CN111202096A - Microbial agent for preventing and treating crop virus diseases and preparation method thereof - Google Patents

Microbial agent for preventing and treating crop virus diseases and preparation method thereof Download PDF

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Publication number
CN111202096A
CN111202096A CN202010035689.7A CN202010035689A CN111202096A CN 111202096 A CN111202096 A CN 111202096A CN 202010035689 A CN202010035689 A CN 202010035689A CN 111202096 A CN111202096 A CN 111202096A
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China
Prior art keywords
parts
powder
microbial agent
virus diseases
preventing
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CN202010035689.7A
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Chinese (zh)
Inventor
胡鉴
王丽
杨军
顾腾腾
顾丽娟
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Henan Nongbeide Agricultural Technology Co ltd
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Henan Nongbeide Agricultural Technology Co ltd
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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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • 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

Abstract

The invention provides a microbial agent for preventing and treating crop virus diseases and a preparation method thereof. The microbial agent comprises the following raw materials in parts by weight: 15-20 parts of bacillus amyloliquefaciens powder, 15-20 parts of bacillus subtilis powder, 15-20 parts of yeast powder, 150 parts of nutrient substances 100-The effective viable count of the microbial inoculum is not less than 2 multiplied by 108cfu/g. The microbial agent contains two effective biocontrol bacteria aiming at the crop virus diseases, and the two effective biocontrol bacteria synergistically generate trans-coumaric acid and apigenin which have a decomposition effect on virus protein shells, so that the crop virus diseases can be effectively prevented and treated.

Description

Microbial agent for preventing and treating crop virus diseases and preparation method thereof
Technical Field
The invention belongs to the technical field of biological products, and particularly relates to a microbial agent for preventing and treating crop virus diseases and a preparation method thereof.
Background
With the development of vegetable production, the diversity of cultivation modes, the richness of vegetable cultivation types and varieties and particularly the diversification of cultivation environments in China, certain secondary diseases rise, the host range of pathogenic bacteria is expanded, a plurality of new diseases to be researched are generated, and a plurality of new and old diseases alternately occur, so that the development of the vegetable industry in China is severely restricted.
The virus diseases of vegetables are serious in occurrence, even cause failure, and become a restrictive obstacle to vegetable production. Viral diseases are diseases caused by parasitic plant viruses. Plant viruses have to live in host cells in a parasitic way, the specificity is strong, and only one or some plants can be infected by one virus. But also a few are widely harmful, such as tobacco mosaic virus and cucumber mosaic virus. Plant viruses are generally active only in the host, and only a few plant viruses can remain active in the diseased plant residues for days, months, or even years, and few plant viruses can survive or proliferate in the insect live. Plant viruses replicate the nucleic acid (RNA or DNA) and protein coat in host cells, constituting novel virions. Plant virus granules or virus nucleic acid have a slow transfer speed among plant cells, and can be rapidly transferred in vascular bundles along with the flowing direction of plant nutrients, so that the plant can be attacked.
At present, chemical agents are mainly used for preventing and treating the diseases, and the chemical agents are used throughout the year, so that the problem of drug resistance is caused, and more seriously, the drug residue greatly threatens the health of human bodies. Therefore, the development of safe, green and pollution-free microbial agents is a trend.
Disclosure of Invention
Problems to be solved by the invention
The invention aims to provide a microbial agent for preventing and treating crop virus diseases and a preparation method thereof.
Means for solving the problems
The invention provides a microbial agent for preventing and treating crop virus diseases. The microbial agent comprises the following raw materials in parts by weight: 15-20 parts of bacillus amyloliquefaciens powder, 15-20 parts of bacillus subtilis powder, 15-20 parts of yeast powder, 150 parts of nutrient substances and 28000 parts of water 25000, wherein the effective viable count in the microbial agent is not less than 2 multiplied by 108cfu/g。
In the above technical scheme, when the microbial agent is applied to crops, the bacillus amyloliquefaciens and the bacillus subtilis exert a synergistic effect, metabolites of the bacillus amyloliquefaciens and the bacillus subtilis react together to generate abundant trans-coumaric acid and apigenin, the trans-coumaric acid and the apigenin have strong decomposition effect on protein shells of viruses, and after the trans-coumaric acid and the apigenin are contacted with the protein shells of the viruses, hyphae gradually surround the surfaces of the viruses, so that the virus proteins are decomposed and lose physiological activity due to the activity of the trans-coumaric acid and the apigenin, and further lose infection, pathogenicity and reproductive capacity. Upon fermentation, the yeast is able to break down the nutrients to produce more small molecule nutrients to supply the metabolism of the bacillus amyloliquefaciens and the bacillus subtilis.
The invention has no special limitation on the types of the nutrient substances, and the nutrient substances are only required to be capable of providing effective viable count for the fermentation and propagation of the bacillus amyloliquefaciens, the bacillus subtilis and the microzyme. Preferably, the nutrient is one or more of fish protein, earthworm protein or bean protein, and more preferably fish protein. The fish protein fertilizer contains rich organic matters, greatly increases the content of the organic matters in the soil after being applied to the soil, can quickly promote the propagation of soil microorganisms, activates the nutrients of the soil to a great extent, increases the fertility of the soil, improves the fertilizer and water retention capacity of the soil, can effectively improve the soil hardening problem by using the fish protein fertilizer, and keeps the soil fertility always new.
Preferably, the microbial agent comprises the following raw materials in parts by weight: 16-18 parts of bacillus amyloliquefaciens powder, 16-18 parts of bacillus subtilis powder, 16-18 parts of yeast powder, 120 parts of nutrient substances and 26000 parts of water. More preferably, the microbial agent comprises the following raw materials in parts by weight: 18 parts of bacillus amyloliquefaciens powder, 18 parts of bacillus subtilis powder, 16 parts of yeast powder, 120 parts of nutrient substances and 25000 parts of water. By further optimizing the proportion, the microbial agent can generate more trans-coumaric acid and apigenin after being applied.
Preferably, in the bacillus amyloliquefaciens powder, the effective viable count of the bacillus amyloliquefaciens is not less than 1 multiplied by 1011cfu/g. In the bacillus subtilis powder, the effective viable count of bacillus subtilis is not less than 1 multiplied by 1011cfu/g. In the yeast powder, the effective viable count of yeast is not less than 5 × 109cfu/g. The three strains can obtain sufficient total effective viable count after fermentation, so that the three strains can play a remarkable synergistic effect to jointly inhibit the growth and the propagation of viruses.
In addition, the invention also provides a preparation method of the microbial agent for preventing and treating the crop virus diseases. The preparation method comprises the following steps: and adding the bacillus amyloliquefaciens powder, the bacillus subtilis powder, the yeast powder, the nutrient substances and water into a fermentation tank, uniformly stirring, and carrying out aerobic fermentation for 45-50 hours at the temperature of 22-27 ℃ and the pH value of 6.5-7.0 to obtain the microbial agent.
Preferably, the aerobic fermentation is carried out for 48 hours at 25 ℃.
In addition, the invention also provides application of the microbial agent for preventing and treating the crop virus diseases in preventing and treating the crop virus diseases.
ADVANTAGEOUS EFFECTS OF INVENTION
(1) The microbial agent contains two effective biocontrol fungi aiming at crop virus diseases, and the two effective biocontrol fungi generate trans-coumaric acid and apigenin which have decomposition effect on virus protein shells under the synergistic effect, so that the crop virus diseases can be effectively prevented and treated;
(2) in the agricultural microbial agent, various microorganisms are easily available, the fermentation method is simple, the period is short, the agricultural microbial agent can be applied to the prevention and the treatment of various crop nematode diseases, and the application range is wide;
(3) the prevention and treatment effect of the microbial agent on the virus diseases of solanaceous vegetables is up to more than 80%.
Detailed Description
The invention will be further described in the following with specific preferred embodiments. In particular, the embodiments of the present invention are not limited thereto, and the following specific examples do not limit the technical aspects of the present invention.
Example 1
18g of active bacteria with the number of 1 multiplied by 1011cfu/g bacillus amyloliquefaciens powder, 18g effective viable count of 1 multiplied by 1011cfu/g bacillus subtilis powder, 16g effective viable count of 5 multiplied by 109Adding cfu/g yeast powder, 120g fish protein and 25kg water into a fermentation tank, stirring uniformly, and performing aerobic fermentation at 22-27 deg.C and pH of 6.5-7.0 for 45-50 hr to obtain the microbial agent. The total effective viable count of the microbial inoculum is detected to be 2.4 multiplied by 108cfu/g。
Example 2
16g of effective viable bacteria with the number of 1 multiplied by 1011cfu/g bacillus amyloliquefaciens powder, 16g effective viable count of 1 multiplied by 1011cfu/g bacillus subtilis powder, 18g effective viable count of 5 multiplied by 109Adding cfu/g yeast powder, 120g earthworm protein and 25kg water into a fermentation tank, stirring uniformly, and performing aerobic fermentation for 45-50 hours at the temperature of 22-27 ℃ and the pH value of 6.5-7.0 to obtain the microbial agent. The total effective viable count of the microbial inoculum is detected to be 2.2 multiplied by 108cfu/g。
Example 3
15g of effective viable bacteria with the number of 1 multiplied by 1011cfu/g bacillus amyloliquefaciens powder, 15g effective viable count of 1 multiplied by 1011cfu/g bacillus subtilis powder, 15g effective viable count of 5 multiplied by 109Adding cfu/g yeast powder, 100g soybean protein and 25kg water into a fermentation tank, stirring uniformly, and performing aerobic fermentation at 22-27 deg.C and pH of 6.5-7.0 for 45-50 hr to obtain the microbial agent. The total effective viable count of the microbial agent is detected as2×108cfu/g。
Comparative example 1
36g of active bacteria with the number of 1 multiplied by 1011cfu/g bacillus subtilis powder, 16g effective viable count of 5 multiplied by 109Adding cfu/g yeast powder, 120g fish protein and 25kg water into a fermentation tank, stirring uniformly, and performing aerobic fermentation at 22-27 deg.C and pH of 6.5-7.0 for 45-50 hr to obtain the microbial agent. The total effective viable count of the microbial inoculum is detected to be 2 multiplied by 108cfu/g。
Comparative example 2
36g of active bacteria with the number of 1 multiplied by 1011cfu/g bacillus amyloliquefaciens powder, 16g effective viable count of 5 multiplied by 109Adding cfu/g yeast powder, 120g fish protein and 25kg water into a fermentation tank, stirring uniformly, and performing aerobic fermentation at 22-27 deg.C and pH of 6.5-7.0 for 45-50 hr to obtain the microbial agent. The total effective viable count of the microbial inoculum is detected to be 2 multiplied by 108cfu/g。
Comparative example 3
The effective viable count of 12g is 1 multiplied by 1011cfu/g bacillus amyloliquefaciens powder, 13g effective viable count of 1 multiplied by 1011cfu/g bacillus amyloliquefaciens powder, 13g effective viable count of 5 multiplied by 109Adding cfu/g yeast powder, 120g fish protein and 25kg water into a fermentation tank, stirring uniformly, and performing aerobic fermentation at 25 deg.C and pH of 6.5 for 45-50 hr to obtain the microbial preparation. The total effective viable count of the microbial inoculum is 1.4 multiplied by 108cfu/g。
Field efficacy test
Tomato fields with consistent growth vigor and consistent occurrence of yellow leaf curl disease are selected, the agricultural microbial agents obtained in the examples 1-3 and the comparative examples 1-3 are sprayed at the amount of 300 mL/mu, and the statistical conditions of the control effect are shown in the table 1 after the agricultural microbial agents are applied for half a month and one month.
TABLE 1 field efficacy test results of examples 1-3 and comparative examples 1-3
Figure BDA0002365905210000051
As can be seen from Table 1, the microbial agents in the examples 1 to 3 have obvious control effect on tomato yellow leaf curl disease, and the control effect is over 96 percent after the microbial agents are sprayed for 1 month. The microbial agent of the comparative example 1 is not added with bacillus amyloliquefaciens, and the microbial agent of the comparative example 2 is not added with bacillus subtilis, so that the synergistic effect of the bacillus amyloliquefaciens and the bacillus subtilis can not be exerted, and the control effect is not achieved. The microbial agent of the comparative example 3 is added with the bacillus amyloliquefaciens and the bacillus subtilis in lower parts by weight, so that the total effective viable count after fermentation is greatly reduced, and the prevention effect of the obtained microbial agent is also reduced.

Claims (10)

1. A microbial agent for preventing and treating crop virus diseases is characterized by comprising the following raw materials in parts by weight: 15-20 parts of bacillus amyloliquefaciens powder, 15-20 parts of bacillus subtilis powder, 15-20 parts of yeast powder, 150 parts of nutrient substances and 28000 parts of water 25000, wherein the effective viable count in the microbial agent is not less than 2 multiplied by 108cfu/g。
2. The microbial agent for preventing and treating crop virus diseases according to claim 1, which is characterized by comprising the following raw materials in parts by weight: 16-18 parts of bacillus amyloliquefaciens powder, 16-18 parts of bacillus subtilis powder, 16-18 parts of yeast powder, 120 parts of nutrient substances and 26000 parts of water.
3. The microbial agent for preventing and treating crop virus diseases according to claim 2, which is characterized by comprising the following raw materials in parts by weight: 18 parts of bacillus amyloliquefaciens powder, 18 parts of bacillus subtilis powder, 16 parts of yeast powder, 120 parts of nutrient substances and 25000 parts of water.
4. The microbial agent for preventing and treating crop virus diseases according to claim 1 or 2, wherein the effective viable count of bacillus amyloliquefaciens in the bacillus amyloliquefaciens powder is not less than 1 x 1011cfu/g。
5. The microbial agent for controlling crop viral diseases according to claim 1 or 2, wherein the effective viable count of Bacillus subtilis in the Bacillus subtilis powder is not less than 1 x 1011cfu/g。
6. The microbial agent for controlling crop virus diseases according to claim 1 or 2, wherein the effective viable count of yeast in the yeast powder is not less than 5 x 109cfu/g。
7. The microbial agent for preventing and treating crop virus diseases according to claim 1 or 2, wherein the nutrient is one or more of fish protein, earthworm protein or bean protein.
8. The method for preparing microbial agents for controlling crop virus diseases according to any one of claims 1 to 7, comprising the steps of: and adding the bacillus amyloliquefaciens powder, the bacillus subtilis powder, the yeast powder, the nutrient substances and water into a fermentation tank, uniformly stirring, and carrying out aerobic fermentation for 45-50 hours at the temperature of 22-27 ℃ and the pH value of 6.5-7.0 to obtain the microbial agent.
9. The method for preparing microbial inoculum for preventing and treating crop virus diseases as claimed in claim 8, characterized in that aerobic fermentation is carried out for 48 hours at 25 ℃.
10. The use of a microbial agent for controlling crop virus diseases according to any one of claims 1 to 7 for controlling crop virus diseases.
CN202010035689.7A 2020-01-14 2020-01-14 Microbial agent for preventing and treating crop virus diseases and preparation method thereof Pending CN111202096A (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000051435A1 (en) * 1999-03-01 2000-09-08 Auburn University Biological compositions and methods for enhancing plant growth and health and producing disease-suppressive plants
CN106793782A (en) * 2014-08-06 2017-05-31 恩威拉有限责任公司 For the bacterial spore composition of industrial use
WO2019067380A2 (en) * 2017-09-28 2019-04-04 Locus Agriculture Ip Company, Llc Treatment of mosaic viruses and bacterial infections of plants

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000051435A1 (en) * 1999-03-01 2000-09-08 Auburn University Biological compositions and methods for enhancing plant growth and health and producing disease-suppressive plants
CN106793782A (en) * 2014-08-06 2017-05-31 恩威拉有限责任公司 For the bacterial spore composition of industrial use
WO2019067380A2 (en) * 2017-09-28 2019-04-04 Locus Agriculture Ip Company, Llc Treatment of mosaic viruses and bacterial infections of plants

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
国家医药管理局中草药情报中心站 编: "《植物药有效成分手册》", 31 March 1986, 人民卫生出版社 *
林辉 主编: "《有机化学》", 31 January 2017, 中国中医药出版社 *

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Application publication date: 20200529