Bacillus amyloliquefaciens and metabolites thereof for use in stress resistance in vegetables

Abstract

The application discloses a bacillus zoogleoides BZ-32 and application of metabolites of the bacillus zoogleoides BZ-32 in vegetable stress resistance, and an effective component of the application is composed of bacillus zoogleoides fermentation liquor or metabolites, choline chloride and lentinan, wherein the weight percentage of the bacillus zoogleoides, choline chloride and lentinan is respectively 5-30% of bacillus zoogleoides metabolites, 5-30% of choline chloride and 0.2-5.0% of lentinan. The plant stress resistance composition can prevent and treat diseases such as root rot, anthracnose and powdery mildew of vegetables, and can prevent stress caused by excessively low temperature in winter and spring frost, and improve disease resistance and stress resistance of crops. The application can also be used for preventing and treating various corn diseases and strawberry diseases, and promoting growth and improving stress resistance.

Description

Technical Field

[0001] This invention relates to the field of pesticides, and in particular to compositions of Bacillus amyloliquefaciens and its metabolites for stress resistance in vegetables. This invention also relates to their uses. Background Technology

[0002] A strain of Bacillus velezensis BZ-32 was collected from soil in a greenhouse where tomatoes had been continuously cropped for many years.

[0003] Choline chloride is a type of vitamin B. Choline promotes fat metabolism in the liver and kidneys; it is also fundamental for the body's synthesis of acetylcholine, thus affecting nerve signal transmission. Additionally, choline is one of the methyl sources required for the synthesis of methionine in the body. Many foods contain natural choline, but the concentration is insufficient to meet the needs of modern feed industry for rapid animal growth. Therefore, synthetic choline should be added to feed to meet these needs. Choline deficiency can lead to fatty liver, slow growth, reduced egg production, and increased mortality. Choline chloride is also a photosynthetic promoter in plants, significantly increasing yield.

[0004] Shiitake mushrooms (Lentinus edodes) are basidiomycetes belonging to the Pleurotaceae family and contain a variety of effective medicinal components. In particular, lentinan (LNT) is a host immune enhancer with antiviral properties, regulating plant immune function, and stimulating interferon formation. The components of lentinan that enhance plant immunity are polysaccharides with a molecular weight exceeding 10,000. In my country, lentinan is currently used in medicine and for the prevention and control of plant viral diseases. Summary of the Invention

[0005] The purpose of this invention is to provide a composition of Bacillus amyloliquefaciens and its metabolites for the application of vegetable stress resistance. It consists of Bacillus amyloliquefaciens, choline chloride and lentinan, which can effectively prevent and control root rot, anthracnose, powdery mildew, gray mold and other diseases in vegetables. It can also prevent frost damage caused by low winter temperatures and late spring frosts, and improve the disease resistance and stress resistance of vegetables.

[0006] Based on the above objectives, the present invention provides a composition for applying Bacillus amyloliquefaciens and its metabolites to vegetable stress resistance, comprising an active ingredient and an adjuvant; wherein the active ingredient is composed of Bacillus amyloliquefaciens, choline chloride, and lentinan; the weight percentages of Bacillus amyloliquefaciens, choline chloride, and lentinan are 5-30% Bacillus amyloliquefaciens, 5-30% choline chloride, and 0.2-5.0% lentinan, respectively.

[0007] Optionally, the weight ratio of Bacillus amyloliquefaciens, choline chloride, and lentinan is 5~30:5~30:0.2~5.0.

[0008] Preferably, the weight ratio of Bacillus amyloliquefaciens, choline chloride and lentinan is 10-30:10-30:0.2-2.0.

[0009] Preferably, the weight ratio of Bacillus amyloliquefaciens, choline chloride, and lentinan is 30:20:0.5.

[0010] This invention can use wetting agents, fillers, and other additives to process the product into suspensions, wettable powders, or water-dispersible granules using traditional methods. The filler is selected from at least one of silica, diatomaceous earth, attapulgite, clay, and earthenware. The wetting agent is selected from at least one of sodium dodecyl sulfate, sodium lignosulfonate, and separating powder.

[0011] The present invention also provides the use of the above-mentioned pesticide composition, which can be used to prevent and control root rot, anthracnose and powdery mildew in vegetables, and can prevent frost damage caused by excessively low winter temperatures and late spring frosts, thereby improving the disease resistance and stress resistance of vegetables.

[0012] The method for using the above-mentioned pesticide composition to control the above-mentioned vegetable seedling diseases is as follows: at the early stage of vegetable disease, use Bacillus amyloliquefaciens•choline chloride•lentinan wettable powder, water-dispersible granules or suspension concentrate, add water to make a suspension and spray it.

[0013] The method for using the above-mentioned pesticide composition to defend against vegetable frost damage (or at the same time prevent and control vegetable seedling diseases) is as follows: Before winter and / or before the occurrence of late spring frost, use Bacillus amyloliquefaciens•choline chloride•lentinan wettable powder, water-dispersible granules or suspension concentrate, add water to make a suspension and spray it.

[0014] As can be seen from the above description and experimental data, the green pesticide composition for vegetable disease resistance and frost protection provided by this invention not only improves bactericidal toxicity and reduces the dosage of chemically synthesized agents, but also prevents frost damage to strawberries, promotes plant growth, enhances plant stress resistance, reduces disease severity, and lowers pesticide usage. This invention can also be used to control various diseases of corn and wheat, and promote their growth and enhance their stress resistance. This pesticide composition has significant application value in plant disease prevention and frost protection, a wide range of applications, and significant economic and social benefits. Detailed Implementation

[0015] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments.

[0016] Example 1: Preparation of a wettable powder (WP) with an active ingredient content of 50.5%.

[0017] The raw material ratio is as follows (parts by weight): Bacillus amyloliquefaciens fermentation broth 30, choline chloride 20, lentinan 0.5, wetting agent 10, and filler to make up to 100.

[0018] Preparation method: Weigh out Bacillus amyloliquefaciens, choline chloride, lentinan (master powder), wetting agent, and filler. First, mix the above raw materials evenly and dry them. Then, pulverize them by air jet milling to prepare a wettable powder with an effective ingredient content of 50.5%. The wettable powder can be granulated to prepare a water-dispersible granule. The wettable powder can be processed into a suspension by adding water and grinding with sand mill.

[0019] Example 2: Preparation of a wettable powder with an active ingredient content of 40.5%

[0020] The raw material ratio is as follows (parts by weight): Bacillus amyloliquefaciens fermentation broth 20, choline chloride 20, lentinan 0.5, wetting agent 10, and filler to make up to 100.

[0021] Preparation method: Weigh out Bacillus amyloliquefaciens, choline chloride, lentinan (master powder), wetting agent, and filler. First, mix the above raw materials evenly and dry them. Then, pulverize them by air jet milling to prepare a wettable powder with an effective ingredient content of 40.5%. The wettable powder can be granulated to prepare a water-dispersible granule. The wettable powder can be processed into a suspension by adding water and grinding with sand mill.

[0022] Example 3: Preparation of a wettable powder with an active ingredient content of 30.5%

[0023] The raw material ratio is as follows (parts by weight): Bacillus amyloliquefaciens fermentation broth 15, choline chloride 15, lentinan 0.5, wetting agent 10, and filler to make up to 100.

[0024] Preparation method: Weigh out Bacillus amyloliquefaciens, choline chloride, lentinan (master powder), wetting agent, and filler. First, mix the above raw materials evenly and dry them. Then, pulverize them by air jet milling to prepare a wettable powder with an effective ingredient content of 30.5%. The wettable powder can be granulated to prepare a water-dispersible granule. The wettable powder can be processed into a suspension by adding water and grinding with sand mill.

[0025] Example 4: Preparation of a wettable powder with an active ingredient content of 35.5%

[0026] The raw material ratio is as follows (parts by weight): Bacillus amyloliquefaciens fermentation broth 20, choline chloride 15, lentinan 0.5, wetting agent 10, and filler to make up to 100.

[0027] Preparation method: Weigh out Bacillus amyloliquefaciens, choline chloride, lentinan (master powder), wetting agent, and filler. First, mix the above raw materials evenly and dry them. Then, pulverize them by air jet milling to prepare a wettable powder with an effective ingredient content of 35.5%. The wettable powder can be granulated to prepare a water-dispersible granule. The wettable powder can be processed into a suspension by adding water and grinding with sand mill.

[0028] Example 5: Preparation of a wettable powder with an active ingredient content of 45.5%

[0029] The raw material ratio is as follows (parts by weight): Bacillus amyloliquefaciens fermentation broth 25, choline chloride 20, lentinan 0.5, wetting agent 10, and filler to make up to 100.

[0030] Preparation method: Weigh out Bacillus amyloliquefaciens, choline chloride, lentinan (master powder), wetting agent, and filler. First, mix the above raw materials evenly and dry them. Then, pulverize them by air jet milling to prepare a wettable powder with an effective ingredient content of 45.5%. The wettable powder can be granulated to prepare a water-dispersible granule. The wettable powder can be processed into a suspension by adding water and grinding with sand mill.

[0031] The filler in Examples 1-5 above is selected from at least one of silica, diatomaceous earth, attapulgite, clay, and earth clay. The wetting agent is selected from at least one of sodium dodecyl sulfate, sodium lignosulfonate, and lignosulfonate.

[0032] This invention relates to a series of wettable powders and formulations using Bacillus amyloliquefaciens, choline chloride, and lentinan as active ingredients. These formulations include: 30% Bacillus amyloliquefaciens wettable powder (WP), 30% choline chloride aqueous solution, 0.5% lentinan aqueous solution, 50.5% Bacillus amyloliquefaciens + choline chloride + lentinan (30:20:0.5) wettable powder, 40.5% Bacillus amyloliquefaciens + choline chloride + lentinan (20:20:0.5) wettable powder, and 30.5% Bacillus amyloliquefaciens + choline chloride + lentinan (15:15:0.5) wettable powder. All formulations were diluted 2000 times with water. A control was prepared by diluting 50.5% Bacillus amyloliquefaciens + choline chloride + lentinan 4000 times with water. These formulations were applied as a spray during vegetable transplanting and spring spraying to control anthracnose in vegetables. The spray volume was 30 kg per acre. The control effects on anthracnose are shown in Table 1.

[0033] Table 1. Field efficacy trial results of the Bacillus amyloliquefaciens + choline chloride + lentinan composition for controlling anthracnose in vegetables.

[0034]

[0035] *The drug concentrations in the table are for Bacillus amyloliquefaciens. The data were statistically analyzed using the new multiple range method. Data with the same letter are considered to have no significant difference.

[0036] Table 1 shows that field trials demonstrated that a 2000-fold dilution of the Bacillus amyloliquefaciens-choline chloride-lentinan wettable powder prepared according to this invention, applied as a spray at the time of tomato transplanting, achieved a control effect of 87.7-95.6% against tomato root rot and 87.4-94.4% against tomato diseases in spring, without adversely affecting tomato growth. At the same dosage of Bacillus amyloliquefaciens, the control efficacy of 50.5% WP (containing 30% tebuconazole) diluted 2000 times with water reached 96.5% and 94.0%, respectively, while the control efficacy of 30% Bacillus amyloliquefaciens WP diluted 2000 times with water reached 72.5% and 68.4%, respectively, significantly lower than the former. The control efficacy of 50.5% WP (containing 30% Bacillus amyloliquefaciens) diluted 4000 times with water reached 87.4% and 86.9% respectively, while the control efficacy of 30% Bacillus amyloliquefaciens WP diluted 2000 times with water reached 72.5% and 68.4% respectively, which were also significantly lower than the former.

[0037] This invention relates to the following formulations processed using Bacillus amyloliquefaciens, choline chloride, and lentinan as active ingredients: a 30% Bacillus amyloliquefaciens wettable powder (WP), a 30% choline chloride aqueous solution, a 0.5% lentinan aqueous solution, a 50.5% Bacillus amyloliquefaciens + choline chloride + lentinan (30:20:0.5) wettable powder, and a 40.5% Bacillus amyloliquefaciens + choline chloride + lentinan (20:20:0.5) wettable powder. The agent, 30.5% Bacillus amyloliquefaciens + choline chloride + lentinan (15:15:0.5) wettable powder, was diluted 2000 times with water. A control was set up with 50.5% Bacillus amyloliquefaciens + choline chloride + lentinan diluted 4000 times with water. The agent was sprayed on the Jinpeng tomato variety in the solar greenhouse of Shandong Agricultural University before the cold wave. The amount of pesticide solution sprayed per mu was 30 kg. The results were investigated after the cold wave. The results are shown in Table 2.

[0038] Table 2. Field trial results of the combination of Bacillus amyloliquefaciens, choline chloride, and lentinan for preventing tomato frost damage.

[0039]

[0040] *The drug concentrations in the table are for Bacillus amyloliquefaciens. The data were statistically analyzed using the new multiple range method. Data with the same letter are considered to have no significant difference.

[0041] Table 2 shows that the field trial demonstrated that the damage rate in the control group reached 42.5%. Compared with the control, the three compositions of 50.5% WP, 40.5% WP, and 30.5% WP prepared in this invention, diluted 2000 times with water and sprayed before the arrival of a cold wave, achieved a frost protection effect of 90.8%–95.8%, and the tomatoes grew normally. The frost protection effect of 50.5% WP (containing 30% tebuconazole) diluted 4000 times with water was also 90.1%. The frost protection effects of the four treatments of the three compositions were all greater than 90%, significantly higher than the frost protection effect of 30% choline chloride aqueous solution diluted 2000 times with water (80.7%), and also significantly higher than the frost protection effect of 0.5% lentinan aqueous solution diluted 2000 times with water (72.9%). In contrast, the frost protection effect of 30% Bacillus amyloliquefaciens WP diluted 2000 times with water was only 9.2%. This indicates that the frost protection effect of the tomato treatment prepared in this invention is very significant.

[0042] As can be seen from the above, the purpose of this invention is to provide a disease-resistant and frost-resistant pesticide composition for vegetables such as tomatoes. This composition can effectively prevent and control root rot, anthracnose, and powdery mildew in vegetables, and can also prevent frost damage, thereby improving the disease resistance and stress tolerance of vegetables. This composition not only enhances bactericidal toxicity but also prevents frost damage to vegetables, promotes plant growth, and strengthens plant stress resistance.

[0043] The method for using the above-mentioned pesticide composition to control diseases in strawberries and during the growth period of corn and wheat is as follows: At the initial stage of disease in strawberries, corn, and wheat, apply a wettable powder, water-dispersible granule, or suspension concentrate of Bacillus amyloliquefaciens • choline chloride • lentinan by mixing with water to form a suspension for spraying. This method has strong bactericidal and disease-preventing effects, with no adverse effects on crop growth, and significant economic and social benefits. Applying a wettable powder, water-dispersible granule, or suspension concentrate of Bacillus amyloliquefaciens • choline chloride • lentinan by mixing with water to form a suspension for spraying before a cold snap can effectively prevent frost damage to vegetables.

[0044] Those skilled in the art should understand that the above description is merely a specific embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A composition for applying Bacillus amyloliquefaciens BZ-32 and its metabolites to vegetable stress resistance, comprising an active ingredient and an adjuvant; wherein the active ingredient is composed of Bacillus amyloliquefaciens, choline chloride, and lentinan; wherein the weight percentages of the fermentation broth containing Bacillus amyloliquefaciens metabolites, choline chloride, and lentinan are 5-30% Bacillus amyloliquefaciens, 5-30% choline chloride, and 0.2-5.0% lentinan, respectively.

2. The composition of Bacillus amyloliquefaciens BZ-32 and its metabolites for stress resistance in vegetables as described in claim 1, characterized in that, The weight ratio of the fermentation broth containing Bacillus amyloliquefaciens metabolites, choline chloride, and lentinan is 10-30:10-30:0.2-2.

0.

3. The composition of Bacillus amyloliquefaciens BZ-32 and its metabolites for stress resistance in vegetables as described in claim 2, characterized in that, The weight ratio of the fermentation broth containing Bacillus amyloliquefaciens metabolites, choline chloride, and lentinan is 30:20:0.

5.

4. The composition of Bacillus amyloliquefaciens BZ-32 and its metabolites for stress resistance in vegetables as described in claim 1, characterized in that, The pesticide composition is a suspension concentrate, wettable powder, or water-dispersible granule.

5. Use of the pesticide composition according to any one of claims 1-4 in the prevention and control of seedling diseases in vegetables and / or in the resistance of vegetables to stress.

6. The use as described in claim 5, characterized in that, The diseases affecting vegetable seedlings include root rot, anthracnose, and powdery mildew.

7. The use as described in claim 5, characterized in that, The application of the vegetable's stress resistance is for adverse stress caused by excessively low winter temperatures or late spring frosts.

Images