CN101805700B

CN101805700B - Method for producing white muscardine fungi by using silkworm pupae and application thereof

Info

Publication number: CN101805700B
Application number: CN 201010132214
Authority: CN
Inventors: 吴洪生
Original assignee: 吴洪生
Current assignee: GUANGXI YANGGUI BIOTECHNOLOGY CO., LTD.
Priority date: 2010-03-25
Filing date: 2010-03-25
Publication date: 2012-12-26
Anticipated expiration: 2030-03-25
Also published as: CN101805700A

Abstract

The invention relates to a method for producing white muscardine fungi by using silkworm pupae, which comprises the following steps of: firstly, preparing a culture medium which contains the silkworm pupae or a mixture of which weight ratio of the silkworm pupae to water is 20-50:80-50, or preparing a cocoon cooking liquid or a mixture of which the weight ratio of the silkworm pupae to the cocoon cooking liquid is 10-40:90-60; secondly, sterilizing the culture medium and then putting the culture medium into a culture vessel; thirdly, cooling the culture medium to 30 to 32 DEG C; fourthly, inoculating the white muscardine fungi, wherein the inoculated white muscardine fungi accounts for 2 to 30 percent of the weight of the culture medium; fifthly, passing sterile air into the mixture, keeping the culture temperature between 20 and 30 DEG C; sixthly, after 5 to 7 days of culture, adding wheat bran which accounts for 5 to 20 percent of the weight of the total cultures, adding clay or attapulgite which accounts for 10 to 30 percent of the weight of the total cultures into the mixture, and stirring the mixture uniformly; seventhly, performing plate and frame type filter pressing on the mixture or drying the mixture at the temperature of 35 DEG C; and finally, crushing the mixture to prepare a solid powder product. The method fully uses biological resources, changes wastes into valuable, increases the incomings of silkworm raisers, and simultaneously can realize indoor industrialization and large-scale production very easily by using a modern microbial technical skill, and a prepared white muscardine fungi preparation can be used for preventing and controlling agricultural pests.

Description

Utilize silkworm chrysalis to produce method and the application thereof of muscardine

One, technical field

The invention belongs to agricultural technology field, relate in particular to the working method and the application of one type of agricultural microorganism sterilant.

Two, background technology

China is world silkworm sparetime university state, annual more than 1,000 ten thousand tons of the silk cocoons of producing, and annual filature factory produces a large amount of sub product silkworm chrysalises and cocoon cooking liquid in process of cocoon, pass into disuse serious environment pollution and stench basically.Contain nutritive substances such as amounts of protein in silkworm chrysalis and the cocoon cooking liquid, utilize silk cocoon or cocoon cooking liquid to produce muscardine and products thereof, report is not arranged at present as yet, can silk cocoon and cocoon cooking liquid be made full use of, reduce environmental pollution.The muscardine preparation of producing simultaneously is the friendly type biological pesticide of a kind of fine environment.Be agricultural herd and gardening on good biological pesticide, be used to prevent and treat the insect of farm crop and vegetables, flowers and trees, Chinese medicinal materials, effect is fine; Both single-minded killing off the insect pests; Do not injure natural enemy again, more can not produce harm and contaminate environment, help the human kind sustainable development and the eubiosis people and animals.

Three, summary of the invention

Goal of the invention: the object of the invention is to provide a kind of method and the application of muscardine in control of agricultural pest that utilizes waste silkworm chrysalis, cocoon cooking liquid and the pupa slag production muscardine of silkworm industry and silk mill, through the stiff bacteria preparation of industrial production of white.

Technical scheme: utilize silkworm chrysalis to produce the method for muscardine, first preparing culture medium, said substratum contains silkworm chrysalis; Or silkworm chrysalis and water is by weight the mixture of 20-50: 80-50, or boils silk cocoon liquid, or silkworm chrysalis and boil the mixture of silk cocoon liquid by weight 10-40: 90-60; With inserting behind the medium sterilization in the culture vessel, substratum is cooled to 30～32 ℃ again, the inoculation muscardine; The weight ratio that the muscardine of inoculation accounts for substratum is 2%wt～30%wt, and passes to sterile air, and keeping culture temperature is 20～30 ℃; Cultivate after 5～7 days, add the 5-20% wheat bran of total culture weight, clay or the attapulgite of 10-30%, mix thoroughly; Filter press or 35 ℃ of oven dry are pulverized and are processed the solid powdery product.

Above-mentioned sterilising conditions is 115～130 ℃, 0.1～0.15MPa, and the time is 30～90 minutes.

Before the above-mentioned inoculation substratum is cooled to 30 ℃.

The weight ratio that the muscardine of above-mentioned inoculation accounts for substratum is 10%.

After the above-mentioned inoculation, keeping culture temperature is 30 ℃.

Beneficial effect: the present invention compares prior art and has following beneficial effect: the present invention utilizes silkworm chrysalis, boils silk cocoon liquid and pupa slag production muscardine; Can satisfy the needs in the production, can make full use of in the traditional habit silkworm chrysalis again, boil the Biological resources that silk cocoon liquid and pupa slag are thrown away when rubbish, turn waste into wealth; Increase silkworm raiser's income; Utilize modern microbiological processes technology can be easy to realize indoor industrial, large-scale production simultaneously, the easy artificial regulatory of working condition and production process, quality is controlled easily; Product can carry out serialization, mass production; The muscardine preparation that particularly uses as biological pesticide can reduce greatly to be produced and use cost, improves the product insecticidal effect, has very high economic worth and good prospects for application.

Four, description of drawings

Fig. 1: muscardine preparation indoor control cabbage caterpillar effect

Five, embodiment

Below in conjunction with instance the present invention is done further description:

Embodiment 1: solid culture method

1, thorough disinfection sterilization, cultivation pool, koji tray, plastics bag, culturing room and cultivation room air.

2, bright silkworm chrysalis, silk mill boiled that silkworm chrysalis behind silk cocoon liquid, the filature, pupa slag or the dried pupa of being processed by silkworm chrysalis, pupa powder, pupa are rotten to add an amount of moisture content; The adjustment water cut reaches about 50%wt; Form solid medium, the solid medium branch is packed in cultivation pool, koji tray or the plastics bag.

3, packed above-mentioned raw materials is placed high-pressure sterilizing pot sterilization 60 minutes, temperature is 125 ℃ in the maintenance Autoclave, and pressure is 0.15MPa.

4, take out sterilized packed substratum, it is indoor to be placed on aseptic inoculation.

5, when substratum in the bag is cooled to 30 ℃, the inoculation muscardine, the weight ratio that inoculum size accounts for substratum is 2～30%, control culturing room temperature is 25-30 ℃.Be placed on cultivation pool or the culturing rack of culturing room and cultivate, cultivate, after inoculation, plastics bag is evenly punched like plastics bag.

6, need repeatedly stir plastics bag in the culturing process, or substratum in koji tray or the stirring cultivation pool, so that substratum is evenly ventilative, muscardine evenly growth makes full use of culture material.

7, plate method can carry out drying, concentrate after 5～7 days, added 5-20% wheat bran, the clay of 10-30% or the product that attapulgite can be processed different physical aspects of culture weight.

Embodiment 2: liquid culture method

1, silkworm chrysalis or pupa ground-slag is broken, process the raw material of powdery or little granulated, preferably particle diameter is below 80 orders.

2, above-mentioned raw materials is dropped in the fermentor tank, add capacity moisture content, making the culture material water cut is 90%wt, and adjustment pH is 6.5-7.2, and keeping the interior temperature of fermentor tank is 121 ℃, and pressure is 0.12MPa, sterilizes 40 minutes.

3, fermentor tank cooling, make a jar interior culture medium temperature be 32 ℃ after, inoculate muscardine, the weight ratio that the muscardine inoculum size accounts for jar interior substratum is 10%.

4, after the inoculation, keeping fermentor tank pressure is 0.1MPa, and in jar, feeds sterile air continuously, and air flow is 0.4VVM.

5, the liquid culture time is about 3～5 days, after the end, can carry out press filtration, concentrates, absorption, evaporation drying, adds the 5-20% wheat bran of culture weight, clay or the attapulgite of 10-30% can be processed solid, powder or liquid product.

Embodiment 3:

1, preparing culture medium: said substratum contains silkworm chrysalis, or silkworm chrysalis and water is by weight the mixture of 20-50: 80-50, or boils silk cocoon liquid, or silkworm chrysalis and boil the mixture of silk cocoon liquid by weight 10-40: 90-60;

2, sterilization: with inserting behind the medium sterilization in the culture vessel, sterilising conditions is 115～130 ℃, 0.1～0.15MPa again, and the time is 30～90 minutes.

3, inoculation culture: substratum is cooled to 30～32 ℃, the inoculation muscardine, and the weight ratio that the muscardine of inoculation accounts for substratum is 2%wt～30%wt; And pass to sterile air, keeping culture temperature is 20～30 ℃, cultivates after 5～7 days; Add the 5-20% wheat bran of total culture weight, clay or the attapulgite of 10-30%; Mix thoroughly, filter press or 35 ℃ of oven dry are pulverized and are processed the solid powdery product.

Embodiment 4: the application on kill pests

Example one: kill cabbage caterpillar

Product muscardine pulvis of the present invention is converted 10 times of dilutions of water, process mother liquor, mixing is subsequent use.Be diluted to 0 respectively then, 100mg/L, 200mg/L, 300mg/L, 500mg/L, 1000mg/L, 2000mg/L is immersed in the above-mentioned solution 30 minutes respectively with the fresh vegetable blade, takes out, and dries 20 minutes.Put into the 1000mL glass beaker that fills 50 cabbage caterpillars, seal, cabbage caterpillar is placed in the illumination box together with beaker, 28 ℃ of daytimes, 18 ℃ of nights, illumination every day 12 hours, humidity about 85% with gauze.Check the cabbage caterpillar death condition every day one time.The high mortality of cabbage caterpillar after 3 days reaches 83.5% (Fig. 1).

Example two: kill Pyrausta nubilalis (Hubern).

Product muscardine pulvis of the present invention is converted 10 times of dilutions of water, process mother liquor, mixing is subsequent use.Be diluted to 0 respectively then, 200mg/L, 400mg/L, 800mg/L, 1600mg/L, 3200mg/L is immersed in the above-mentioned solution 30 minutes respectively with the fresh corn blade, takes out, and dries 20 minutes.Put into the 1000mL glass beaker that fills 50 Pyrausta nubilalis (Hubern).s, seal, Pyrausta nubilalis (Hubern). is placed in the illumination box together with beaker, 30 ℃ of daytimes, 20 ℃ of nights, illumination every day 12 hours, humidity about 75% with gauze.Check the death condition of a Pyrausta nubilalis (Hubern). every day.The high mortality of cabbage caterpillar after 5 days reaches 86.1%, minimumly reaches 32.4%.

Claims

1. produce the method for muscardine, it is characterized in that first preparing culture medium, said substratum is broken with silkworm chrysalis or pupa ground-slag, processes the raw material of powdery or fine granularity, and particle diameter is below 80 orders; Above-mentioned raw materials is dropped in the fermentor tank, add capacity moisture content, making the culture material water cut is 90%wt, and adjustment pH is 6.5-7.2, and keeping the interior temperature of fermentor tank is 121 ℃, and pressure is 0.12MPa, sterilizes 40 minutes; Fermentor tank cooling, make a jar interior culture medium temperature be 32 ℃ after, inoculate muscardine (Beauveria brongniartii), the weight ratio that the muscardine inoculum size accounts for jar interior substratum is 10%; After the inoculation, keeping fermentor tank pressure is 0.1MPa, and in jar, feeds sterile air continuously, and air flow is 0.4VVM; The liquid culture time is 3～5 days, after the end, can carry out press filtration, concentrates, absorption, evaporation drying, adds the 5-20% wheat bran of culture weight, clay or the attapulgite of 10-30% processed solid, powder, liquid product.