Biocontrol bacterium composition and application of biocontrol bacterium composition in prevention and treatment of melanogaster

Abstract

The invention relates to the technical field of prevention and control of melanogaster. The invention provides a biocontrol bacterium composition and application of the biocontrol bacterium composition in prevention and treatment of melanogaster. The biocontrol bacterium composition comprises the following raw materials in parts by weight: 45-55 parts of a metarhizium anisopliae bacterium agent, 15-25 parts of a bacillus thuringiensis bacterium agent, 10-15 parts of a bacillus licheniformis bacterium agent and 10-15 parts of a bacillus laterosporus bacterium agent. The biocontrol bacterium composition disclosed by the invention contains a plurality of different strains, and the different strains are matched with one another, so that the melanogaster can be effectively prevented and treated, and the killing efficiency of the product is improved. And the used strain is environment-friendly, the efficiency is higher after the strain is loaded by a carrier, and the strain is more convenient to use.

Description

Technical Field

[0001] The invention relates to the technical field of black fungus insect control, in particular to a biocontrol fungus composition and application thereof in controlling black fungus insect control. Background Art

[0002] Black fungus worm is a warehouse pest of the family Tenebrioidae, order Coleoptera, class Insecta. Its hosts include cotton, wood, jute, costusroot, rice, peanuts, grits, raw medicinal materials, dates, corn, cocoa beans, mung beans, sesame, cardamom, ebony wood, white crown, and betel nut. It harms grains and their processed products, dried fruits, cocoa, poria hair, coconut, cardamom, etc. It can also eat insect corpses, insect feces, and rotten inclusions, and its diet is very complex. It occurs most frequently in damp grains, old grains, grain stubs, dark and damp places with dust and debris in granaries and grain processing plants.

[0003] Black fungus worms often gather together with black mealworms, yellow mealworms, and small fungus worms in large numbers to cause harm. Adults or larvae often gather in dark corners of warehouses, under bricks and stones, wooden boards, in dust and debris, or in grain debris to hibernate. Adults are gregarious, phototactic, feign death, and cannibalistic. They can fly and crawl well, like to eat insect corpses and debris, can resist starvation for 11 to 24 days, and like to eat moist grains; the lifespan of adults can reach 1 year under hot and humid conditions. Larvae like to eat wet grains and flour, have feign death, are carnivorous, live in groups, and are phototactic, and can withstand starvation for 3 to 9 days. Summary of the invention

[0004] The purpose of the present invention is to provide a biocontrol fungus composition and its application in preventing and controlling black fungus insects, and to utilize microbial preparations to efficiently prevent and control black fungus insects, thereby effectively preventing and controlling black fungus insects.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The invention provides a biocontrol bacteria composition, which comprises the following raw materials in parts by weight: 45-55 parts of Metarhizium anisopliae agent, 15-25 parts of Bacillus thuringiensis agent, 10-15 parts of Bacillus licheniformis agent, and 10-15 parts of Bacillus laterosporus agent.

[0007] Preferably, the effective viable count of Metarhizium anisopliae in the Metarhizium anisopliae agent is 2.5 to 2.8×10 10 cfu / g;

[0008] The preparation method of the green muscardine agent is as follows: inoculating green muscardine into a PDA culture medium, culturing at 26-30° C., and drying to obtain the green muscardine agent;

[0009] The type of Metarhizium is Metarhizium anisopliae or Metarhizium umbellatum.

[0010] Preferably, the effective viable count of Bacillus thuringiensis in the Bacillus thuringiensis bacterial agent is 1.3 - 1.8×10 10 cfu / g;

[0011] The preparation method of the Bacillus thuringiensis bacterial agent is as follows: inoculate Bacillus thuringiensis into a Bacillus thuringiensis culture medium, culture it at 30 - 32°C, and obtain the Bacillus thuringiensis bacterial agent after drying;

[0012] The formula of the Bacillus thuringiensis culture medium is: 25 - 45 g / L of glucose, 1 - 2 g / L of yeast extract, 60 - 80 g / L of corn steep liquor, and the balance is water.

[0013] Preferably, the effective viable count of Bacillus licheniformis in the Bacillus licheniformis bacterial agent is 2.2 - 2.4×10 10 cfu / g;

[0014] The preparation method of the Bacillus licheniformis bacterial agent is as follows: inoculate Bacillus licheniformis into an LB medium, culture it at 28 - 30°C, and obtain the Bacillus licheniformis bacterial agent after drying.

[0015] Preferably, the effective viable count of Bacillus laterosporus in the Bacillus laterosporus bacterial agent is 1.8 - 2.2×10 10 cfu / g;

[0016] The preparation method of the Bacillus laterosporus bacterial agent is as follows: inoculate Bacillus laterosporus into an LB medium, culture it at 28 - 30°C, and obtain the Bacillus laterosporus bacterial agent after drying.

[0017] The present invention also provides the application of the biocontrol bacterial composition in controlling Alphitobius diaperinus.

[0018] The present invention also provides the application of the biocontrol bacterial composition in the preparation of a product for controlling Alphitobius diaperinus.

[0019] The present invention also provides a product for controlling Alphitobius diaperinus, and the product comprises the biocontrol bacterial composition and a carrier.

[0020] Preferably, the carrier is wheat bran, rice husk or peanut shell.

[0021] The present invention also provides a preparation method of the product, which comprises the following steps: crush the carrier and adjust the water content to 45 - 65%, inoculate the biocontrol bacterial composition, mix and culture at 28 - 37°C for 24 - 60 h, and obtain the product after drying;

[0022] The particle size of the crushed carrier ≤ 5 mm;

[0023] The inoculation amount of the biocontrol bacterial composition is 0.8 - 1.2% of the mass of the carrier;

[0024] The water content of the dried product is 8-12%.

[0025] The present invention provides a biocontrol bacteria composition and its application in controlling Alphitobius diaperinus. The biocontrol bacteria composition comprises the following raw materials in parts by weight: 45-55 parts of Metarhizium anisopliae bacterial agent, 15-25 parts of Bacillus thuringiensis bacterial agent, 10-15 parts of Bacillus licheniformis bacterial agent, and 10-15 parts of Bacillus laterosporus bacterial agent. The biocontrol bacteria composition of the present invention contains a variety of different bacterial species, and the different bacterial species cooperate with each other, which can effectively control Alphitobius diaperinus and improve the killing efficiency of the product. Moreover, the used bacterial species are environmentally friendly, and the efficiency is exerted faster after being loaded on a carrier, which is more convenient to use. Detailed implementation mode

[0026] The present invention provides a biocontrol bacteria composition, which comprises the following raw materials in parts by weight: 45-55 parts of Metarhizium anisopliae bacterial agent, 15-25 parts of Bacillus thuringiensis bacterial agent, 10-15 parts of Bacillus licheniformis bacterial agent, and 10-15 parts of Bacillus laterosporus bacterial agent.

[0027] In the present invention, the number of parts of the Metarhizium anisopliae bacterial agent is preferably 48-52 parts, and more preferably 50 parts.

[0028] In the present invention, the number of parts of the Bacillus thuringiensis bacterial agent is preferably 18-22 parts, and more preferably 20 parts.

[0029] In the present invention, the number of parts of the Bacillus licheniformis bacterial agent is preferably 11-14 parts, and more preferably 12-13 parts.

[0030] In the present invention, the number of parts of the Bacillus laterosporus bacterial agent is preferably 11-14 parts, and more preferably 12-13 parts.

[0031] In the present invention, the effective viable count of Metarhizium anisopliae in the Metarhizium anisopliae bacterial agent is preferably 2.5-2.8×10 10 cfu / g, and more preferably 2.6-2.7×10 10 cfu / g.

[0032] In the present invention, the preparation method of the Metarhizium anisopliae bacterial agent is preferably: inoculating Metarhizium anisopliae into a PDA medium, culturing at 26-30°C, and drying to obtain the Metarhizium anisopliae bacterial agent. More preferably: inoculating Metarhizium anisopliae into a PDA medium, culturing at 28°C, and drying to obtain the Metarhizium anisopliae bacterial agent.

[0033] In the present invention, the species of Metarhizium anisopliae is preferably Metarhizium anisopliae var. acridum or Metarhizium anisopliae var. flavoviride.

[0034] In the present invention, the effective viable count of Bacillus thuringiensis in the Bacillus thuringiensis agent is preferably 1.3 - 1.8×10 10 cfu / g, and more preferably 1.5×10 10 cfu / g.

[0035] In the present invention, the preparation method of the Bacillus thuringiensis agent is preferably: inoculating Bacillus thuringiensis into a Bacillus thuringiensis medium, culturing at 30 - 32°C, and drying to obtain the Bacillus thuringiensis agent. More preferably: inoculating Bacillus thuringiensis into a Bacillus thuringiensis medium, culturing at 31°C, and drying to obtain the Bacillus thuringiensis agent.

[0036] In the present invention, the formula of the Bacillus thuringiensis medium is preferably: glucose 25 - 45 g / L, yeast extract 1 - 2 g / L, corn steep liquor 60 - 80 g / L, with the balance being water. More preferably: glucose 35 g / L, yeast extract 1.5 g / L, corn steep liquor 70 g / L, with the balance being water.

[0037] In the present invention, the effective viable count of Bacillus licheniformis in the Bacillus licheniformis agent is preferably 2.2 - 2.4×10 10 cfu / g, and more preferably 2.0×10 10 cfu / g.

[0038] In the present invention, the preparation method of the Bacillus licheniformis agent is preferably: inoculating Bacillus licheniformis into an LB medium, culturing at 28 - 30°C, and drying to obtain the Bacillus licheniformis agent. More preferably: inoculating Bacillus licheniformis into an LB medium, culturing at 29°C, and drying to obtain the Bacillus licheniformis agent.

[0039] In the present invention, the effective viable count of Bacillus laterosporus in the Bacillus laterosporus agent is preferably 1.8 - 2.2×10 10 cfu / g, and more preferably 2.0×10 10 cfu / g.

[0040] In the present invention, the preparation method of the Bacillus laterosporus agent is preferably: inoculating Bacillus laterosporus into an LB medium, culturing at 28 - 30°C, and drying to obtain the Bacillus laterosporus agent. More preferably: inoculating Bacillus laterosporus into an LB medium, culturing at 29°C, and drying to obtain the Bacillus laterosporus agent.

[0041] The present invention also provides the application of the biocontrol bacteria composition in controlling Alphitophagus bifasciatus.

[0042] The present invention also provides the application of the biocontrol bacteria composition in the preparation of a product for controlling Alphitophagus bifasciatus.

[0043] The present invention also provides a product for controlling Alphitobius diaperinus, and the product comprises the biocontrol bacterial composition and a carrier.

[0044] In the present invention, the carrier is preferably wheat bran, rice husk or peanut shell.

[0045] The present invention also provides a preparation method of the product, comprising the following steps: pulverizing the carrier, adjusting the water content to 45-65%, inoculating the biocontrol bacterial composition, culturing at 28-37°C for 24-60 h after mixing, and drying to obtain the product.

[0046] In the present invention, the adjusted water content is preferably 55%.

[0047] In the present invention, the culturing temperature is preferably 32-35°C.

[0048] In the present invention, the culturing time is preferably 36-48 h.

[0049] In the present invention, the particle size of the pulverized carrier is preferably ≤5 mm.

[0050] In the present invention, the inoculation amount of the biocontrol bacterial composition is preferably 0.8-1.2% of the mass of the carrier;

[0051] In the present invention, the water content of the dried product is preferably 8-12%.

[0052] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0053] Example 1

[0054] A preparation method of a biocontrol bacterial composition comprises the following steps:

[0055] (1) Inoculating Metarhizium anisopliae var. acridum into a PDA medium, culturing at 30°C, and drying to obtain a Metarhizium anisopliae var. acridum bacterial agent with an effective viable count of 2.5×10 10 cfu / g;

[0056] (2) Weighing raw materials according to the ratio of 45 g / L of glucose, 2 g / L of yeast extract, and 60 g / L of corn steep liquor, preparing a Bacillus thuringiensis medium, inoculating Bacillus thuringiensis into the Bacillus thuringiensis medium, culturing at 32°C, and drying to obtain a Bacillus thuringiensis bacterial agent with an effective viable count of 1.8×10 10 cfu / g;

[0057] (3) Inoculate Bacillus licheniformis into LB medium and culture it at 28 °C. After drying, a Bacillus licheniformis microbial agent with an effective viable count of 2.4×10 10 cfu / g is obtained;

[0058] (4) Inoculate Bacillus laterosporus into LB medium and culture it at 30 °C. After drying, a Bacillus laterosporus microbial agent with an effective viable count of 1.8×10 10 cfu / g is obtained;

[0059] (5) Mix the raw materials according to the mass ratio of 55 parts of Metarhizium anisopliae microbial agent, 25 parts of Bacillus thuringiensis microbial agent, 10 parts of Bacillus licheniformis microbial agent, and 10 parts of Bacillus laterosporus microbial agent to obtain the biocontrol bacterial composition.

[0060] Example 2

[0061] A preparation method of a biocontrol bacterial composition includes the following steps:

[0062] (1) Inoculate Metarhizium anisopliae var. flavoviride into PDA medium and culture it at 26 °C. After drying, a Metarhizium anisopliae microbial agent with an effective viable count of 2.8×10 10 cfu / g is obtained;

[0063] (2) Weigh the raw materials according to the ratio of 25 g / L of glucose, 1 g / L of yeast extract, and 80 g / L of corn steep liquor, and prepare a Bacillus thuringiensis medium. Inoculate Bacillus thuringiensis into the Bacillus thuringiensis medium and culture it at 30 °C. After drying, a Bacillus thuringiensis microbial agent with an effective viable count of 1.3×10 10 cfu / g is obtained;

[0064] (3) Inoculate Bacillus licheniformis into LB medium and culture it at 30 °C. After drying, a Bacillus licheniformis microbial agent with an effective viable count of 2.2×10 10 cfu / g is obtained;

[0065] (4) Inoculate Bacillus laterosporus into LB medium and culture it at 28 °C. After drying, a Bacillus laterosporus microbial agent with an effective viable count of 2.2×10 10 cfu / g is obtained;

[0066] (5) Mix the raw materials according to the mass ratio of 45 parts of Metarhizium anisopliae microbial agent, 15 parts of Bacillus thuringiensis microbial agent, 15 parts of Bacillus licheniformis microbial agent, and 15 parts of Bacillus laterosporus microbial agent to obtain the biocontrol bacterial composition.

[0067] Example 3

[0068] A preparation method of a biocontrol bacterial composition includes the following steps:

[0069] (1) Inoculate Metarhizium anisopliae var. anisopliae into PDA medium, culture it at 28 °C, and after drying, obtain a Metarhizium anisopliae var. anisopliae microbial inoculum with an effective viable count of 2.6×10 10 cfu / g;

[0070] (2) Weigh raw materials according to the ratio of 35 g / L of glucose, 1.5 g / L of yeast extract, and 70 g / L of corn steep liquor, prepare a Bacillus thuringiensis medium, inoculate Bacillus thuringiensis into the Bacillus thuringiensis medium, culture it at 31 °C, and after drying, obtain a Bacillus thuringiensis microbial inoculum with an effective viable count of 1.5×10 10 cfu / g;

[0071] (3) Inoculate Bacillus licheniformis into LB medium, culture it at 29 °C, and after drying, obtain a Bacillus licheniformis microbial inoculum with an effective viable count of 2.3×10 10 cfu / g;

[0072] (4) Inoculate Bacillus laterosporus into LB medium, culture it at 29 °C, and after drying, obtain a Bacillus laterosporus microbial inoculum with an effective viable count of 2.0×10 10 cfu / g;

[0073] (5) Mix the raw materials according to the mass ratio of 50 parts of Metarhizium anisopliae var. anisopliae microbial inoculum, 20 parts of Bacillus thuringiensis microbial inoculum, 13 parts of Bacillus licheniformis microbial inoculum, and 12 parts of Bacillus laterosporus microbial inoculum to obtain the biocontrol bacteria composition.

[0074] Example 4

[0075] A preparation method of a product for controlling Alphitophagus bifasciatus includes the following steps:

[0076] Crush wheat bran to a particle size ≤ 5 mm, adjust the water content to 45%, add the biocontrol bacteria composition prepared in Example 1 at 1.2% of the carrier mass, mix, culture at 37 °C for 24 h, and dry to a water content of 8% to obtain the product.

[0077] Example 5

[0078] A preparation method of a product for controlling Alphitophagus bifasciatus includes the following steps:

[0079] Crush rice husks to a particle size ≤ 5 mm, adjust the water content to 65%, add the biocontrol bacteria composition prepared in Example 2 at 0.8 - 1.2% of the carrier mass, mix, culture at 28 °C for 60 h, and dry to a water content of 12% to obtain the product.

[0080] Example 6

[0081] A preparation method of a product for controlling Alphitophagus bifasciatus includes the following steps:

[0082] Crush peanut shells to a particle size of ≤5 mm, adjust the water content to 55%, inoculate with the biocontrol bacterial composition prepared in Example 3 at 1% of the carrier mass, mix, and culture at 32 °C for 36 h, then dry to a water content of 10% to obtain the product.

[0083] Test Example

[0084] Take the products prepared in Examples 4 to 6 and aluminum phosphide insecticide for a comparative test, and each product is tested three times. The specific method is as follows:

[0085] Before the start of the test, select 1200 black fungus beetle larvae and 1200 adults. Group the larvae and adults, numbered as test groups 1 to 9 and control groups 1 to 3, with 100 larvae and 100 adults in each group. Use the products prepared in Examples 4 to 6 and aluminum phosphide insecticide for killing tests respectively. On the first day of the start of the test, feed the black fungus beetles in test groups 1 to 3 with the product prepared in Example 4, feed the black fungus beetles in test groups 4 to 6 with the product prepared in Example 5, feed the black fungus beetles in test groups 7 to 9 with the product prepared in Example 6, and fumigate the black fungus beetles in control groups 1 to 3 with aluminum phosphide insecticide.

[0086] After 7 days, count the death situation of black fungus beetles in each group, and the results are shown in Table 1.

[0087] Table 1

[0088]

[0089]

[0090] Conclusion: The biocontrol bacterial composition prepared by the present invention can effectively control black fungus beetles, and the killing effect is close to that of aluminum phosphide insecticide. At the same time, due to its environmental friendliness, the operation requirements are much lower than those of highly toxic aluminum phosphide insecticide.

[0091] From the above examples, it can be seen that the present invention provides a biocontrol bacterial composition and its application in controlling black fungus beetles. The biocontrol bacterial composition comprises the following raw materials in parts by weight: 45 - 55 parts of Metarhizium anisopliae bacterial agent, 15 - 25 parts of Bacillus thuringiensis bacterial agent, 10 - 15 parts of Bacillus licheniformis bacterial agent, and 10 - 15 parts of Bacillus laterosporus bacterial agent. The biocontrol bacterial composition of the present invention contains a variety of different bacterial species, and the different bacterial species cooperate with each other, which can effectively control black fungus beetles and improve the killing efficiency of the product. Moreover, the used bacterial species are environmentally friendly, and the efficiency is faster after being loaded on the carrier, which is more convenient to use.

[0092] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. A biocontrol bacteria composition, characterized in that: The biocontrol bacteria composition comprises the following raw materials in parts by weight: 45-55 parts of Metarhizium anisopliae agent, 15-25 parts of Bacillus thuringiensis agent, 10-15 parts of Bacillus licheniformis agent, and 10-15 parts of Bacillus laterosporus agent.

2. The composition according to claim 1, characterized in that The effective live bacteria count of Metarhizium anisopliae in the Metarhizium anisopliae inoculum is 2.5 to 2.8×10 10 cfu / g; The preparation method of the green muscardine agent is as follows: inoculating green muscardine into a PDA culture medium, culturing at 26-30° C., and drying to obtain the green muscardine agent; The type of Metarhizium is Metarhizium anisopliae or Metarhizium umbellatum.

3. The composition according to claim 1, characterized in that The effective live bacteria count of Bacillus thuringiensis in the Bacillus thuringiensis agent is 1.3 to 1.8×10 10 cfu / g; The preparation method of the Bacillus thuringiensis agent comprises: inoculating Bacillus thuringiensis into a Bacillus thuringiensis culture medium, culturing at 30-32° C., and drying to obtain the Bacillus thuringiensis agent; The formula of the Bacillus thuringiensis culture medium is: 25-45 g / L of glucose, 1-2 g / L of yeast extract, 60-80 g / L of corn steep liquor, and the balance is water.

4. The composition according to claim 1, characterized in that The effective live bacteria count of Bacillus licheniformis in the Bacillus licheniformis inoculant is 2.2 to 2.4×10 10 cfu / g; The preparation method of the Bacillus licheniformis agent is as follows: inoculating Bacillus licheniformis into LB culture medium, culturing at 28-30° C., and drying to obtain the Bacillus licheniformis agent.

5. The composition according to claim 1, characterized in that The effective viable bacteria count of Bacillus laterosporus in the Bacillus laterosporus inoculant is 1.8 to 2.2×10 10 cfu / g; The preparation method of the Bacillus laterosporus inoculant is as follows: inoculating the Bacillus laterosporus into an LB culture medium, culturing at 28-30° C., and drying to obtain the Bacillus laterosporus inoculant.

6. Use of the biocontrol fungus composition according to any one of claims 1 to 5 in controlling black fungus insects.

7. Use of the biocontrol fungus composition according to any one of claims 1 to 5 in the preparation of products for controlling black fungus insects.

8. A product for preventing and controlling black fungus insects, characterized in that: The product comprises the biocontrol bacteria composition according to any one of claims 1 to 5 and a carrier.

9. The product according to claim 8, characterized in that The carrier is wheat bran, rice husk or peanut husk.

10. The method for preparing the product according to claim 8 or 9, characterized in that: The method comprises the following steps: crushing the carrier and adjusting the water content to 45-65%, inoculating the biocontrol bacteria composition according to any one of claims 1-5, culturing at 28-37° C. for 24-60 hours after mixing, and drying to obtain the product; The particle size of the carrier after crushing is ≤5mm; The amount of the biocontrol bacteria composition introduced is 0.8-1.2% of the carrier mass; The moisture content of the dried product is 8-12%.

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