumobacilluses according to claim 4 are the preparation method of the microbial inoculum of active component, it is characterised in that
Zymotic fluid centrifugal condition in prepared by preparation is 6000rpm, and 10min, physiological saline is to contain 0.5% Tween-20 and 0.9%
Sodium-chloride water solution.
8. Ke Leishi pneumobacilluses according to claim 4 are the preparation method of the microbial inoculum of active component, it is characterised in that
FM84 thalline content is more than 1 × 10 in degradation bacterial agent10Individual/mL.
9. the Ke Leishi pneumobacilluses described in a kind of claim 3 are the application of the microbial inoculum of active component, it is characterised in that described
Microbial inoculum degrade thiram agricultural chemicals in application.
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Cited By (4)
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| CN108277185A (en) * | 2018-04-09 | 2018-07-13 | 中国林业科学研究院亚热带林业研究所 | A kind of turpentine oil microbial inoculum for degrading |
| CN111593000A (en) * | 2020-05-18 | 2020-08-28 | 西北农林科技大学 | Plastic mulching film degrading bacteria and application thereof |
| CN114561323A (en) * | 2022-02-23 | 2022-05-31 | 浙江台州秀川科技有限公司 | Salt-tolerant Klebsiella pneumoniae capable of efficiently degrading gallic acid and application thereof |
| CN115895937A (en) * | 2022-08-23 | 2023-04-04 | 华南农业大学 | Klebsiella pneumoniae obtained through microgravity mutagenesis breeding and culture method and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108277185A (en) * | 2018-04-09 | 2018-07-13 | 中国林业科学研究院亚热带林业研究所 | A kind of turpentine oil microbial inoculum for degrading |
| CN111593000A (en) * | 2020-05-18 | 2020-08-28 | 西北农林科技大学 | Plastic mulching film degrading bacteria and application thereof |
| CN111593000B (en) * | 2020-05-18 | 2022-07-22 | 西北农林科技大学 | Klebsiella for degrading plastic mulching film and application thereof |
| CN114561323A (en) * | 2022-02-23 | 2022-05-31 | 浙江台州秀川科技有限公司 | Salt-tolerant Klebsiella pneumoniae capable of efficiently degrading gallic acid and application thereof |
| CN114561323B (en) * | 2022-02-23 | 2024-04-09 | 浙江台州秀川科技有限公司 | Klebsiella pneumoniae capable of resisting salt and efficiently degrading gallic acid and application thereof |
| CN115895937A (en) * | 2022-08-23 | 2023-04-04 | 华南农业大学 | Klebsiella pneumoniae obtained through microgravity mutagenesis breeding and culture method and application thereof |
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