CN109169710B - Biocontrol microbial inoculum for preventing and treating root-knot nematodes and application thereof - Google Patents

Biocontrol microbial inoculum for preventing and treating root-knot nematodes and application thereof Download PDF

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CN109169710B
CN109169710B CN201811168393.1A CN201811168393A CN109169710B CN 109169710 B CN109169710 B CN 109169710B CN 201811168393 A CN201811168393 A CN 201811168393A CN 109169710 B CN109169710 B CN 109169710B
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bacillus licheniformis
pseudomonas fluorescens
phanerochaete chrysosporium
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董海龙
路平
杜宾
王琳
徐玉梅
张作刚
崔克勇
王建明
王慧
杨斌
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Shanxi Agricultural University
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Abstract

The invention belongs to the technical field of microbial pesticides, and provides a biocontrol microbial inoculum for preventing and treating root-knot nematodes and application thereof in order to solve the problems of single disease prevention mechanism and low prevention effect of plants according to a single strain of nematodes at present. The method comprises the following steps of mixing a bacillus licheniformis-pseudomonas fluorescens mixed fermentation liquid, a streptomyces fradiae fermentation liquid and a phanerochaete chrysosporium-rhus verruculosus mixed fermentation liquid according to the weight ratio of 2-5: 1-3: 1-7, and then uniformly mixing the mixed solution and the quinoa peel according to the mass ratio of 1kg to 6-8 kg. The five strains are not antagonistic to each other and are synergistic to each other for disinsection, and the preparation process is relatively simple and the application prospect is wide. The invention can prevent and control vegetable root knot nematode disease, promote plant growth and development and improve plant disease resistance.

Description

Biocontrol microbial inoculum for preventing and treating root-knot nematodes and application thereof
Technical Field
The invention belongs to the technical field of microbial pesticides, and particularly relates to a biocontrol microbial inoculum for preventing and treating root-knot nematodes and application thereof.
Background
Root knot nematode (A), (B) and (C)Meloidogyne spp.) The disease is one of important root diseases which seriously affect the yield of vegetable facility cultivation in the global range, and has the characteristics of wide distribution, multiple hosts, strong infectivity, hidden occurrence of harm and the like. With the rapid development of the facility vegetable cultivation, the incidence rate of root-knot nematodes rises year by year, and the yield of serious areas can be reduced by 30-40%, even the root-knot nematodes are not harvested. At present, the most important prevention and treatment means in agriculture is still pesticide and soil disinfection, which causes a great amount of pesticide residues and environmental pollution. Therefore, the quality and yield of vegetables are improved, and the search for safe, efficient and environment-friendly measures for preventing and controlling the root knot nematode becomes an urgent important demand of the vegetable industry. At present, indoor researches show that pseudomonas fluorescens metabolites, conidia and metabolites of the myrothecium verrucaria and conidia of the phanerochaete chrysosporium have strong killing and parasitic effects on root-knot nematodes J2 and eggs, and are bio-control strains for effectively controlling the root-knot nematodes.
In the application process of biocontrol bacteria, a single microorganism is used for preventing and controlling plant root-knot nematodes, but a single strain has a single disease prevention mechanism and poor definite value capability, is easily influenced by various factors of soil environment, and causes a low prevention and control effect. Through mutual affinity and synergistic effect among the biocontrol bacteria, the advantages of multiple strains are complemented, the growth vigor or yield of the plant can be improved, the population quantity of soil root-knot nematodes is reduced, the quantity of soil free nematodes and beneficial microorganisms is increased, the root of the plant is prevented from being infected by the root-knot nematodes, and the biocontrol effect and stability are improved.
Disclosure of Invention
The invention provides a biocontrol microbial inoculum for preventing and treating root-knot nematodes and application thereof, aiming at solving the problems of single disease prevention mechanism and low prevention and treatment effect of plants according to a single strain of nematodes at present.
The invention is realized by the following technical scheme: a biocontrol microbial inoculum for preventing and treating root-knot nematode is prepared from Bacillus licheniformis (Bacillus licheniformis)Bacillus licheniformis) Pseudomonas fluorescens (Pseudomonas fluorescens) Mixed fermentation broth, Streptomyces fradiae: (Streptomyces fradiae) Fermentation broth, Phanerochaete chrysosporium: (Phanerochaete chrysosporium Burdsall) Myrothecium verrucaria (A. japonicas (A.)), (B. verrucosa (B.))Myrothecium verrucaria) Mixing the fermentation liquor according to the ratio of 2-5: 1-3: 1-7, and then uniformly mixing the mixed solution and the quinoa peel according to the mass ratio of 1kg to 6-8 kg;
wherein: bacillus licheniformis (Bacillus licheniformis) Pseudomonas fluorescens (Pseudomonas fluorescens) The mixed fermentation liquor comprises: the bacillus licheniformis seed solution and the pseudomonas fluorescens seed solution are mixed according to the weight ratio of 1-2: 1, stirring for 3-5 minutes, inoculating into a fermentation culture medium according to the inoculation amount of 3-8% of the volume ratio, and culturing at 30 ℃ for 18-22 hours to obtain a bacillus licheniformis-pseudomonas fluorescens mixed fermentation liquor;
streptomyces fradiae (Streptomyces fradiae) The fermentation liquor is: inoculating the Streptomyces fradiae seed liquid into a fermentation culture medium according to the inoculation amount of 3-8% in volume ratio, and culturing for 7d at 29 ℃ to obtain a Streptomyces fradiae fermentation liquid;
phanerochaete chrysosporium (A)Phanerochaete chrysosporium Burdsall) Myrothecium verrucaria (A. japonicas (A.)), (B. verrucosa (B.))Myrothecium verrucaria) The mixed fermentation liquor comprises: the method comprises the following steps of (1) mixing a phanerochaete chrysosporium seed solution and a myrothecium verrucaria seed solution in a ratio of 1:1, stirring for 3-5 minutes, inoculating into a fermentation culture medium according to the inoculation amount of 3-6% of the volume ratio, and culturing at 28 ℃ for 5 days to obtain the phanerochaete chrysosporium-lachnum verrucosum mixed fermentation liquor.
The bacillus licheniformis-pseudomonas fluorescens mixed fermentation liquorThe method comprises the following steps: the effective viable count of Bacillus licheniformis is 2.2-3.0 × 1010cfu•L−1 The effective viable count of the pseudomonas fluorescens is 1.0-1.6 multiplied by 1010cfu·L−1(ii) a The effective viable count of the streptomyces fradiae in the streptomyces fradiae fermentation liquor is 1.4-3.3 multiplied by 106cfu·L−1(ii) a In the Phanerochaete chrysosporium-Myrothecium verrucaria mixed fermentation broth: the effective viable count of Phanerochaete chrysosporium is 0.8-2.5 × 108cfu·L−1The effective viable count of Myrothecium verrucaria is 2.6-5.0 × 108cfu·L−1(ii) a The total number of effective viable bacteria in the mixed solution is 3.12-5.50 multiplied by 109cfu·L−1。
The Bacillus licheniformis (A), (B)Bacillus licheniformis) Is ACCC 11080; pseudomonas fluorescens (Pseudomonas fluorescens) Is ACCC 10646; streptomyces fradiae (Streptomyces fradiae) Is ACCC 40124; phanerochaete chrysosporium (A)Phanerochaete chrysosporium Burdsall) Is ACCC 30530; myrothecium verrucaria (A), (B), (C)Myrothecium verrucaria) Is ACCC 30197.
And uniformly mixing the mixed solution and the chenopodium quinoa peel, drying at 35 ℃ for 48h, and crushing by 200 meshes to obtain the biocontrol microbial inoculum for preventing and treating root-knot nematodes.
Preferably: the Bacillus licheniformis (A), (B)Bacillus licheniformis) Pseudomonas fluorescens (Pseudomonas fluorescens) The mixed fermentation liquor comprises: the bacillus licheniformis seed solution and the pseudomonas fluorescens seed solution are prepared according to the following steps of: 1, stirring for 3-5min, inoculating to a fermentation culture medium according to the inoculum size of 6% in volume ratio, and culturing at 30 ℃ for 18-22h to obtain the product;
said Streptomyces fradiae (A), (B), (CStreptomyces fradiae) The fermentation liquor is: inoculating the streptomyces fradiae seed liquid into a fermentation culture medium according to the inoculation amount of 8% in volume ratio, and culturing for 7d at 29 ℃ to obtain streptomyces fradiae fermentation liquid;
phanerochaete chrysosporium (A)Phanerochaete chrysosporium Burdsall) Myrothecium verrucaria (A. japonicas (A.)), (B. verrucosa (B.))Myrothecium verrucaria) The mixed fermentation liquor comprises: mixing Phanerochaete chrysosporium seed liquid and Myrothecium verrucaria seed liquid in the volume ratio of 1 to 1, stirring for 3-5min, and inoculating according to the inoculation amount of 5% in volume ratioCulturing in fermentation culture medium at 28 deg.C for 5d to obtain mixed fermentation liquid of Phanerochaete chrysosporium and Myrothecium verrucaria.
The Bacillus licheniformis (A), (B)Bacillus licheniformis) Pseudomonas fluorescens (Pseudomonas fluorescens) The fermentation medium adopted in the mixed fermentation liquid is as follows: corn starch 15 g.L−1Molasses 10 g.L−1Peptone 1 g. L−1Glucose 2 g.L−1Ammonium sulfate 5 g.L−1，KH2PO4 0.45 g·L−1，MgSO4 0.45 g·L−1；
Said Streptomyces fradiae (A), (B), (CStreptomyces fradiae) The fermentation medium adopted in the fermentation liquid is as follows: glucose 6.0 g.L−1Soybean cake powder 6.0 g.L−1Soluble starch 35 g.L−1Magnesium sulfate 2 g.L−1Calcium carbonate 2 g.L−1；
Phanerochaete chrysosporium (A)Phanerochaete chrysosporium Burdsall) Myrothecium verrucaria (A. japonicas (A.)), (B. verrucosa (B.))Myrothecium verrucaria) The fermentation medium adopted in the mixed fermentation liquid is as follows: potato 200 g.L−1Soluble starch 3 g. L−1Sodium nitrate 0.8 g.L−1Lactose 15 g.L−1Beef extract 5 g.L−13 g.L ammonium sulfate−1。
The application method of the prepared biocontrol microbial inoculum in field nematode killing is hole application, 8-12g of hole application is carried out around each root line of the vegetables in the protected area, and the dosage per mu is 12-25 kg.
It should be noted that the seed culture and fermentor culture methods of the present invention are conventional in the art and are not described in detail.
The invention selects five strains from various biocontrol bacteria, wherein: the fungi Phanerochaete chrysosporium and Myrothecium verrucaria kill root-knot nematodes mainly through conidium parasitism eggs and a J2 mode; metabolites of bacteria bacillus licheniformis and pseudomonas fluorescens kill root-knot nematodes J2, promote root development of plants and improve disease resistance of plants; the metabolite of actinomycete streptomyces fradiae kills root-knot nematode J2, and simultaneously promotes the number of soil fungi, so as to cause the competition of nutrition and space with the root-knot nematode. The five strains are not antagonistic to each other and are synergistic to each other for disinsection, and the preparation process is relatively simple and the application prospect is wide. The invention can prevent and control vegetable root knot nematode disease, promote plant growth and development and improve plant disease resistance.
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FIG. 1 shows the field control effect of the biocontrol microbial inoculum for controlling root-knot nematodes of melons, which is prepared by the invention, and in the figure: a-b is the root system of the melon which is not controlled; c-d, treating and preventing the root system of the muskmelon.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present application, the technical solutions in the present application will be described below clearly and completely in conjunction with the specific embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1: a biocontrol microbial inoculum for preventing and treating root-knot nematode is prepared from Bacillus licheniformis (Bacillus licheniformis)Bacillus licheniformis) Pseudomonas fluorescens (Pseudomonas fluorescens) Mixed fermentation broth, Streptomyces fradiae: (Streptomyces fradiae) Fermentation broth, Phanerochaete chrysosporium: (Phanerochaete chrysosporium Burdsall) Myrothecium verrucaria (A. japonicas (A.)), (B. verrucosa (B.))Myrothecium verrucaria) The mixed fermentation liquor is prepared according to the following steps of 2: 2: 6, stirring for 10 minutes to obtain a liquid microbial inoculum; the liquid microbial inoculum and the quinoa peel are uniformly mixed according to the proportion of 1kg to 6kg, dried at 35 ℃ for 48h and crushed by 200 meshes to obtain the biocontrol microbial inoculum for preventing and treating the root-knot nematodes.
Wherein: bacillus licheniformis (Bacillus licheniformis) Pseudomonas fluorescens (Pseudomonas fluorescens) The mixed fermentation liquor comprises: the bacillus licheniformis seed solution and the pseudomonas fluorescens seed solution are prepared according to the following steps of: 1, stirring for 3-5min, inoculating to fermentation medium at 6% volume ratio, and culturing at 30 deg.C for 18-22h to obtainA bacillus licheniformis-pseudomonas fluorescens mixed fermentation liquor; the fermentation medium is as follows: corn starch 15 g.L−1Molasses 10 g.L−1Peptone 1 g. L−1Glucose 2 g.L−1Ammonium sulfate 5 g.L−1，KH2PO4 0.45 g·L−1，MgSO4 0.45 g·L−1；
Streptomyces fradiae (Streptomyces fradiae) The fermentation liquor is: inoculating the Streptomyces fradiae seed liquid into a fermentation medium according to the inoculation amount of 8% by volume ratio, and culturing for 7d at 29 ℃ to obtain a Streptomyces fradiae fermentation liquid; the fermentation medium is as follows: glucose 6.0 g.L−1Soybean cake powder 6.0 g.L−1Soluble starch 35 g.L−1Magnesium sulfate 2 g.L−1Calcium carbonate 2 g.L−1；
Phanerochaete chrysosporium (A)Phanerochaete chrysosporium Burdsall) Myrothecium verrucaria (A. japonicas (A.)), (B. verrucosa (B.))Myrothecium verrucaria) The mixed fermentation liquor comprises: the method comprises the following steps of (1) mixing a phanerochaete chrysosporium seed solution and a myrothecium verrucaria seed solution in a ratio of 1:1, stirring for 3-5 minutes, inoculating into a fermentation culture medium according to the inoculation amount of 5% of the volume ratio, and culturing at 28 ℃ for 5 days to obtain the phanerochaete chrysosporium-lachnum wartii mixed fermentation liquor. The fermentation medium is as follows: potato 200 g.L−1Soluble starch 3 g. L−1Sodium nitrate 0.8 g.L−1Lactose 15 g.L−1Beef extract 5 g.L−13 g.L ammonium sulfate−1。
In the mixed fermentation liquor of the bacillus licheniformis and the pseudomonas fluorescens: the effective viable count of Bacillus licheniformis is 2.2-3.0 × 1010cfu•L−1 The effective viable count of the pseudomonas fluorescens is 1.0-1.6 multiplied by 1010cfu·L−1(ii) a The effective viable count of the streptomyces fradiae in the streptomyces fradiae fermentation liquor is 1.4-3.3 multiplied by 106cfu·L−1(ii) a In the Phanerochaete chrysosporium-Myrothecium verrucaria mixed fermentation broth: the effective viable count of Phanerochaete chrysosporium is 0.8-2.5 × 108cfu·L−1The effective viable count of Myrothecium verrucaria is 2.6-5.0 × 108cfu·L−1(ii) a In the mixed solution is effectiveThe total number of viable bacteria is 3.12-5.50 × 109cfu·L−1。
The Bacillus licheniformis (A), (B)Bacillus licheniformis) Is ACCC 11080; pseudomonas fluorescens (Pseudomonas fluorescens) Is ACCC 10646; streptomyces fradiae (Streptomyces fradiae) Is ACCC 40124; phanerochaete chrysosporium (A)Phanerochaete chrysosporium Burdsall) Is ACCC 30530; myrothecium verrucaria (A), (B), (C)Myrothecium verrucaria) Is ACCC 30197.
The application method of the prepared biocontrol microbial inoculum in field nematode killing is hole application, 8g of the biocontrol microbial inoculum is hole applied around each root line of the vegetables in the protected field, and the dosage of the biocontrol microbial inoculum per mu is 12 kg.
Example 2: a biocontrol microbial inoculum for preventing and treating root-knot nematode is prepared from Bacillus licheniformis (Bacillus licheniformis)Bacillus licheniformis) Pseudomonas fluorescens (Pseudomonas fluorescens) Mixed fermentation broth, Streptomyces fradiae: (Streptomyces fradiae) Fermentation broth, Phanerochaete chrysosporium: (Phanerochaete chrysosporium Burdsall) Myrothecium verrucaria (A. japonicas (A.)), (B. verrucosa (B.))Myrothecium verrucaria) The mixed fermentation liquor is prepared according to the following steps of 3: 1: 7, stirring for 10 minutes to obtain a liquid microbial inoculum; the liquid microbial inoculum and the quinoa peel are uniformly mixed according to the proportion of 1kg to 8kg, dried at 35 ℃ for 48h and crushed by 200 meshes to obtain the biocontrol microbial inoculum for preventing and treating the root-knot nematodes.
Wherein: bacillus licheniformis (Bacillus licheniformis) Pseudomonas fluorescens (Pseudomonas fluorescens) The mixed fermentation liquor comprises: the bacillus licheniformis seed solution and the pseudomonas fluorescens seed solution are mixed according to the proportion of 1:1, stirring for 3-5 minutes, inoculating into a fermentation culture medium according to the inoculation amount of 3% of the volume ratio, and culturing at 30 ℃ for 18-22 hours to obtain a bacillus licheniformis-pseudomonas fluorescens mixed fermentation liquor; the fermentation medium is as follows: corn starch 15 g.L−1Molasses 10 g.L−1Peptone 1 g. L−1Glucose 2 g.L−1Ammonium sulfate 5 g.L−1，KH2PO4 0.45 g·L−1，MgSO4 0.45 g·L−1；
Streptomyces fradiae (Streptomyces fradiae) The fermentation liquor is: inoculating the streptomyces fradiae seed liquid into a fermentation culture medium according to the inoculation amount of 5% by volume ratio, and culturing for 7d at 29 ℃ to obtain streptomyces fradiae fermentation liquid; the fermentation medium is as follows: glucose 6.0 g.L−1Soybean cake powder 6.0 g.L−1Soluble starch 35 g.L−1Magnesium sulfate 2 g.L−1Calcium carbonate 2 g.L−1；
Phanerochaete chrysosporium (A)Phanerochaete chrysosporium Burdsall) Myrothecium verrucaria (A. japonicas (A.)), (B. verrucosa (B.))Myrothecium verrucaria) The mixed fermentation liquor comprises: the method comprises the following steps of (1) mixing a phanerochaete chrysosporium seed solution and a myrothecium verrucaria seed solution in a ratio of 1:1, stirring for 3-5 minutes, inoculating into a fermentation culture medium according to the inoculation amount of 3% of the volume ratio, and culturing at 28 ℃ for 5 days to obtain the phanerochaete chrysosporium-lachnum wartii mixed fermentation liquor. The fermentation medium is as follows: potato 200 g.L−1Soluble starch 3 g. L−1Sodium nitrate 0.8 g.L−1Lactose 15 g.L−1Beef extract 5 g.L−13 g.L ammonium sulfate−1。
In the mixed fermentation liquor of the bacillus licheniformis and the pseudomonas fluorescens: the effective viable count of Bacillus licheniformis is 2.2-3.0 × 1010cfu•L−1 The effective viable count of the pseudomonas fluorescens is 1.0-1.6 multiplied by 1010cfu·L−1(ii) a The effective viable count of the streptomyces fradiae in the streptomyces fradiae fermentation liquor is 1.4-3.3 multiplied by 106cfu·L−1(ii) a In the Phanerochaete chrysosporium-Myrothecium verrucaria mixed fermentation broth: the effective viable count of Phanerochaete chrysosporium is 0.8-2.5 × 108cfu·L−1The effective viable count of Myrothecium verrucaria is 2.6-5.0 × 108cfu·L−1(ii) a The total number of effective viable bacteria in the mixed solution is 3.12-5.50 multiplied by 109cfu·L−1。
The Bacillus licheniformis (A), (B)Bacillus licheniformis) Is ACCC 11080; pseudomonas fluorescens (Pseudomonas fluorescens) Is ACCC 10646; streptomyces fradiae (Streptomyces fradiae) Is ACCC 40124; phanerochaete chrysosporium (A)Phanerochaete chrysosporium Burdsall) Is ACCC 30530; myrothecium verrucaria (A), (B), (C)Myrothecium verrucaria) Is ACCC 30197.
The application method of the prepared biocontrol microbial inoculum in field nematode killing is hole application, 10g of the biocontrol microbial inoculum is hole applied around each root line of the vegetables in the protected field, and the dosage of the biocontrol microbial inoculum per mu is 20 kg.
Example 3: a biocontrol microbial inoculum for preventing and treating root-knot nematode is prepared from Bacillus licheniformis (Bacillus licheniformis)Bacillus licheniformis) Pseudomonas fluorescens (Pseudomonas fluorescens) Mixed fermentation broth, Streptomyces fradiae: (Streptomyces fradiae) Fermentation broth, Phanerochaete chrysosporium: (Phanerochaete chrysosporium Burdsall) Myrothecium verrucaria (A. japonicas (A.)), (B. verrucosa (B.))Myrothecium verrucaria) The mixed fermentation liquor is prepared according to the following steps of 5: 3: 1, stirring for 10 minutes to obtain a liquid microbial inoculum; the liquid microbial inoculum and the quinoa peel are uniformly mixed according to the proportion of 1kg to 7kg, dried at 35 ℃ for 48h and crushed by 200 meshes to obtain the biocontrol microbial inoculum for preventing and treating the root-knot nematodes.
Wherein: bacillus licheniformis (Bacillus licheniformis) Pseudomonas fluorescens (Pseudomonas fluorescens) The mixed fermentation liquor comprises: the bacillus licheniformis seed solution and the pseudomonas fluorescens seed solution are mixed according to the weight ratio of 1.5: 1, stirring for 3-5 minutes, inoculating into a fermentation culture medium according to the inoculation amount of 8% of the volume ratio, and culturing at 30 ℃ for 18-22 hours to obtain a bacillus licheniformis-pseudomonas fluorescens mixed fermentation liquor; the fermentation medium is as follows: corn starch 15 g.L−1Molasses 10 g.L−1Peptone 1 g. L−1Glucose 2 g.L−1Ammonium sulfate 5 g.L−1，KH2PO4 0.45 g·L−1，MgSO4 0.45 g·L−1；
Streptomyces fradiae (Streptomyces fradiae) The fermentation liquor is: inoculating the streptomyces fradiae seed liquid into a fermentation culture medium according to the inoculation amount of 3% by volume ratio, and culturing for 7d at 29 ℃ to obtain streptomyces fradiae fermentation liquid; the fermentation medium is as follows: glucose 6.0 g.L−1Soybean cake powder 6.0 g.L−1Soluble starch 35 g.L−1Magnesium sulfate 2 g.L−1Calcium carbonate 2 g.L−1；
Phanerochaete chrysosporium (A)Phanerochaete chrysosporium Burdsall) Myrothecium verrucaria (A. japonicas (A.)), (B. verrucosa (B.))Myrothecium verrucaria) The mixed fermentation liquor comprises: the method comprises the following steps of (1) mixing a phanerochaete chrysosporium seed solution and a myrothecium verrucaria seed solution in a ratio of 1:1, stirring for 3-5 minutes, inoculating into a fermentation culture medium according to the inoculation amount of 6% of the volume ratio, and culturing at 28 ℃ for 5 days to obtain the phanerochaete chrysosporium-lachnum wartii mixed fermentation liquor. The fermentation medium is as follows: potato 200 g.L−1Soluble starch 3 g. L−1Sodium nitrate 0.8 g.L−1Lactose 15 g.L−1Beef extract 5 g.L−13 g.L ammonium sulfate−1。
In the mixed fermentation liquor of the bacillus licheniformis and the pseudomonas fluorescens: the effective viable count of Bacillus licheniformis is 2.2-3.0 × 1010cfu•L−1 The effective viable count of the pseudomonas fluorescens is 1.0-1.6 multiplied by 1010cfu·L−1(ii) a The effective viable count of the streptomyces fradiae in the streptomyces fradiae fermentation liquor is 1.4-3.3 multiplied by 106cfu·L−1(ii) a In the Phanerochaete chrysosporium-Myrothecium verrucaria mixed fermentation broth: the effective viable count of Phanerochaete chrysosporium is 0.8-2.5 × 108cfu·L−1The effective viable count of Myrothecium verrucaria is 2.6-5.0 × 108cfu·L−1(ii) a The total number of effective viable bacteria in the mixed solution is 3.12-5.50 multiplied by 109cfu·L−1。
The Bacillus licheniformis (A), (B)Bacillus licheniformis) Is ACCC 11080; pseudomonas fluorescens (Pseudomonas fluorescens) Is ACCC 10646; streptomyces fradiae (Streptomyces fradiae) Is ACCC 40124; phanerochaete chrysosporium (A)Phanerochaete chrysosporium Burdsall) Is ACCC 30530; myrothecium verrucaria (A), (B), (C)Myrothecium verrucaria) Is ACCC 30197.
The application method of the prepared biocontrol microbial inoculum in field nematode killing is hole application, 12g of hole application is carried out around each root line of the vegetables in the protected area, and the dosage per mu is 25 kg.
Experimental example 1: and (3) field test: the experiment was carried out on vegetable fields where the pennisetum farinaceus developed severe root-knot nematodes. The tested crop is melon, and the microbial inoculum is applied before transplanting. The dosage of each strain is 12g of the complex microbial inoculum, and the contrast is the equivalent quinoa wheat bran. Root knot indexes are investigated in the harvesting period, and the prevention and treatment effects are evaluated. The severity of root nodule is 0-10 grade, and the preventing and treating effect = (disease index of control area-disease index of treatment area)/disease index of control area x 100.
The control effect is shown in table 1, the result shows that the compound microbial inoculum can effectively control the root knot nematode disease of the melons, the control effect is 53.7%, the biological yield of plants is obviously improved, the plants in the treatment area are increased by 25.3%, the fresh weight of roots is increased by 50.9%, the root system of the melons in the treatment area is developed, the root system of the melons in the test control area is fine, and only a small amount of root knots are formed on the melons in the treatment area as shown in the figure 1.
TABLE 1 field control effect of the composition on meloidogyne
Figure 270672DEST_PATH_IMAGE002

Claims (4)

1. A biocontrol microbial inoculum for preventing and treating root-knot nematodes is characterized in that: the preparation method comprises the following steps of mixing Bacillus licheniformis (Bacillus licheniformis) -Pseudomonas fluorescens (Pseudomonas fluorescens) mixed fermentation liquor, Streptomyces fradiae (Streptomyces fradiae) fermentation liquor, Phanerochaete chrysosporium (Phanerochaete chrysosporium Burdsall) -Myrothecium verrucaria (Myrothecium verrucaria) mixed fermentation liquor according to the volume ratio of 2-5: 1-3: 1-7, and uniformly mixing the mixed liquor and quinoa peel according to the mass ratio of 1kg:6-8 kg;
wherein: the mixed fermentation liquor of Bacillus licheniformis (Bacillus licheniformis) -Pseudomonas fluorescens (Pseudomonas fluorescens) is as follows: mixing the bacillus licheniformis seed solution and the pseudomonas fluorescens seed solution according to the volume ratio of 1-2: 1, stirring for 3-5 minutes, inoculating the mixture into a fermentation culture medium according to the inoculation amount of 3-8% of the volume ratio, and culturing at 30 ℃ for 18-22 hours to obtain the bacillus licheniformis-pseudomonas fluorescens mixed fermentation liquor;
the fermentation broth of Streptomyces fradiae is: inoculating the Streptomyces fradiae seed liquid into a fermentation culture medium according to the inoculation amount of 3-8% in volume ratio, and culturing for 7d at 29 ℃ to obtain a Streptomyces fradiae fermentation liquid;
phanerochaete chrysosporium (Phanerochaete chrysosporium Burdsall) -Myrothecium verrucaria mixed fermentation broth is: mixing the phanerochaete chrysosporium seed liquid and the myrothecium verrucaria seed liquid according to the volume ratio of 1:1, stirring for 3-5 minutes, inoculating the mixture into a fermentation culture medium according to the inoculation amount of 3-6% of the volume ratio, and culturing for 5 days at 28 ℃ to obtain a phanerochaete chrysosporium-myrothecium verrucaria mixed fermentation liquid;
in the mixed fermentation liquor of the bacillus licheniformis and the pseudomonas fluorescens: the effective viable count of Bacillus licheniformis is 2.2-3.0 × 1010cfu·L-1The effective viable count of the pseudomonas fluorescens is 1.0-1.6 multiplied by 1010cfu·L-1(ii) a The effective viable count of the streptomyces fradiae in the streptomyces fradiae fermentation liquor is 1.4-3.3 multiplied by 106cfu·L-1(ii) a In the Phanerochaete chrysosporium-Myrothecium verrucaria mixed fermentation broth: the effective viable count of Phanerochaete chrysosporium is 0.8-2.5 × 108cfu·L-1The effective viable count of Myrothecium verrucaria is 2.6-5.0 × 108cfu·L-1(ii) a The total number of effective viable bacteria in the mixed solution is 3.12-5.50 multiplied by 109cfu·L-1；
In the fermentation liquor: the Bacillus licheniformis (Bacillus licheniformis) is ACCC 11080; pseudomonas fluorescens (Pseudomonas fluorescens) is ACCC 10646; streptomyces fradiae (Streptomyces fradiae) is ACCC 40124; phanerochaete chrysosporium (Phanerochaete chrysosporium Burdsall) is ACCC 30530; myrothecium verrucaria (Myrothecium verrucaria) ACCC 30197;
the fermentation medium adopted in the mixed fermentation liquid of the Bacillus licheniformis (Bacillus licheniformis) -Pseudomonas fluorescens (Pseudomonas fluorescens) is as follows: corn starch 15 g.L-1Molasses 10 g.L-1Peptone 1 g. L-1Glucose 2 g.L-1Ammonium sulfate 5 g.L-1，KH2PO40.45g·L-1，MgSO40.45g·L-1；
The fermentation medium adopted in the Streptomyces fradiae fermentation liquid is as follows: glucose 6.0 g.L-1Soybean cake powder 6.0 g.L-1Soluble starch 35 g.L-1Magnesium sulfate 2 g.L-1Calcium carbonate 2 g.L-1；
The fermentation culture medium adopted in the mixed fermentation liquor of Phanerochaete chrysosporium (Phanerochaete chrysosporium Burdsall) -Myrothecium verrucaria (Myrothecium verrucaria) is as follows: potato 200 g.L-1Soluble starch 3 g. L-1Sodium nitrate 0.8 g.L-1Lactose 15 g.L-1Beef extract 5 g.L-13 g.L ammonium sulfate-1。
2. The biocontrol microbial inoculum for controlling root-knot nematodes of claim 1, which is characterized in that: and uniformly mixing the mixed solution and the chenopodium quinoa peel, drying at 35 ℃ for 48h, and crushing by 200 meshes to obtain the biocontrol microbial inoculum for preventing and treating root-knot nematodes.
3. The biocontrol microbial inoculum for controlling root-knot nematodes of claim 1, which is characterized in that: the mixed fermentation liquor of the Bacillus licheniformis (Bacillus licheniformis) -Pseudomonas fluorescens (Pseudomonas fluorescens) is as follows: mixing the Bacillus licheniformis seed solution and the Pseudomonas fluorescens seed solution in a volume ratio of 2: 1, stirring for 3-5min, inoculating the mixture into a fermentation culture medium according to an inoculum size of 6% in volume ratio, and culturing at 30 ℃ for 18-22h to obtain the bacillus licheniformis seed solution;
the fermentation liquor of the Streptomyces fradiae is as follows: inoculating the streptomyces fradiae seed liquid into a fermentation culture medium according to the inoculation amount of 8% in volume ratio, and culturing for 7d at 29 ℃ to obtain streptomyces fradiae fermentation liquid;
the Phanerochaete chrysosporium (Phanerochaete chrysosporium Burdsall) -Myrothecium verrucaria mixed fermentation broth is as follows: mixing the phanerochaete chrysosporium seed liquid and the Myrothecium verrucaria seed liquid according to the volume ratio of 1:1, stirring for 3-5 minutes, inoculating into a fermentation culture medium according to the inoculation amount of 5% of the volume ratio, and culturing for 5 days at 28 ℃ to obtain the phanerochaete chrysosporium-Myrothecium verrucaria mixed fermentation liquid.
4. The biocontrol microbial inoculum for controlling root-knot nematodes of claim 1, which is characterized in that: the application method of the prepared biocontrol microbial inoculum in field nematode killing is hole application, 8-12g of hole application is carried out around each root line of the vegetables in the protected area, and the dosage per mu is 12-25 kg.
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108395998A (en) * 2017-02-08 2018-08-14 山东中创亿丰肥料集团有限公司 A kind of desinsection biological compound fungi agent and preparation method thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
"疣孢漆斑菌(Myrothecium verrucaria)对蔬菜根结线虫寄生致死作用及应用研究";董海龙;《中国博士学位论文全文数据库 农业科技辑（月刊 ）》;20160715(第 07 期);D046-5 *
"黄孢原毛平革菌对黄瓜连作土壤酚酸物质的降解";徐淑霞 等;《应用生态学报》;20081130;第19卷(第11期);第2480-2484页 *

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