CN113151119A - Paenibacillus terrestris for preventing and treating gray mold of paris polyphylla and application thereof - Google Patents
Paenibacillus terrestris for preventing and treating gray mold of paris polyphylla and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/25—Paenibacillus
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- 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
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Abstract
The invention belongs to the technical field of plant disease and insect pest control, and particularly relates to paenibacillus terricola for controlling paris polyphylla gray mold and application thereof. The Paenibacillus terrestris strain provided by the invention is derived from the interior of the leaves of the Paris polyphylla, and has good compatibility with a self-circulation system of the Paris polyphylla; the microbial inoculum is simple to prepare, low in cost, non-toxic, harmless and free of environmental pollution, is beneficial to safely and effectively controlling the gray mold of the paris polyphylla, improves the yield and the quality of the paris polyphylla, is beneficial to green cultivation of the paris polyphylla, protects a farmland ecosystem and promotes the sustainable development of the paris polyphylla industry. The microbial inoculum is beneficial to improving the planting soil of the paris polyphylla, increasing the microbial diversity in the soil and promoting the output and the effective component content of the paris polyphylla.
Description
Technical Field
The invention belongs to the technical field of plant disease and insect pest control, and particularly relates to paenibacillus terricola for controlling paris polyphylla gray mold and application thereof.
Background
Paris polyphylla (Paris polyphylla) is a liliaceae medicinal plant, is widely distributed in Sichuan, Yunnan, Guizhou, Hunan, Hubei and other places in China, has the effects of inhibiting bacteria, clearing heat and removing toxicity, relieving swelling and pain and the like, and can be clinically used for calming and easing pain, treating venomous snake bite, treating convulsion and tetany, resisting cancer, protecting a cardiovascular system and the like. In addition, rhizoma paridis is the main raw material of Chinese patent medicines such as Yunnan white drug powder, GONGXUENING, Qudesheng snake drug, etc., and the main active ingredients include pariphyllin, dioscin, steroid and alkaloid, etc. Due to slow growth of the paris polyphylla, and the fact that people abused and disorderly dig for a long time, wild paris polyphylla resources are exhausted and cannot meet market demands, market price of the paris polyphylla rises year by year, and the benefit per mu is nearly 20 ten thousand yuan in recent years. With the attention of governments at all levels to the biological medicine and health industry, the traditional Chinese medicine paris polyphylla becomes the main industry of the genuine production area because of various efficacies. In recent years, the artificial planting area of the paris polyphylla is enlarged year by year, the problem of diseases is obvious, and the gray mold of the paris polyphylla (Botrytis cinerea) in Hubei, Hunan, Sichuan and the like shows an outbreak trend, wherein the average incidence rate of diseases of various bases in Hubei province reaches about 55 percent, the serious disease can reach 100 percent, the humidity of the paris polyphylla in each year is 3-5 months, the field humidity of the paris polyphylla is high, and the temperature is proper, so that the period of the most serious gray mold of the paris polyphylla is provided.
The prevention and control of the gray mold are mainly chemical prevention and control at present, and carbendazim, fenhexamid, tolylfluanid, fludioxonil and the like are common agents for preventing and controlling the gray mold. Because the registered traditional Chinese medicinal materials are few in medicament and the phenomenon of no medicament is serious, the frequent use of the pesticide is caused. The abuse of pesticide agents and the long-term use of single agents in actual production cause the problems of pesticide residue exceeding standards, environmental pollution and pathogenic bacteria drug resistance to be prominent, and seriously threaten the quality of the paris polyphylla.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides Paenibacillus terrae for preventing and treating paris polyphylla gray mold and application thereof, and aims to solve part of problems in the prior art or at least alleviate part of problems in the prior art.
The invention is realized in such a way that a Paenibacillus terrae for preventing and treating gray mold of paris polyphylla is named as Paenibacillus terrae PY8(Paenibacillus terrae PY8), is preserved in China Center for Type Culture Collection (CCTCC) in 9-month and 4-month 2020, and has the preservation number: CCTCC NO: m2020462. Address: wuhan university in Wuhan, China.
The invention also provides application of the paenibacillus terricola for preventing and treating the gray mold of the paris polyphylla in preparation of a gray mold preventing and treating microbial inoculum with streptomycin resistance.
The invention also provides a streptomycin resistant botrytis cinerea control microbial inoculum which comprises the paenibacillus terrae.
The invention also provides a preparation method of the streptomycin resistant botrytis cinerea control microbial inoculum, which comprises the steps of inoculating Paenibacillus terrae in an NB liquid culture medium, and carrying out aerobic fermentation at 28 ℃ to obtain bacteriostatic fermentation liquor.
Further, the method comprises the steps of centrifuging the bacteriostatic fermentation liquor, collecting supernatant, filtering with a filter membrane, and mixing the filtrate with NA.
Further, the preparation method comprises the steps of coating Paenibacillus terrae on NA culture medium for activation; inoculating the activated Paenibacillus terrae into an NB liquid culture medium for aerobic fermentation to prepare a fermentation seed solution; and then inoculating the fermentation seed liquid into an NB liquid culture medium for aerobic fermentation to obtain the bacteriostatic fermentation liquid.
Furthermore, the concentration of viable bacteria in the bacteriostatic fermentation liquor reaches 1 × 108CFU/ml above.
Furthermore, the concentration of viable bacteria in the bacteriostatic fermentation liquor reaches 1 × 1010CFU/ml above.
The invention also provides application of the paenibacillus terrae in preventing and treating the gray mold of the paris polyphylla.
The invention also provides application of the Paenibacillus terrae in regulating and controlling the yield of the paris polyphylla and/or the bacterial community structure of the paris polyphylla leaves.
Further, the rhizoma paridis yield includes at least one of fresh weight of single plant root, rhizoma paridis saponin I content and rhizoma paridis saponin II content.
The biological control method for preventing and treating the gray mold of the paris polyphylla has good popularization value and ecological benefit, and no report is found in the research of preventing and treating the gray mold of the paris polyphylla by utilizing the Paenibacillus terrae (Paenibacillus terrae) at home and abroad at present. The strain provided by the invention can be used for preventing and treating the gray mold of the paris polyphylla, and the target strain has an obvious inhibiting effect on gray mold hypha of the paris polyphylla, and can cause the gray mold hypha of the paris polyphylla to expand and deform, be short and cause protoplasm leakage. When the concentration of the target strain reaches 1X 1010The control effect reaches 89.61 percent when CFU/mL is used, and the control effect is equivalent to that of a chemical agent.
In summary, the advantages and positive effects of the invention are:
1. the target strain is from the interior of the leaves of the paris polyphylla, and has good compatibility with the self circulation system of the paris polyphylla.
2. The microbial inoculum is simple to prepare, low in cost, non-toxic, harmless and free of environmental pollution, and is beneficial to safely and effectively controlling the gray mold of the paris polyphylla.
3. The microbial inoculum is beneficial to improving the planting soil of the paris polyphylla and increasing the diversity of microorganisms in the soil.
4. The microbial inoculum is a biological preparation, has no defects of chemical pesticides, can not cause pathogenic bacteria to generate drug resistance, a grower can not use or reduce the use frequency of other chemical pesticides, and the biocontrol strain preparation has the functions of increasing yield and efficiency and can increase income of farmers.
5. The microbial inoculum can promote the output and the content of effective components of the rhizoma paridis to a certain degree.
The plate antagonism experiment and the antagonism experiment of fermentation supernatant with different concentrations show that: the target microbial inoculum has an obvious inhibiting effect on the botrytis cinerea hyphae, and can cause the botrytis cinerea hyphae to expand and deform, be exhausted and cause protoplasm leakage. The field control experiment proves that when the concentration of the thallus reaches 1 multiplied by 10, the target microbial inoculum is on the overground part of the sprayed plant10The control effect of CFU/mL reaches 8961%, and the tuber yield of the paris polyphylla can be increased by about 5.77% compared with the control, and the paris saponin I and the paris saponin II can be increased by 0.83% and 1.18% respectively compared with the control.
Drawings
FIG. 1 shows the effect of target bacteria on the growth of gray mold hyphae of Paris polyphylla; in the figure: the left is a confrontation graph of target bacteria and paris polyphylla botrytis; the right part is the comparison of Botrytis cinerea.
FIG. 2 is the effect of target bacteria on Gray mold bacteria hyphae during co-cultivation; in the figure: a is the hypha form of botrytis cinerea in the opposite area of co-culture; b is a comparison;
FIG. 3 is a graph showing the effect of different concentrations of target bacteria fermentation filtrates on the growth of Gray mold bacteria;
FIG. 4 shows the control effect of target bacteria for field application;
FIG. 5 is the colonization of the leaves by the target bacteria;
FIG. 6 is a table of microbial diversity indices for different treatment populations;
FIG. 7 is a statistical table of populations of different treated microorganisms;
FIG. 8 is a heat map of the genus Paris polyphylla leaf bacteria classification.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples, and the equipment and reagents used in the examples and test examples are commercially available without specific reference. The specific embodiments described herein are merely illustrative of the invention and are not intended to be limiting.
Various modifications to the precise description of the invention will be readily apparent to those skilled in the art from the information contained herein without departing from the spirit and scope of the appended claims. It is to be understood that the scope of the invention is not limited to the procedures, properties, or components defined, as these embodiments, as well as others described, are intended to be merely illustrative of particular aspects of the invention. Indeed, various modifications of the embodiments of the invention which are obvious to those skilled in the art or related fields are intended to be covered by the scope of the appended claims.
For a better understanding of the invention, and not as a limitation on the scope thereof, all numbers expressing quantities, percentages, and other numerical values used in this application are to be understood as being modified in all instances by the term "about". Accordingly, unless expressly indicated otherwise, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. In the present invention, "about" means within 10%, preferably within 5% of a given value or range.
In the following examples of the present invention, the temperature is not particularly limited, and all of the conditions are normal temperature conditions. The normal temperature refers to the natural room temperature condition in four seasons, no additional cooling or heating treatment is carried out, and the normal temperature is generally controlled to be 10-30 ℃, preferably 15-25 ℃.
The invention discloses paenibacillus terricola for preventing and treating gray mold of paris polyphylla and application thereof, and particularly relates to the following embodiment.
Example 1 isolation, screening and identification of target strains
Separating target strain from leaves of rhizoma paridis of Enshi city of Hubei province, sterilizing the leaves with 75% alcohol for 1min, washing with sterile water for 3 times, air drying under sterile condition, grinding with sterile water, diluting the mixed solution, spreading on NA plate, culturing at 28 deg.C for 72 hr, and selecting single colony for purification and culture. And 3cm away from botrytis cinerea, placing endophytes of the leaves of the paris polyphylla obtained by separation according to a cross method, after 3 days, measuring the distance from the outer edge of a bacterial colony to botrytis cinerea hyphae by contrasting and growing in a dish, repeating for 3 times, and selecting the bacteria with the largest distance for later research.
Through detection, the target bacterial strain is gram-positive bacteria, the thallus is rod-shaped, and spores and periphytic flagella can be formed. The bacterial strain is cultured in NA for 7d to form a round bacterial colony with the diameter of about 2mm, the middle of a single bacterial colony is wrinkled, the surface is wet and transparent, the bacterial lawn is thick and is easy to pick up. The later colony edge of the strain is irregular, and IAA can be produced.
And (2) coating 20 mu L of fermentation liquor of a target strain on an NA (N-acetyl-D) plate containing 10 mu g/mL streptomycin, culturing for 2d at 28 ℃, picking a single colony, inoculating the single colony into 100mL of NB medium containing 10 mu g/mL streptomycin, culturing for 2d at 28 ℃, 200r/min, picking the single colony, transferring the single colony to the next concentration culture, inducing in NB medium containing 20, 40, 80, 160, 200 and 300 mu g/mL of streptomycin once until a mutant strain resisting 300 mu g/mL of streptomycin and stably growing is screened, continuously passaging the streptomycin-resisting strain on the NB medium without antibiotics for 5 times, and then inoculating the mutant strain into the medium containing the streptomycin for detection to ensure the genetic stability of drug resistance. Meanwhile, the bacterial colony morphology, physiological and biochemical characteristics and antagonistic action on pathogenic bacteria of the strain are kept stable.
Through physiological, biochemical and sequencing analysis and identification, the strain is Paenibacillus terreae PY8, is preserved in China center for type culture Collection in 9-month and 4-month 2020, and has the preservation number: CCTCC NO: m2020462. Address: wuhan university in Wuhan, China.
Example 2 preparation of the objective microbial Agents
S1, taking one loop of the target strain by using an inoculating loop, coating the loop on an NA culture medium, and culturing for 72 hours at 28 ℃. The formula of the NA solid culture medium is as follows: taking 5.0g of beef extract, 10.0g of peptone, 5g of NaCl, 15g of agar and 1000mL of purified water, carrying out high-pressure steam sterilization at 121 ℃ for 30min at pH 7.2-7.4.
S2, transferring the target single colony activated in the step S1 into a 250ml conical flask of 100ml NB liquid medium, and culturing at 200rmp and 28 ℃ for 72 hours to obtain a fermentation seed solution. The NB liquid medium formula is as follows: taking 5.0g of beef extract, 10.0g of peptone, 5g of NaCl and 1000mL of purified water, and sterilizing for 30min at 121 ℃ by high-pressure steam, wherein the pH is 7.2-7.4.
S3, inoculating the fermentation seed liquid in the step S2 into a conical flask containing 100ml of NB according to the ratio of 1:150(V: V), placing the conical flask in a vibrator, fermenting at 28 ℃ and 200rmp for 72h to ensure that the concentration of viable bacteria reaches 1 x 1010More than CFU/ml, and then storing the produced microbial inoculum in a refrigerator at 4 ℃.
Plate confrontation antagonism test
According to a plate confronting culture test method in the formula of 'plant disease research method', the inhibition effect of target bacteria on botrytis cinerea is evaluated, and meanwhile, the bacteria hyphae of a sterilized distilled water control treatment group are counted when the plates are full of bacteria hyphae, and the inhibition rate (%) is (the diameter of the control group pathogenic bacteria colony-the diameter of the treatment group pathogenic bacteria colony)/the diameter of the control group pathogenic bacteria colony is multiplied by 100%. The results demonstrated that the target bacteria had an inhibition rate of 72.22% on the growth of botrytis cinerea hyphae (as shown in fig. 1).
Microscopic observation of botrytis cinerea hyphae in confrontation area
And observing the hypha form of the botrytis cinerea by using a microscope in the confronting area of the target fungi and the botrytis cinerea hypha. The results showed slow growth of hyphae, thin hyphae, malformation, and leakage of protoplasts of Gray mold of Paris polyphylla in the confronting areas (as shown in FIG. 2).
EXAMPLE 3 Effect of fermentation supernatant of target bacteria on Gray mold of Paris polyphylla
And (3) injecting 500 mu L of activated target bacterial liquid into 100mL of NB, and culturing at 28 ℃ and 180r/min for 72h to obtain a fermentation stock solution. Centrifuging the fermentation stock solution at 12000r/min for 10min, collecting supernatant, and filtering with 0.22 μm microporous membrane to obtain fermentation filtrate. Mixing the filtrate with PDA at about 45 deg.C to give final concentration of 5% and 10%, cooling, inoculating Botrytis cinerea mycelium block (diameter 0.5cm) at the center of the plate, setting blank control, repeating the treatment three times, and culturing at 21 deg.C in incubator. After the control was plated over (4 days), the diameter of the colony was measured and the inhibition rate was calculated. The inhibition ratio (%) - (control group pathogen colony diameter-treatment group pathogen colony diameter)/control group pathogen colony diameter × 100%.
The results prove that the target bacteria fermentation filtrate can obviously inhibit the growth of botrytis cinerea at both concentrations of 5% and 10%, the inhibition rates of the target bacteria fermentation filtrate and the target bacteria fermentation filtrate are 44.4% and 74.44%, and the inhibition rates of the target bacteria fermentation filtrate and the target bacteria fermentation filtrate are obviously different (as shown in figure 3).
Example 4 field control of Gray mold of Paris polyphylla by target bacteria
The test point is arranged in a Paris polyphylla resource garden in a new pond, countryside and China medicinal plant garden in Enshi City of Hubei province. At an altitude of 1600m, gray mold occurs in the last year, the separated pathogenic bacteria are mainly botrytis cinerea, all operations are consistent in the planting process, other chemical agents are not used, and the paris polyphylla is a 4-year-old plant and can normally bloom and bear fruits. The test time is 3 months and 10 days in 2018, the weather is sunny, the rate of emergence of the paris polyphylla is 95 percent, and the paris polyphylla just emerges for 2 weeks.
Diluting the fermentation stock solution of the target strain to 1 × 1010CFU/mL、1×109CFU/mL and 1X 108CFU/mL is reserved, 1000 times of liquid of 50% iprodione wettable powder serving as a chemical agent, 800 times of liquid of 1000 hundred million spores/gram bacillus subtilis wettable powder serving as a biological agent serve as positive control, and clear water serves as negative control. Spraying the whole plant and the soil surface according to the recommended concentration of each medicament at 9 noon of 2018, 3-month 10, 3-month 16, 3-month 22 and 3-month 28, and continuously applying for 4 times, wherein no rain exists during the application period and 12 hours after the application. And investigating the influence of the treatment on the paris polyphylla in 2018, 4 and 17 days, sampling five points in each cell, taking 5 plants in each point, counting the fresh weight of rhizomes and the contents of paris polyphylla saponin I and paris polyphylla saponin II, and calculating the average weight and content of the rhizomes. And calculating the control effect of each treatment according to the grading standard. Grading standard: grade 0, no disease; grade 1, the affected area accounts for less than 25% of the total area of the fruit; grade 3, the affected area accounts for 25-50% of the total area of the fruit; grade 5, the affected area accounts for 50-75% of the total area of the fruit; grade 7, the affected area accounts for more than 75% of the total area of the fruit (Zhang J et al, Mycoscience,2010,51(6): 421.). The disease index is 100 × Σ (number of diseased plants at each stage × representative value at each stage)/(total number of plants × highest representative value at each stage); control effect (%) < 100 × (control disease index-treatment disease index)/control disease index. The experimental result proves that at 20 days after the drug administration, the concentration of the endophytic target strain in the paris polyphylla is 1 multiplied by 10 in three concentrations10The control effect of CFU/mL treatment reaches 89.61 percent, 1 is multiplied by 109The treatment control effect of CFU/mL is 81.37%, 1 × 108The control effect of CFU/mL treatment is 73.73% (shown in figure 4), wherein the control effect of the commercial microbial agent Bacillus subtilis is 59.22%, and the control effect of the chemical agent iprodione in all treatments is the highest and reaches 90.98%. 1X 10 of the target bacteria in the influence of each treatment on the rhizome weight of Paris polyphylla10The fresh weight of CFU/mL treatment was 18.19g at the highest, followed by 1X 109The fresh weight of the CFU/mL treatment is 18.11g, and the yield is increased by 5.77% and 5.36% respectively compared with the control. At each location1X 10 of target bacteria in the detection of treatment content10CFU/mL treatment, the content of rhizoma paridis saponin I and rhizoma paridis saponin II is 0.1803% and 0.4322%, respectively, which are higher than those of control treatment, and the content of rhizoma paridis saponin I and rhizoma paridis saponin II in target bacteria is 1 × 109In the CFU/mL treatment, the contents of the paris polyphylla saponin I and the paris polyphylla saponin II are respectively 0.1792% and 0.4311%, which are also higher than a control, and the difference does not reach a very significant level. The yield can be improved to a certain extent after the target microbial inoculum is applied, and the contents of the rhizoma paridis saponin I and the rhizoma paridis saponin II are improved. Specific data are shown in table 1.
TABLE 1 field control effect of different treatments on gray mold of Paris polyphylla
In the table, different lower case letters in the same column of data indicate that the difference is significant and P is less than 0.05
Example 5 colonization of the leaves by the target Strain
3.10 and 3.16 of 2018, 1X 1010Uniformly spraying the fermentation liquor of the target strain of the CFU/mL for 2 times, respectively taking 1, 3, 5, 7, 10, 15, 20, 30, 40, 50, 60 and 80d after inoculation, washing each sample in sterile water for 6 times, taking 0.5g of the sample, disinfecting the sample by using 75% alcohol for 30S, then disinfecting the sample by using 1% sodium hypochlorite for 3min, then washing the sample by using sterile water for 5 times, airing the sample in a sterile operating platform, adding 1mL of water into a sterile mortar for grinding, sucking 100 mu L of supernatant after standing for 10min, sequentially diluting the supernatant for 10, 100 and 1000 times, then sequentially coating the diluted supernatant on an NA plate of streptomycin 300 mu g/mL, repeating the steps for 3 times, culturing the product at 28 ℃ for 48h, counting the number of bacterial colonies, and calculating the number of bacteria (CFU/g) in each gram of tissue according to the number of bacterial colonies appearing on each dish. Meanwhile, randomly picking out grown colonies for morphological, physiological and biochemical analysis and 16SrDNA identification, and confirming the colonies to be Paenibacillus terreus (Paenibacillus terrae).
Through statistics, the target strain can still be detected on the leaves of the paris polyphylla till 60d, a colonization peak appears 7 days after spraying, and the bacterial quantity reaches 4.2 multiplied by 105CFU/g, followed by a downward trend, about 1X 1 of leaf bacterial load at day 6003CFU/g, it can be seen that the target strain can be well colonized on the leaves of the Paris polyphylla. (see FIG. 5 for a detailed profile).
Example 6 Effect of target strains on the bacterial colony Structure of Paris polyphylla leaves
Fermenting the target bacteria at 1 × 1010After CFU/mL spraying Hua re-recording the leaves, sampling is carried out after 10 days, and the bacterial flora structure of the leaves is detected by an Illumina MiSeq sequencing platform in 2017 by utilizing. The sequencing results show that after the target bacteria are used, the Simpson index, the Chao1 index, the ACE index and the Shannon index of the paris polyphylla leaves are all higher than those of the blank control, wherein the Simpson index and the Shannon index are 3 times higher than those of the control (shown in figure 6). Meanwhile, after the target bacteria are treated, the number of the seed level is higher than that of a blank control, the control of other indexes is relatively higher (shown in figure 7), and heat map analysis is carried out on the genus level aiming at different treatments, so that the pseudomonas treated by the target bacteria is 70 times that of the control, the moraxelleaceae is 102 times that of the control, and the Flavobacteriaceae and the Enterobacteriaceae exist in the treatment of the target bacteria as far as possible, but do not exist in a clear water control. In the first 50 genus classification heatmaps shown, 30 species, 60% of the target bacteria were dominant, and it was seen that the target bacteria had the property of stimulating the growth of some bacteria (FIG. 8).
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (10)
1. The paenibacillus terricola for preventing and treating the gray mold of the paris polyphylla has the preservation number of CCTCC NO: m2020462, titled Paenibacillus terrae (Paenibacillus terrae PY 8).
2. The application of the paenibacillus terricola for preventing and treating the gray mold of the paris polyphylla as claimed in claim 1 in preparing a gray mold preventing and treating microbial inoculum with streptomycin resistance.
3. A botrytis cinerea control microbial inoculum with streptomycin resistance is characterized in that: comprising Paenibacillus terrae as claimed in claim 1.
4. The method for preparing a streptomycin-resistant gray mold control bacterial agent as claimed in claim 3, wherein the method comprises the following steps: comprises inoculating Paenibacillus terrestris in NB liquid culture medium, and aerobically fermenting at 28 deg.C to obtain antibacterial fermentation liquid.
5. The method for preparing a botrytis cinerea bacterial agent with streptomycin resistance as claimed in claim 4, wherein the method further comprises the steps of centrifuging bacteriostatic fermentation liquor, collecting supernatant, filtering with a filter membrane, and mixing the filtrate with NA.
6. The method for preparing a streptomycin-resistant botrytis cinerea control bacterium agent as claimed in claim 4, wherein the preparation method comprises activating Paenibacillus terrae spread on a NA medium; inoculating the activated Paenibacillus terrae into an NB liquid culture medium for aerobic fermentation to prepare a fermentation seed solution; and then inoculating the fermentation seed liquid into an NB liquid culture medium for aerobic fermentation to obtain the bacteriostatic fermentation liquid.
7. The method for preparing a streptomycin resistant botrytis cinerea prevention and treatment microbial inoculum according to claim 4, wherein the concentration of viable bacteria in the bacteriostatic fermentation liquid reaches 1 x 108CFU/ml above.
8. The method for preparing a streptomycin resistant botrytis cinerea prevention and treatment microbial inoculum according to claim 7, wherein the concentration of viable bacteria in the bacteriostatic fermentation liquid reaches 1 x 1010CFU/ml above.
9. Use of Paenibacillus terrae as claimed in claim 1 for the control of Paris polyphylla gray mold.
10. Use of Paenibacillus terrae as claimed in claim 1 for regulating the production and/or structure of the bacterial community of Paris polyphylla leaves.
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| CN109112087A (en) * | 2018-08-21 | 2019-01-01 | 甘肃省农业科学院植物保护研究所 | One plant of soil series bacillus YC16-08 and its application |
| CN110628686A (en) * | 2019-10-28 | 2019-12-31 | 云南省农业科学院生物技术与种质资源研究所 | Bacillus polymyxa strain and application thereof |
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