CN108485997A - A kind of method that biological control cuts down Wolfberry Aphid and improves production performance - Google Patents

Abstract

The invention discloses a kind of methods biological control abatement Wolfberry Aphid and improve production performance, including：The hereinafter referred to as Sb of bacillus subtilis CICC 10339, Sb is cultivated, and Sb bacterium solutions are made；Corn flour uncooked amylum enzyme is digested, micropore corn flour is prepared into；Sb is embedded into the micropore of micropore corn flour, micropore corn flour Sb bacterium solutions are made；Sb microbial inoculums are made in micropore corn flour Sb bacterium solutions；Using Sb bacterium solutions as leavening, Sb bio-fertilizers are made；Sb microbial inoculums are sprayed on matrimony vine plant surface, effectively cut down Wolfberry Aphid；Sb bio-fertilizers are imposed on into matrimony vine plant root, effectively cut down Wolfberry Aphid.The present invention Sb Developing Green biological controls of production chitinase, and use a variety of comprehensive means of supplementing out economy, Sb microbial inoculums and Sb bio-fertilizers and use is then synergistic, to effectively abatement Wolfberry Aphid and improve matrimony vine production performance.

Description

A method for biological control to reduce wolfberry aphids and improve production performance

technical field

The invention relates to the field of biological control, in particular to a method for reducing wolfberry aphids and improving production performance through biological control.

Background technique

Lycium barbarum has pests and diseases in various stages of growth, and aphids are one of the main pests. When the air humidity is higher than 75%, it is not conducive to the reproduction of Lycium barbarum aphids. Therefore, Lycium barbarum aphids are more harmful in dry seasons. Excessive application of nitrogen fertilizers in production will also lead to the occurrence of aphid pests. Lycium barbarum aphids live through the winter with their eggs on the branches or at the roots of wolfberry plants on the ground. They start to damage wolfberry after they germinate in April, and flourish in May. Flowering and fruiting and growth and development cause the output of Lycium barbarum fruit to reduce by 30%-70%, the quality of the product is greatly reduced, and the commodity value is seriously affected. In the hot summer, the population of insects decreased, and then increased again in autumn, and the second peak appeared in September.

Although pesticides can control the damage of aphids, long-term use will cause aphids to develop drug resistance, resulting in increased dosage of pesticides and increased pesticide residues in wolfberry products, thus affecting the quality of wolfberries.

In addition to the application of pesticides, various methods can be used to reduce the harm of wolfberry aphids, including: deep plowing before the garden is built, thorough cleaning of the wolfberry garden, reasonable intercropping after the garden is built, reasonable irrigation and fertilization (do not use too much nitrogen fertilizer), and timely pruning Severely damaged branches; use physical means, such as trapping with sweet and sour liquid, trapping with yellow boards or insecticidal lamps, hanging silver-gray plastic strips to drive away, pouring water on the crown of the tree to wash away aphids; spraying ammonia fertilizer or penetrating agent to kill aphids at high temperatures ; Protection of natural enemies of aphids; application of botanical pesticides pyrethrum, matrine, anisein, toosendanin, garlic, etc.; application of microbial source insecticides, fungicides and herbicides; application of mineral source pesticides such as crystal lime sulfur and stone Sulfur compound (an inorganic preparation that can not only kill bacteria, but also kill insects and mites), sulfur suspending agent.

Biological control is the first choice to effectively control pests and ensure the quality of wolfberry products, and microbial control is the most effective and feasible option. Aphids belong to the insects of the order Homoptera, and chitin accounts for 25-40% of the dry weight of the body wall, which constitutes the main structure of the body wall and the peritrophic cavity membrane, and supports and protects the body. Therefore, microorganisms with high chitinase production can destroy the body structure of aphids by degrading chitin, becoming an effective biological control method.

Bacillus subtilis is recognized as a food-grade safe microorganism (generally regarded as safety, GRAS), which can be widely colonized in natural environments such as soil, plant surface and interior, thus becoming an ideal biological control microorganism. The present invention adopts Bacillus subtilis (CICC 10339) (hereinafter referred to as Sb) of high-yield chitinase to carry out biological control of wolfberry aphids. At the same time, because Sb degrades complex substrates, it also plays a role in understanding phosphorus and potassium. The combined use of Sb inoculum and Sb biological fertilizer can effectively reduce the damage of wolfberry aphids and improve the production performance of wolfberry.

Contents of the invention

The invention provides a method for biological control and reduction of wolfberry aphids and improvement of wolfberry production performance, which is efficient, simple and environment-friendly.

In order to achieve the above object, the present invention adopts the following technical solutions: by preparing Sb bacterial liquid and Sb bacterial agent of high-yield chitinase, the Sb bacterial liquid is used to prepare biological fertilizer, the Sb bacterial agent sprays Lycium barbarum plants, and in the Sb bacterial liquid , Sb inoculum and bio-fertilizer preparation process, give full play to the synergistic effect of auxiliary materials and the growth-promoting effect of Sb, thereby effectively reducing wolfberry aphids and improving the production performance of wolfberry.

A method for biological control and reduction of wolfberry aphids and improvement of wolfberry production performance, comprising the following steps:

1) Bacillus subtilis CICC 10339 is hereinafter referred to as Sb, and the Sb is cultivated to make a Sb bacterial solution with a density of 10 ⁹ to 10 ¹⁰ cfu/mL;

2) enzymolyzing the corn flour with raw amylase to prepare microporous corn flour;

3) adding the microporous corn flour prepared in step 2) to the Sb bacterial liquid prepared in step 1), embedding with vibration to make the microporous corn flour Sb bacterial liquid;

4) the microporous corn flour Sb bacterial liquid prepared in step 3) is made into Sb microbial agent;

5) using the Sb bacterium liquid prepared in step 1) as a starter to make Sb biological fertilizer;

6) Spray the Sb bacterial agent prepared in step 4) on the surface of the Lycium barbarum plant, and at the same time, apply the Sb biological fertilizer prepared in the step 5) to the root of the Lycium barbarum plant to effectively reduce the Lycium barbarum aphid.

Following as preferred technical scheme of the present invention:

In step 1), the Sb is cultured, including: the starting density of Sb is 10 ⁶ to 10 ⁷ /mL, inoculated into the liquid medium, the inoculum amount is 2% to 4%, and the culture is shaken at 36°C to 38°C for 22h to After 26 hours, reach the Sb bacterial liquid with a bacterial liquid density of 10 ⁹ -10 ¹⁰ cfu/mL. The inoculum size refers to the ratio of the volume of the seed solution transferred to the volume of the culture solution after inoculation, that is, the ratio of the volume of Sb transferred to the volume of the liquid medium.

More preferably, the Sb is cultured, including: the starting density of Sb is 10 ⁶ to 10 ⁷ /mL, inoculated into the liquid culture medium with an inoculum size of 3%, and cultured with shaking at 37°C for 24 hours until the density of the bacterial liquid is 10 9 to 10 ⁹ . 10 ¹⁰ cfu/mL of Sb bacteria solution.

When Sb is cultured, the liquid culture medium, in 1L, includes the following components:

Adjust the pH to 6.5-7.5, and sterilize at 121°C for 20 minutes.

Further preferably, when Sb is cultured, the liquid culture medium, in 1L, includes the following components:

Adjust the pH to 7.0, and sterilize at 121°C for 20 minutes.

In step 1), Bacillus subtilis CICC 10339 is a commercially available product, which can be purchased from the China Industrial Microbial Culture Collection Center (CICC), address: Building 6, Yard 24, Jiuxianqiao Middle Road, Chaoyang District, Beijing; strain maintenance number for CICC 10339.

In step 2), corn flour is enzymatically hydrolyzed with raw amylase to prepare microporous corn flour, which includes: adding water to corn flour and stirring to make corn milk, adding alpha-amylase (Ec.3.2.1.1) and glucoamylase ( EC.3.2.1.3) composed of raw amylase, cultured by shaking, the starch in the corn flour granules is partially hydrolyzed to form holes, and after centrifugal drying, it becomes microporous corn flour.

The quality of the corn flour and water is 50:40-60. Further preferably, the concentration of corn milk is 50%, that is, the mass of corn flour and water is 50:50.

The enzyme dosage of α-amylase added to corn milk is 5U/L, and the enzyme dosage of glucoamylase added to corn milk is 500U/L. The α-amylase and glucoamylase adopt the existing technology and meet the national food safety standard - food additives - enzymes for food industry (GB 1886.174-2016). Enzyme activity unit U: 1 U is the amount of enzyme that hydrolyzes the substrate in 1 minute to release 1 μmol of product (calculated as reducing sugar) under the conditions of 37°C and pH 6.5.

The shaking culture conditions are as follows: shaking culture at 35° C. to 39° C. for 20 hours to 28 hours, and shaking culture at 37° C. for 24 hours.

In step 3), the weight ratio of the volume of the Sb bacterial liquid to the microporous corn flour is 100L: 3-10Kg. Further preferably, the weight ratio of the volume of the Sb bacterial liquid to the microporous corn flour is 100L:5Kg.

The conditions for the embedding by shaking are: 18° C. to 22° C. for 0.5 h to 2 h, more preferably, 20° C. for 1 hr.

In step 4), glutinous rice flour is used as the wall material, jasmine leaf powder and hyaluronic acid are added, and the microporous corn flour Sb bacterial liquid is spray-dried to make the Sb microbial agent;

Adopt the following components by weight:

The conditions of the spray drying are: the inlet temperature is 160°C-180°C, and the outlet temperature is 70°C-90°C.

More preferably, glutinous rice flour is used as the wall material, jasmine leaf powder and hyaluronic acid are added, and the microporous corn flour Sb bacterial liquid is spray-dried to make the Sb bacterial agent. The formula and process are as follows: make the relevant components into a slurry in proportion After feeding, make Sb bacterial agent.

The relevant components are as follows:

The conditions of the spray drying are: inlet temperature 170°C, outlet temperature 80°C.

In step 5), the preparation of Sb biological fertilizer adopts a composting process, including: stirring each component of the compost evenly, and spraying the Sb bacterial liquid prepared in step 1), the spraying amount is 1%, and the spraying amount refers to the Sb bacterial liquid Volume L/ compost weight Kg, shelter from rain and direct sunlight, and compost for 6-10 days to get Sb bio-fertilizer.

Each component of compost is proportioned according to the following parts by weight:

Further preferably, each component of compost is proportioned according to the following parts by weight:

In step 6), the Sb bacterial agent is sprayed on the wolfberry plant surface, including:

After the Sb bacterial agent was diluted with water at a weight ratio of 1:100-200, it was sprayed on the surface of Lycium barbarum plants at a rate of 1000 mL per Lycium barbarum plant, sprayed once a week for 4 consecutive weeks.

The Sb bio-fertilizer is applied to each Lycium barbarum plant in an amount of 1-3 kg.

Compared with prior art, the present invention has following advantage:

Sb produces chitinase, which strongly destroys the insect body wall and peritrophic membrane, and has a significant killing effect on insects. It is a green biological control strategy.

In the present invention, Sb is made into a bacterial liquid, then made into a bacterial agent and fermented into a biological fertilizer, the bacterial agent is sprayed on the Lycium barbarum plant, and the biological fertilizer is applied to the root of the Lycium barbarum plant, both of which can effectively reduce the Lycium barbarum aphid, and play a role in plant promotion. It can be used to improve the production performance of wolfberry.

In the process of making Sb into bacterial liquid and preparing Sb bio-fertilizer, shrimp powder rich in chitin or chitin was included in the fermentation substrate, which induced Sb to produce high chitinase.

When the Sb bacterial liquid is made into the Sb bacterial agent by spray drying, the Sb is embedded in the porous cornmeal first, which helps to improve its survival rate during spray drying. Spray drying uses glutinous rice flour as the wall material, adding jasmine leaf powder and hyaluronic acid. Glutinous rice flour increases the adhesion of the bacterial agent when spraying. Hyaluronic acid inhibits insect digestive enzymes. Jasmine leaves are rich in derivatives such as jasmonic acid and its methyl ester, which induce protease inhibitor protein synthesis, thereby inhibiting insect digestion.

When fermenting Sb compost into bio-fertilizer, the compost formula is: straw, soybean meal, bran, plant ash, and shrimp meal. Plant ash contributes to high-density fermentation of Sb. Soybean meal not only provides nitrogen, but also contains trypsin inhibitors, which can inhibit insect digestion. Shrimp powder is rich in chitin, namely chitin, which induces high production of chitinase by Sb. Shrimp powder is also rich in fertilizers such as phosphorus, potassium, and nitrogen. Sb in biofertilizer also has the functions of dissolving phosphorus, dissolving potassium, and biocontrol.

In summary, the present invention uses chitinase-producing Sb to carry out green biological control, and adopts a variety of comprehensive supplementary means, such as the fermentation substrate component shrimp powder rich in chitin to induce Sb to produce high chitin Enzymes and plant ash are helpful for high-density fermentation of Sb, embedding Sb in porous cornmeal to improve its survival rate during spray drying, components used to prepare Sb inoculants have auxiliary effects for reducing pests, Sb The bio-fertilizer components have the effect of promoting plant growth, and the combination of Sb inoculum and Sb bio-fertilizer will have synergistic effects, thereby effectively reducing wolfberry aphids and improving the production performance of wolfberry.

Detailed ways

Example 1

1) High-density culture of chitinase-producing Sb (Bacillus subtilis CICC 10339), the starting density of Sb is 2.13× ¹⁰⁶ /mL, inoculated into liquid medium (1 L, glucose 11g, soybean powder 20g , shrimp powder 1g, the balance is water, adjust pH = 6.5, sterilize at 121°C for 20min), inoculum amount is 2%, shake culture at 36°C for 22h, the density of bacteria liquid is 3.85×10 ⁹ cfu/mL, and high density Sb bacteria solution.

2) Add corn flour to water and stir to make corn milk. The concentration of corn milk is 50% (that is, the mass ratio of corn flour to water is 50:50). Add α-amylase (EC.3.2.1.1) and glucoamylase (EC. 3.2.1.3) composed of raw amylase (enzyme dosages are 5U/L and 500U/L respectively), cultured with shaking at 37°C for 24hrs, the starch in the corn flour granules was partially hydrolyzed to form holes, dried at 120°C after centrifugation, and became micro Hole cornmeal.

3) Add microporous corn flour to the Sb bacterial solution to a concentration of 5% (W/V) (that is, the weight ratio of the volume of the Sb bacterial solution to the microporous corn flour is 100L: 5Kg), shake and embed at 20°C for 1 hr , become microporous corn flour Sb bacteria liquid.

4) Use glutinous rice flour as the wall material, add jasmine leaf powder and hyaluronic acid, and spray-dry the microporous cornflour Sb bacterial solution to prepare the Sb bacterial agent. After the relevant components (10 parts of microporous corn flour Sb bacterial liquid, 1 part of glutinous rice flour, 0.1 part of jasmine leaf powder, 0.1 part of hyaluronic acid) were made into slurry, spray drying (inlet temperature 170 ℃, outlet temperature 80 ℃ ) to make Sb bacterial agent.

5) Preparation of Sb biofertilizer by composting: Stir the components of the compost (50 parts of straw, 20 parts of soybean meal, 18 parts of bran, 10 parts of plant ash, and 2 parts of shrimp powder), and spray the prepared Sb bacterial liquid, spray The amount is 1% (Sb bacterial liquid volume L/compost weight Kg), sheltered from rain and sunlight, and composted for 6 days to obtain Sb bio-fertilizer.

6) The Sb bacterial agent was diluted with water at a weight ratio of 1:100, and then sprayed on the surface of Lycium barbarum plants. The spraying amount (based on the weight of the bacterial agent) was 1000mL per Lycium barbarum plant, and sprayed once a week for 4 consecutive weeks.

7) Sb biological fertilizer was applied to each Lycium barbarum plant in an amount of 1kg.

Example 2

1) High-density culture of chitinase-producing Sb, the starting density of Sb is 1.77× ¹⁰⁶ /mL, inoculated into liquid medium (1 L, glucose 17g, soybean powder 30g, shrimp powder 3g, the balance water, adjusted to pH = 7.5, sterilized at 121°C for 20 minutes), inoculum size was 4%, cultured with shaking at 38°C for 26 hours, the density of the bacterial liquid was 4.12×10 ⁹ cfu/mL, and a high-density Sb bacterial liquid was obtained.

2) Add corn flour to water and stir to make corn milk. The concentration of corn milk is 50% (that is, the mass ratio of corn flour to water is 50:50). Add α-amylase (EC.3.2.1.1) and glucoamylase (EC. 3.2.1.3) composed of raw amylase (enzyme dosages are 5U/L and 500U/L respectively), cultured with shaking at 37°C for 24hrs, the starch in the corn flour granules was partially hydrolyzed to form holes, dried at 120°C after centrifugation, and became micro Hole cornmeal.

3) Add microporous corn flour to the concentration of 5% (W/V) in the Sb bacterial solution, shake and embed at 20° C. for 1 hr to become the microporous corn flour Sb bacterial solution.

4) Use glutinous rice flour as the wall material, add jasmine leaf powder and hyaluronic acid, and spray-dry the microporous cornflour Sb bacterial solution to prepare the Sb bacterial agent. After the relevant components (10 parts of microporous corn flour Sb bacterial liquid, 1 part of glutinous rice flour, 0.1 part of jasmine leaf powder, 0.1 part of hyaluronic acid) were made into slurry, spray drying (inlet temperature 170 ℃, outlet temperature 80 ℃ ) to make Sb bacterial agent.

5) Preparation of Sb biofertilizer by composting: Stir the components of the compost (50 parts of straw, 20 parts of soybean meal, 18 parts of bran, 10 parts of plant ash, and 2 parts of shrimp powder), and spray the prepared Sb bacterial liquid, spray The amount is 1% (Sb bacterial liquid volume/compost weight), sheltered from rain and direct sunlight, and composted for 10 days to obtain Sb bio-fertilizer.

6) The Sb bacterial agent was diluted with water at a weight ratio of 1:200, and then sprayed on the surface of Lycium barbarum plants. The spraying amount (based on the weight of the bacterial agent) was 1000mL per Lycium barbarum plant, and sprayed once a week for 4 consecutive weeks.

7) Sb biological fertilizer was applied to each Lycium barbarum plant in an amount of 3kg.

Example 3

1) High-density culture of chitinase-producing Sb, the starting density of Sb is 1.95× ¹⁰⁶ /mL, inoculated into liquid medium (1 L, glucose 14g, soybean powder 25g, shrimp powder 2g, the balance water, adjusted to pH = 7.0, sterilized at 121°C for 20 minutes), with an inoculum size of 3%, shake cultured at 37°C for 24 hours, and the density of the bacterial liquid was 6.24×10 ⁹ cfu/mL to obtain a high-density Sb bacterial liquid.

2) Add corn flour to water and stir to make corn milk. The concentration of corn milk is 50% (that is, the mass ratio of corn flour to water is 50:50). Add α-amylase (EC.3.2.1.1) and glucoamylase (EC. 3.2.1.3) composed of raw amylase (enzyme dosages are 5U/L and 500U/L respectively), cultured with shaking at 37°C for 24hrs, the starch in the corn flour granules was partially hydrolyzed to form holes, dried at 120°C after centrifugation, and became micro Hole cornmeal.

3) Add microporous corn flour to the concentration of 5% (W/V) in the Sb bacterial solution, shake and embed at 20° C. for 1 hr to become the microporous corn flour Sb bacterial solution.

4) Use glutinous rice flour as the wall material, add jasmine leaf powder and hyaluronic acid, and spray-dry the microporous cornflour Sb bacterial solution to prepare the Sb bacterial agent. After the relevant components (10 parts of microporous corn flour Sb bacterial liquid, 1 part of glutinous rice flour, 0.1 part of jasmine leaf powder, 0.1 part of hyaluronic acid) were made into slurry, spray drying (inlet temperature 170 ℃, outlet temperature 80 ℃ ) to make Sb bacterial agent.

5) Preparation of Sb biofertilizer by composting: Stir the components of the compost (50 parts of straw, 20 parts of soybean meal, 18 parts of bran, 10 parts of plant ash, and 2 parts of shrimp powder), and spray the prepared Sb bacterial liquid, spray The amount is 1% (Sb bacterial liquid volume/compost weight), sheltered from rain and sunlight, and composted for 8 days to obtain Sb bio-fertilizer.

6) The Sb bacterial agent was diluted with water at a weight ratio of 1:150, and then sprayed on the surface of Lycium barbarum plants. The spraying amount (based on the weight of the bacterial agent) was 1000mL per Lycium barbarum plant, and sprayed once a week for 4 consecutive weeks.

7) Sb biological fertilizer was applied to each Lycium barbarum plant in an amount of 2kg.

Comparative example 1

1) Corn flour is mixed with water to make corn milk, the concentration of corn milk is 50%, and raw amylase composed of α-amylase (EC.3.2.1.1) and glucoamylase (EC.3.2.1.3) is added (the enzyme dosage is respectively 5U/L and 500U/L), cultured with shaking at 37°C for 24hrs, the starch in the cornflour granules was partially hydrolyzed to form holes, and dried at 120°C after centrifugation to become microporous cornflour.

2) Make the relevant components (10 parts of microporous corn flour, 1 part of glutinous rice flour, 0.1 part of jasmine leaf powder, 0.1 part of hyaluronic acid, 20 parts of water) into a slurry, spray dry (inlet temperature 170 ℃, outlet temperature 80° C.) to make contrast agent-I.

3) The composting process is adopted for the preparation of non-Sb biological fertilizer: Stir the components of the compost (50 parts of straw, 20 parts of soybean meal, 18 parts of bran, 10 parts of plant ash, and 2 parts of shrimp powder), avoid rain and direct sunlight, and compost 8d, obtain non-Sb biological fertilizer.

4) After the contrast agent-I was diluted with water according to the weight ratio of 1:150, it was sprayed on the surface of Lycium barbarum plants, and the spraying amount (calculated by the weight of bacterial agent) was 1000mL per Lycium barbarum plant, and sprayed once a week for 4 consecutive weeks.

5) Non-Sb biological fertilizer was applied to each Lycium barbarum plant in an amount of 2kg.

Comparative example 2

1) Corn flour is mixed with water to make corn milk, the concentration of corn milk is 50%, and raw amylase composed of α-amylase (EC.3.2.1.1) and glucoamylase (EC.3.2.1.3) is added (the enzyme dosage is respectively 5U/L and 500U/L), cultured with shaking at 37°C for 24hrs, the starch in the cornflour granules was partially hydrolyzed to form holes, and dried at 120°C after centrifugation to become microporous cornflour.

2) The related components (10 parts of microporous corn flour, 20 parts of water) were made into a slurry, and then spray-dried (inlet temperature 170°C, outlet temperature 80°C) to prepare contrast agent-II.

3) The composting process is adopted for the preparation of non-Sb biological fertilizer: Stir the components of the compost (50 parts of straw, 20 parts of soybean meal, 18 parts of bran, 10 parts of plant ash, and 2 parts of shrimp powder), avoid rain and direct sunlight, and compost 8d, obtain non-Sb biological fertilizer.

4) The control agent-II was diluted with water at a weight ratio of 1:150, and then sprayed on the surface of Lycium barbarum plants. The spraying amount (calculated by the weight of bacterial agent) was 1000mL per Lycium barbarum plant, and sprayed once a week for 4 consecutive weeks.

5) Non-Sb biological fertilizer was applied to each Lycium barbarum plant in an amount of 2kg.

Embodiment and comparative example result contrast

As shown in the examples and Table 1, there is high activity of chitinase in the bacteria liquid of the example, high density of Sb in the bacterial agent, and high activity of chitinase and high density of Sb in the prepared Sb bio-fertilizer, However, the bacterial agent of the control example and the prepared bio-fertilizer basically had no Sb and chitinase activity, showing a close correlation between high density Sb and high chitinase activity. The wolfberry aphid situation in the embodiment is significantly lower than that of the control example, while the yield per plant of the wolfberry production performance is significantly higher than that of the control example, indicating that the Sb-rich bacterial agent and biological fertilizer have the effect of reducing the wolfberry aphid and improving the production performance of the wolfberry.

The aphid situation of Lycium barbarum in Control Example 1 was significantly lower than that of Control Example 2, and the yield per plant of Lycium barbarum production performance was significantly higher than that of Control Example 2, indicating that although there is no Sb in the bacterial agent and biological fertilizer, the hyaluronic acid and jasmine in the bacterial agent The jasmonic acid contained in the leaf powder still has certain effects on reducing the aphids of Lycium barbarum and improving the production performance of Lycium barbarum. The former inhibits insect digestive enzymes, and the latter induces protein synthesis of protease inhibitors, thereby inhibiting the digestive ability of insects.

Determination of chitinase activity: refer to "Wu Zhigang. Isolation of chitin/chitosanase-producing bacteria, research on fermentation conditions and enzymatic properties [D]. Hangzhou: Zhejiang University, 2003." Take 5mL of Sb bacteria liquid, centrifuge at 5000rpm for 10min; take 0.5mL of supernatant, add 1mL of McIllvaine buffer, 0.5mL of 3% colloidal chitin, bathe in water at 37℃ for 30min, then take a boiling water bath for 10min, take 1mL, centrifuge at 5000rpm for 5min, add Stop the reaction with 0.5mL DNS reagent; cool immediately after boiling water bath for 5min, add 4mL deionized water, measure OD _540nm value; take N-acetylglucose content as the abscissa, OD _540nm value as the ordinate, draw the standard curve, blank tube as zero. Under the conditions of 37°C and pH 7.0, the amount of enzyme needed to produce reducing sugar equivalent to 1 μmol N-acetylglucosamine per minute is defined as 1 enzyme activity unit (U).

Determination of Sb density: according to "GB/T 26428-2010 Detection of Bacillus subtilis in microbial preparations for feeding".

Evaluation of Lycium barbarum aphids situation: In Lycium barbarum grove, 8 plots (4 Lycium barbarum plants per plot) are set up in each embodiment or control example, and 8 plots of blank control are assumed, totally 48 plots. Five branches of each Lycium barbarum plant were randomly surveyed in 5 orientations: east, south, west, north and middle, and the amount of aphids per branch was recorded before and 3 days after the test. Infestation was assessed according to the following formula:

Control effect = (1-CK ₀ ×PT ₁ /CK ₁ ×PT ₀ )×100%

CK ₀ : the number of live worms before the test in the blank control area.

CK ₁ : The number of live worms after the test in the blank control area.

PT ₀ : the number of living insects before the test of the embodiment or the control example.

PT ₁ : the number of living insects after the test of the embodiment or the control example.

Determination of yield per plant of Lycium barbarum: In the Lycium barbarum grove, 8 plots (4 Lycium barbarum plants per plot) were set up in each embodiment or control example, and each plot was weighed after the fresh fruit was picked.

The detection sample number of the Lycium barbarum aphid situation and the yield per plant of each embodiment or comparative example is 8.

The comparison of the bacterial liquid, microbial agent, biological fertilizer and action effect prepared by the embodiment and the comparative example is shown in Table 1.

Bacterial liquid, bacterium agent, bio-fertilizer and action effect contrast prepared by the embodiment of table 1 and comparative example

Claims (10)

1. a kind of method that biological control cuts down Wolfberry Aphid and improves matrimony vine production performance, which is characterized in that including following step Suddenly：

1) the hereinafter referred to as Sb of bacillus subtilis CICC 10339, Sb is cultivated, and density 10 is made⁹~10¹⁰Cfu/mL's Sb bacterium solutions；

2) corn flour uncooked amylum enzyme is digested, is prepared into micropore corn flour；

3) the micropore corn flour that in the Sb bacterium solutions for preparing step 1) prepared by addition step 2), oscillation embedding, is made micropore corn Powder Sb bacterium solutions；

4) Sb microbial inoculums are made in micropore corn flour Sb bacterium solutions prepared by step 3)；

5) Sb bio-fertilizers are made as leavening in the Sb bacterium solutions prepared using step 1)；

6) Sb microbial inoculums prepared by step 4) are sprayed on matrimony vine plant surface, meanwhile, Sb bio-fertilizers prepared by step 5) are imposed on Matrimony vine plant root.

2. the method that biological control according to claim 1 cuts down Wolfberry Aphid and improves matrimony vine production performance, feature It is, in step 1), Sb is cultivated, including：Sb set out density be 10⁶~10⁷A/mL, is seeded in fluid nutrient medium, Inoculum concentration is 2%~4%, 36 DEG C~38 DEG C shaken cultivation 22h~26h, until bacterium solution density is 10⁹~10¹⁰The Sb bacterium of cfu/mL Liquid.

3. the method that biological control according to claim 2 cuts down Wolfberry Aphid and improves matrimony vine production performance, feature It is, when Sb is cultivated, the fluid nutrient medium, in terms of 1L, including following components：

Adjust pH=6.5~7.5,121 DEG C of sterilizing 20min.

4. the method that biological control according to claim 1 cuts down Wolfberry Aphid and improves matrimony vine production performance, feature It is, in step 2), corn flour uncooked amylum enzyme is digested, micropore corn flour is prepared into, including：Corn flour adds water stirring to be tuned into Corn milk, is added the uncooked amylum enzyme that is made of alpha-amylase and carbohydrase, shaken cultivation, and the starch in corn powder particles is by part Hole is hydrolyzed to form, becomes micropore corn flour after centrifugation drying.

5. the method that biological control according to claim 1 cuts down Wolfberry Aphid and improves matrimony vine production performance, feature It is, in step 3), the volume of the Sb bacterium solutions is 100L with the weight ratio of micropore corn flour：3Kg~10Kg.

6. the method that biological control according to claim 1 cuts down Wolfberry Aphid and improves matrimony vine production performance, feature It is, in step 3), the condition of the oscillation embedding is：18 DEG C~22 DEG C oscillation embedding 0.5h~2h.

7. the method that biological control according to claim 1 cuts down Wolfberry Aphid and improves matrimony vine production performance, feature It is, in step 4), using glutinous rice flour as wall material, adds arabian jasmine leaf powder and hyaluronic acid, micropore corn flour Sb bacterium solutions are passed through Sb microbial inoculums are made in spray drying；

Using the component of following parts by weight：

The condition of the spray drying is：160 DEG C~180 DEG C of inlet temperature, 70 DEG C~90 DEG C of outlet temperature.

8. the method that biological control according to claim 1 cuts down Wolfberry Aphid and improves matrimony vine production performance, feature It is, in step 5), prepared by Sb bio-fertilizers uses composting process, including：Compost each component is stirred evenly, step 1) institute is sprayed The Sb bacterium solutions of preparation, the amount of spraying are 1%, and the amount of spraying refers to Sb bacterium solution volume L/ compost weight Kg, direct sunlight of taking shelter from rain, compost 6d~10d is to get to Sb bio-fertilizers；

Compost each component presses following weight：

9. the method that biological control according to claim 1 cuts down Wolfberry Aphid and improves matrimony vine production performance, feature It is, in step 6), Sb microbial inoculums are sprayed on matrimony vine plant surface, including：Sb microbial inoculums are according to weight ratio 1:100~200 is dilute with water After releasing, matrimony vine plant surface is sprayed, the amount of spraying is every plant of matrimony vine 1000mL, is sprayed weekly once, continuous 4 weeks.

10. the method that biological control according to claim 1 cuts down Wolfberry Aphid and improves matrimony vine production performance, feature It is, in step 6), the Sb bio-fertilizers impose on every plant of matrimony vine according to the amount of 1~3kg.