CN110628626A - Method for separating tobacco mosaic virus resistant bacteria from soil - Google Patents
Method for separating tobacco mosaic virus resistant bacteria from soil Download PDFInfo
- Publication number
- CN110628626A CN110628626A CN201910981804.7A CN201910981804A CN110628626A CN 110628626 A CN110628626 A CN 110628626A CN 201910981804 A CN201910981804 A CN 201910981804A CN 110628626 A CN110628626 A CN 110628626A
- Authority
- CN
- China
- Prior art keywords
- mosaic virus
- tobacco mosaic
- taking
- soil
- tobacco
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- 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
- C12N1/02—Separating microorganisms from their culture media
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/18—Testing for antimicrobial activity of a material
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Toxicology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for separating tobacco mosaic virus resisting bacteria from soil, which comprises the following steps: (1) taking 1.0g of soil sample, placing into a bottle, adding sterile water, sealing, and culturing for 1-3h with shaking; (2) standing the solution subjected to shake culture in the step (1) for 20-60min, taking supernatant for gradient dilution, uniformly mixing, and respectively uniformly coating 80-120 mu L of each concentration on a flat plate of a culture medium; (3) culturing the flat plate in a constant temperature incubator at 25-30 ℃ for 15-20 h; (4) and taking out the cultured plate, selecting a plate with uniformly distributed bacterial colonies from the plate, picking a single spot, placing the single spot into 2-5mL of culture solution, and performing shake culture for 10-20h to form a bacterial solution. The invention discloses a method for separating bacteria with inhibiting effect on tobacco mosaic virus from samples collected from different habitats (shallow sea mud and tobacco rhizosphere soil), enriching the types of beneficial bacteria with antiviral effect, further simply determining the antiviral effect and laying a foundation for the utilization of the bacteria with the antiviral effect.
Description
Technical Field
The invention relates to the technical field of microbial ecology, in particular to a method for separating tobacco mosaic virus resistant bacteria from soil.
Background
Tobacco Mosaic Virus (TMV) is a pathogen of tobacco mosaic disease and the like, has a wide host range, can infect at least 125 plants of 9 families, such as cruciferae, solanaceae, cucurbitaceae and the like, has high occurrence frequency, is quick in prevalence, is serious in harm and difficult to treat, cannot be effectively controlled once a plant is infected, is very easy to cause the prevalence of the virus disease, and causes serious economic loss. The economic losses due to tobacco mosaic virus are as high as billions of dollars per year. At present, modes such as breeding of antiviral varieties, control of virus-carrying media, cross protection and the like are mainly adopted in the treatment of virus diseases, and few effective chemical pesticides aiming at plant virus diseases need to be solved by agricultural scientific researchers.
It has been studied that plant viruses lose their infectivity rapidly after they become contaminated with bacteria, and some researchers have been working on the antiviral activity of microorganisms and their metabolites. At present, many bacteria, fungi and metabolites thereof have certain capability of inactivating tobacco mosaic virus particles, but no good method exists for separating the tobacco mosaic virus resisting bacteria.
Therefore, it is necessary to provide a further solution to the above problems.
Disclosure of Invention
The invention aims to provide a method for separating tobacco mosaic virus resistant bacteria from soil so as to overcome the defects in the prior art.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a method for isolating tobacco mosaic virus resistant bacteria from soil comprising the steps of:
(1) taking 1.0g of soil sample, placing into a bottle, adding sterile water, sealing, and culturing for 1-3h with shaking;
(2) standing the solution subjected to shake culture in the step (1) for 20-60min, taking supernatant for gradient dilution, uniformly mixing, and respectively uniformly coating 80-120 mu L of each concentration on a flat plate of a culture medium;
(3) culturing the flat plate in a constant temperature incubator at 25-30 ℃ for 15-20 h;
(4) taking out the cultured plate, selecting a plate with uniformly distributed bacterial colonies from the plate, picking out a single spot, placing the single spot into 2-5mL of culture solution, and performing shake culture for 10-20h to form a bacterial solution;
(5) centrifuging 1mL of bacterial liquid for 8-12min, mixing the supernatant with the isovolumetric tobacco mosaic virus inoculation liquid, and standing at room temperature for 15-30min to form a mixed liquid;
(6) mixing tobacco mosaic virus inoculation liquid with an equal volume of phosphate buffer solution, and standing at room temperature for 15-30min to form a control mixed liquid;
(7) uniformly spreading quartz sand on the Sansheng tobacco leaves by taking the Sansheng tobacco as an experimental material, and slightly rubbing the mixed liquid on half leaves by taking the main veins of the Sansheng tobacco leaves as a boundary; the other half of the leaves are rubbed to contrast the mixed solution;
(8) rubbing the mixed solution and the control mixed solution for 15-30min, spraying sterile water to clean the leaves, and naturally air-drying at room temperature;
(9) carrying out isolation culture in a greenhouse for 2-4d, then counting the number of dead spots on leaves, carrying out one-factor variance analysis on experimental data, carrying out significant difference analysis, and counting the antiviral effect according to the following formula:
preferably, 7-12mL of sterile water is added in step (1).
Preferably, the step (1) and/or the step (4) is/are oscillated at 100-140rpm at 25-30 ℃.
Preferably, the supernatant liquid obtained in the step (2) is diluted by a gradient 100、10-1、10-2、10-3、10-4And 10-5。
Preferably, the culture medium adopts LB solid culture medium.
Preferably, the culture solution is an LB liquid medium.
Preferably, the soil sample is taken from shallow sea mud or tobacco rhizosphere soil.
Preferably, the three-generation tobacco is cultured under the conditions of illumination time of 16h, illumination intensity of 2000lux, dark time of 8h, temperature of 25 +/-1 ℃ and relative humidity of 60%.
Preferably, step (7) employs three-generation tobacco cultured for four weeks.
Preferably, the preparation method of the tobacco mosaic virus inoculation liquid comprises the following steps:
(1) taking part of tobacco leaves infected with tobacco mosaic virus, grinding with 10mmol/L, pH 7.4.4 phosphate buffer solution, and homogenizing;
(2) filtering the serous fluid generated in the step (1) by using double-layer gauze, and centrifuging;
(3) taking the supernatant centrifuged in the step (2), treating the supernatant with polyethylene glycol twice, and then centrifuging again;
(4) and (4) after the final centrifugation in the step (3), taking the precipitate, and carrying out heavy suspension by using 10mmol/L, pH 7.4.4 phosphate buffer solution to obtain the tobacco mosaic virus inoculation solution.
Preferably, the preparation of the tobacco mosaic virus inoculum is operated at 4 ℃.
Compared with the prior art, the invention has the beneficial effects that:
the invention discloses a method for separating tobacco mosaic virus resistant bacteria from soil, which separates bacteria with inhibiting effect on tobacco mosaic virus from samples collected from different habitats (shallow sea mud and tobacco rhizosphere soil), enriches the types of beneficial bacteria for resisting virus, further simply determines the antiviral effect, and lays a foundation for the utilization of the tobacco mosaic virus resistant bacteria.
Detailed Description
The disclosure may be understood more readily by reference to the following detailed description of preferred embodiments of the invention and the examples included therein. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control. The term "prepared from …" as used herein is synonymous with "comprising". The terms "comprises," "comprising," "includes," "including," "has," "having," "contains," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
The conjunction "consisting of …" excludes any unspecified elements, steps or components. If used in a claim, the phrase is intended to claim as closed, meaning that it does not contain materials other than those described, except for the conventional impurities associated therewith. When the phrase "consisting of …" appears in a clause of the subject matter of the claims rather than immediately after the subject matter, it defines only the elements described in the clause; other elements are not excluded from the claims as a whole.
When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted to include the ranges "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a range of values is described herein, unless otherwise stated, the range is intended to include the endpoints thereof and all integers and fractions within the range.
The singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. "optional" or "any" means that the subsequently described event or events may or may not occur, and that the description includes instances where the event occurs and instances where it does not.
Approximating language, as used herein throughout the specification and claims, is intended to modify a quantity, such that the invention is not limited to the specific quantity, but includes portions that are literally received for modification without substantial change in the basic function to which the invention is related. Accordingly, the use of "about" to modify a numerical value means that the invention is not limited to the precise value. In some instances, the approximating language may correspond to the precision of an instrument for measuring the value. In the present description and claims, range limitations may be combined and/or interchanged, including all sub-ranges contained therein if not otherwise stated.
In addition, the indefinite articles "a" and "an" preceding an element or component of the invention are not intended to limit the number requirement (i.e., the number of occurrences) of the element or component. Thus, "a" or "an" should be read to include one or at least one, and the singular form of an element or component also includes the plural unless the stated number clearly indicates that the singular form is intended.
Example 1:
the plant material used in the examples of the present application was a tobacco mosaic virus, a blight host, a third generation tobacco (nicotiana. tabacum cv. sunnamann), which was bred by the present applicant, and cultivated in a greenhouse. Plant culture conditions: the illumination is 16h, the temperature is 25 +/-1 ℃, the illumination intensity is 2000lux, and the relative humidity is 60 percent; the dark time is 8h, the temperature is 25 +/-1 ℃, and the relative humidity is 60%. The nutrient medium is produced by a Ward nutrient soil processing factory in Shouguang city in Shandong province, and additional fertilization is not needed during the growth of the plants.
The tobacco mosaic virus source is collected from Qingdao city, Shandong province, and propagated and stored on common tobacco NC89 cultivated in a greenhouse.
The soil samples are respectively collected from tobacco rhizosphere soil and shallow sea mud.
Preparing tobacco mosaic virus inoculation liquid: the tobacco mosaic virus solution was prepared using the method of Gooding & Helbert (1967) with minor modifications. Taking new upper leaves of NC89 infected with tobacco mosaic virus, grinding with 10mmol/L phosphate buffer solution (pH 7.4), and homogenizing; then filtered with double layer gauze and centrifuged (1000 Xg, 20 min); the supernatant was taken and treated twice with polyethylene glycol. Centrifuging again (10000 Xg, 30 min); the precipitate was resuspended in 10mmol/L PBS buffer (pH 7.4) to obtain tobacco mosaic virus inoculum. The whole operation is carried out at 4 ℃, then a spectrophotometer is used for measuring the absorbance value of 260nm wavelength, and the virus concentration is calculated according to a formula.
Concentration of virus (mg/mL) ═ A260 Xdilution factor/E0.1％ 1cm260nm
Note: e is the extinction coefficient, i.e. the value of the optical absorption (optical density) at an optical path length of 1crn for a suspension with a concentration of 0.1% (1mg/mL) at a wavelength of 260 nm. Tobacco mosaic Virus E0.1％ 1cmThe concentration of the virus mother liquor is 10 mug/mL, and the concentration of the virus mother liquor is 3.1 at 260 nm.
A method for isolating tobacco mosaic virus resistant bacteria from soil comprising the steps of:
(1) a1.0 g soil sample was taken, placed in a sterile Erlenmeyer flask, added with 9mL of sterile water (operating in a clean bench), sealed and cultured with shaking for 2 hours (28 ℃,120 rpm).
(2) Taking out, standing in a super clean bench for 30min, and taking supernatant for gradient dilution 100、10-1、10-2、10-3、10-4And 10-5After mixing, 100. mu.L of each concentration was applied to a plate of LB solid medium.
(3) The LB plates were incubated for 16h at 28 ℃ in an incubator.
(4) Taking out, selecting a plate with uniformly distributed bacterial colonies (with moderate colony number), picking a single spot, and placing the single spot into 3mL of LB liquid culture medium for shake culture for 16h (28 ℃,120 rpm).
(5) Taking 1mL of bacterial liquid, centrifuging at 6000 Xg for 10min, taking supernatant, mixing with the tobacco mosaic virus inoculation liquid with the same volume, and standing at room temperature for 20 min.
(6) Mixing tobacco mosaic virus inoculation liquid with phosphate buffer solution of the same volume, and standing at room temperature for 20min to form a control mixed liquid;
(7) the antiviral effect of the bacterial liquid is detected by adopting a 'half-leaf method': uniformly spreading quartz sand on Sansheng tobacco leaves by taking Sansheng tobacco (NN) cultured for 4 weeks as an experimental material, dipping the mixed solution by using a sterile cotton swab, and slightly rubbing half leaves by taking a main vein as a boundary; the other half of the leaves are rubbed and controlled to form seed mixture;
(8) rubbing for 20min, spraying sterile water to clean the leaves, and naturally air drying at room temperature;
(9) isolating (far away from healthy tobacco) in a greenhouse and culturing for 3d, then counting the number of dead spots on leaves, carrying out single-factor variance analysis on experimental data by using an SPSS v18.0 to carry out Duncan new double-range method, carrying out significant difference analysis, and counting the antiviral effect according to the following formula:
example 2:
(1) a1.0 g soil sample was taken, placed in a sterile Erlenmeyer flask, added with 11mL of sterile water (operating in a clean bench), sealed and cultured with shaking for 3 hours (26 ℃,130 rpm).
(2) Taking out, standing in a super clean bench for 50min, and taking supernatant for gradient dilution 100、10-1、10-2、10-3、10-4And 10-5After mixing, 110. mu.L of each concentration was applied to a plate of LB solid medium.
(3) LB plates were incubated for 20h at 26 ℃ in an incubator.
(4) Taking out, selecting a plate with uniformly distributed bacterial colonies (with moderate colony number), picking a single spot, and placing the single spot into 4mL of LB liquid culture medium for shake culture for 20h (26 ℃,130 rpm).
(5) Taking 1mL of bacterial liquid, centrifuging at 6000 Xg for 10min, taking supernatant, mixing with the tobacco mosaic virus inoculation liquid with the same volume, and standing at room temperature for 25 min.
(6) Mixing tobacco mosaic virus inoculation liquid with phosphate buffer solution of the same volume, and standing at room temperature for 25min to form a control mixed liquid;
(7) the antiviral effect of the bacterial liquid is detected by adopting a 'half-leaf method': uniformly spreading quartz sand on Sansheng tobacco leaves by taking Sansheng tobacco (NN) cultured for 4 weeks as an experimental material, dipping the mixed solution by using a sterile cotton swab, and slightly rubbing half leaves by taking a main vein as a boundary; the other half of the leaves are rubbed and controlled to form seed mixture;
(8) rubbing for 25min, spraying sterile water to clean the leaves, and naturally air drying at room temperature;
the other steps were the same as in example 1.
In conclusion, the invention discloses a method for separating tobacco mosaic virus resistant bacteria from soil, which separates bacteria with inhibitory effect on tobacco mosaic virus from samples collected from different habitats (shallow sea mud and tobacco rhizosphere soil), enriches the types of beneficial bacteria with antiviral effect, further simply determines the antiviral effect, and lays a foundation for the utilization of the tobacco mosaic virus resistant bacteria.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (10)
1. A method for isolating tobacco mosaic virus-resistant bacteria from soil, comprising the steps of:
(1) taking 1.0g of soil sample, placing into a bottle, adding sterile water, sealing, and culturing for 1-3h with shaking;
(2) standing the solution subjected to shake culture in the step (1) for 20-60min, taking supernatant for gradient dilution, uniformly mixing, and respectively uniformly coating 80-120 mu L of each concentration on a flat plate of a culture medium;
(3) culturing the flat plate in a constant temperature incubator at 25-30 ℃ for 15-20 h;
(4) taking out the cultured plate, selecting a plate with uniformly distributed bacterial colonies from the plate, picking out a single spot, placing the single spot into 2-5mL of culture solution, and performing shake culture for 10-20h to form a bacterial solution;
(5) centrifuging 1mL of bacterial liquid for 8-12min, mixing the supernatant with the isovolumetric tobacco mosaic virus inoculation liquid, and standing at room temperature for 15-30min to form a mixed liquid;
(6) mixing tobacco mosaic virus inoculation liquid with an equal volume of phosphate buffer solution, and standing at room temperature for 15-30min to form a control mixed liquid;
(7) uniformly spreading quartz sand on the Sansheng tobacco leaves by taking the Sansheng tobacco as an experimental material, and slightly rubbing the mixed liquid on half leaves by taking the main veins of the Sansheng tobacco leaves as a boundary; the other half of the leaves are rubbed to contrast the mixed solution;
(8) rubbing the mixed solution and the control mixed solution for 15-30min, spraying sterile water to clean the leaves, and naturally air-drying at room temperature;
(9) carrying out isolation culture in a greenhouse for 2-4d, then counting the number of dead spots on leaves, carrying out one-factor variance analysis on experimental data, carrying out significant difference analysis, and counting the antiviral effect according to the following formula:
2. the method for separating tobacco mosaic virus-resistant bacteria from soil according to claim 1, wherein 7-12mL of sterile water is added in said step (1).
3. The method for separating tobacco mosaic virus-resistant bacteria from soil according to claim 1, wherein said step (1) and/or step (4) is/are shaken at 100-140rpm at 25-30 ℃.
4. The method for separating tobacco mosaic virus-resistant bacteria from soil according to claim 1, wherein the supernatant of step (2) is diluted by gradient 100、10-1、10-2、10-3、10-4And 10-5。
5. The method for separating tobacco mosaic virus-resistant bacteria from soil according to claim 1, wherein said culture medium is LB solid medium.
6. The method for isolating tobacco mosaic virus-resistant bacteria from soil according to claim 1, wherein said culture medium is LB liquid medium.
7. The method for isolating tobacco mosaic virus-resistant bacteria from soil according to claim 1, wherein said soil sample is taken from shallow sea mud or tobacco rhizosphere soil.
8. The method for isolating tobacco mosaic virus-resistant bacteria from soil according to claim 1, wherein said triorganophosphorus is cultured under conditions of a light intensity of 16 hours, a light intensity of 2000lux, a dark duration of 8 hours, a temperature of 25 ± 1 ℃, and a relative humidity of 60%.
9. The method for isolating tobacco mosaic virus-resistant bacteria from soil according to claim 8, wherein said step (7) employs three weeks of incubation of the third-generation smoke.
10. The method for isolating tobacco mosaic virus-resistant bacteria from soil according to claim 1, wherein said tobacco mosaic virus inoculum is prepared by a method comprising the steps of:
(1) taking part of tobacco leaves infected with tobacco mosaic virus, grinding with 10mmol/L, pH 7.4.4 phosphate buffer solution, and homogenizing;
(2) filtering the serous fluid generated in the step (1) by using double-layer gauze, and centrifuging;
(3) taking the supernatant centrifuged in the step (2), treating the supernatant with polyethylene glycol twice, and then centrifuging again;
(4) and (4) after the final centrifugation in the step (3), taking the precipitate, and carrying out heavy suspension by using 10mmol/L, pH 7.4.4 phosphate buffer solution to obtain the tobacco mosaic virus inoculation solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910981804.7A CN110628626A (en) | 2019-10-16 | 2019-10-16 | Method for separating tobacco mosaic virus resistant bacteria from soil |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910981804.7A CN110628626A (en) | 2019-10-16 | 2019-10-16 | Method for separating tobacco mosaic virus resistant bacteria from soil |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110628626A true CN110628626A (en) | 2019-12-31 |
Family
ID=68975131
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910981804.7A Withdrawn CN110628626A (en) | 2019-10-16 | 2019-10-16 | Method for separating tobacco mosaic virus resistant bacteria from soil |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110628626A (en) |
-
2019
- 2019-10-16 CN CN201910981804.7A patent/CN110628626A/en not_active Withdrawn
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108676778B (en) | Bacteriophage for preventing and treating soil-borne bacterial wilt and application thereof | |
| CN108300681B (en) | Streptomyces rochei and application thereof | |
| CN110791448B (en) | Sugarcane endophytic bacillus and application thereof | |
| CN115433697B (en) | Streptomyces siliensis CS13-6 and application thereof | |
| CN114196585B (en) | Burkholderia for preventing and treating tomato bacterial wilt and application thereof | |
| CN115261233B (en) | Biocontrol fungus for stem rot of saffron crocus and application thereof | |
| CN107904196B (en) | Streptomyces yanshi and application thereof | |
| CN111139186B (en) | Trichoderma viride with disease prevention and growth promotion functions and application thereof | |
| CN110628626A (en) | Method for separating tobacco mosaic virus resistant bacteria from soil | |
| CN112410261A (en) | Bacillus siamensis MC2-1 and application thereof | |
| CN110616147A (en) | Process for separating anti-mosaic virus bacteria in soil | |
| CN114561324B (en) | Tomato bacterial wilt antagonistic strain and application thereof in prevention and treatment of tomato bacterial wilt | |
| CN107354112B (en) | Thioanal degrading bacterium and application thereof | |
| CN113940360B (en) | Application of sodium selenite in resisting phytophthora nicotianae | |
| CN114032182B (en) | Fungus with functions of antagonizing pathogenic bacteria of garlic root rot and promoting growth | |
| CN112458021B (en) | Biocontrol pseudomonas fluorescens and culture method and application thereof | |
| EP3622819B1 (en) | Use of a compound for protecting plants by means of stomatal closure and method for protecting plants by means of stomatal closure which comprises applying said compound to the plants | |
| Li et al. | Effects of plant growth regulators on transient expression of foreign gene in Nicotiana benthamiana L. leaves | |
| CN117625475B (en) | Preparation method of microbial metabolite for preventing and treating TMV | |
| CN113201504B (en) | Bacteriophage for preventing and treating plant xanthomonas infection and application thereof | |
| CN115191448B (en) | Microbial plant extract complexing agent for preventing and treating tobacco mosaic virus and application thereof | |
| CN114145300B (en) | Application of salicylic acid and/or p-hydroxybenzoic acid in preparing preparation for preventing and/or treating bacterial soft rot | |
| CN117546873B (en) | Application of extremely oriental pseudomonas in preventing and treating pepper light mottle virus | |
| CN116445376B (en) | Bacillus bailii and application thereof | |
| CN110710537A (en) | Alkaline metalloprotease, preparation containing the same and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20191231 |