CN1513327A - Microbial pesticide and its prepn. method - Google Patents

Abstract

A microbial agricultural chemical for vegetables is prepared from the Bacillus thuringiensis subspecies Ba9808-2-3 through preparing culture meidum from carbon source (starch, dextrin, glucose, etc), nitrogen source (peptone, soybean cake powder, beef extract, etc), edible salt, phosphate, K salt and Ca salt, deep fermenting culture in liquid at 27-30 deg.C pH 7-9 for 25-40 hr, and post treatment. Its advantages are high effect and no environmental pollution.

Description

A kind of microbial pesticide and preparation method thereof

Technical field

The invention belongs to the microbial fermentation engineering technical field, relate to a kind of microbial pesticide and production method thereof.

Background technology

Damage by disease and insect brings about great losses to agricultural production every year, and in order to realize high and stable yields, people mainly adopt chemical pesticide to control always.Chemical pesticide has advantages such as preventive effect height, speed is fast, the insecticidal and antibacterial spectrum is wide, cost is low, use is simple, thereby has obtained developing rapidly, and becomes the important means that the agricultural production good harvest is captured in prevention and elimination of disease and pests.According to statistics, the chemical control area accounts for the ratio of total control area up to about 95% in the period of China 1985～2002.Because using for a long time, in a large number and repeatedly of chemical pesticide brought the pollution to soil, water body and atmosphere; Residue of pesticide increase in the agricultural byproducts, have also directly endangered human beings'health and existence; Simultaneously, the pesticide resistance of insect and germ is constantly deepened and is enlarged, and the existing at least 504 kinds of insects in the whole world have produced resistance to agricultural chemicals, and the pest resistance that has is up to several ten thousand times; In addition, chemical pesticide has has also killed and wounded natural enemy and other beneficial organism in kill pests, destroyed the ecological balance, causes that insect is rampant more.Like this, have to strengthen the amount of application and the frequency of agricultural chemicals, thus the vicious circle that causes chemical pesticide to use.For man-land relationship area at full stretch, its only way is to greatly develop biopesticide.

Summary of the invention

The present invention seeks to overcome the shortcoming of existing agricultural chemicals, a kind of microbial pesticide and production method thereof are provided, this agricultural chemicals has efficient, safe, nuisanceless and characteristics such as production cost is low.

This microbial pesticide, its active bacterial strain (effective ingredient) is the new bacterial strain Ba9808-2-3 of bacillus thuringiensis Mo Lixun subspecies (Bacillustharingensis subsp.morrisoni), and controlling object is coleoptera Coccinellidae insect Henosepilachna vigintioctopunctata, Chrysomelidae insect daicon leaf beetle and striped flea-beetle etc.

The preparation method of this microbial pesticide is as follows:

1. active bacterial strain is identified

Bacterial strain Ba9808-2-3 serotype is H _8a8b.Identify that by morphological feature trophosome is the direct rod shape somatic cells that can produce gemma, bacterium is held garden slightly, 1.1～1.5 microns * 2.8～4.5 microns of sizes, thalline whole body flagellum, motion, Gram-positive.Identify contrast through biochemical reaction mensuration, flagellar antigen analysis, somatic antigen analysis, the analysis of esterase electrophoretogram etc., all similar to bacillus thuringiensis Mo Lixun subspecies.But at aspects such as some morphological features obvious difference is arranged again, in the growth and breeding process, parasporal crystal is difficult for seeing when forming in sporangium, is key character of this bacterial strain; And the parasporal crystal after discharging is irregular, nearly little square, 0.5～0.6 * 0.7 micron of size; Colonial morphology, milky, little, round, the edge is smooth.The poisoning object of this bacterial strain also has specific characteristics, and is mainly coleoptera Coccinellidae insect Henosepilachna vigintioctopunctata, Chrysomelidae insect daicon leaf beetle and striped flea-beetle etc.So the Ba9808-2-3 bacterial strain is bacillus thuringiensis Mo Lixun subspecies (Bacillus tharingensis subsp.morrisoni) bacterial strains newly.

2. preparation technology

(1) produces bacterial classification

The used microorganism fungus kind of the present invention is the new bacterial strain Ba9808-2-3 of bacillus thuringiensis Mo Lixun subspecies, is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, and preserving number CGMCC No0920, preservation date are on April 6th, 2003.

(2) medium

Nutrient source in the medium adopts carbon source and nitrogenous source.Wherein carbon source can be starch, dextrin, glucose, maltose etc.; Nitrogenous source can be peptone, soybean cake powder, beef extract, corn steep liquor etc.Add some other nutritions on this basis again, as salt, phosphate, sylvite, calcium salt etc.

(3) cultural method

Microbial pesticide of the present invention can be cultivated by liquid submerged fermentation method.Cultivation temperature is 27～30 ℃, PH7～9, and incubation time is 25～40 hours.

(4) product post processing

Culture fluid is modulated into certain spore content, adds an amount of preservative, emulsifier and uv-protector etc. again and is product.

Figure of description is seen in concrete technological process.

3. insecticidal spectrum

Product is diluted to spore content 10 ⁶The solution of cfu/ml carries out the insecticidal spectrum test, the results are shown in Table 1.As can be seen from Table 1, the low instar larvae to important coleopteran pest daicon leaf beetle, Henosepilachna vigintioctopunctata and striped flea-beetle on the vegetables has strong toxicity, employing 10 ⁶The solution of cfu/ml, the post-equalization lethality was 100% in three days.

Table 1 is that the insecticidal spectrum of the dense medicine of this microorganism is measured:

Handling contrast for examination proofreaies and correct dead

The dead worm lethality of the dead worm lethality of insect rate

Number/% number/%

Chrysomelidae Chrysomelidae

Daicon leaf beetle 45 100 3 6.7 100

Striped flea-beetle 45 100 1 2.2 100

Culculionidae Curculionidae

Rice water weevil 16 33.3 5 11.1 25.0

Corn weevil 16 35.6 2 4.4 32.6

Coccinellidae Coccinellidae

Henosepilachna vigintioctopunctata 45 100 4 8.9 100

Bostrichidae Bostrychidae

Lesser grain borer 19 42.2 5 11.1 35.0

Tenebrionidae Tenebrionidae

Red flour beetle 18 40.0 3 6.7 35.7

Annotate: be 45 for processing of examination borer population and contrast

Advantage of the present invention

1. special efficacy

Early stage sporeine preparation major part is to kill lepidoptera pest, report and developed some in recent years and kill the preparation of other insects such as coleoptera, but have supervirulent sporeine preparation not to be reported Henosepilachna vigintioctopunctata and these two kinds of vegetables important pests of striped flea-beetle.

2. safety

Insecticidal action realizes that by crystalline protein this albumen is toxogen, avirulence own, digested by the strong basicity in the insect gut (PH9-10) digestive juice earlier after searching for food, open crystal structure, be decomposed to form toxalbumin (phallotoxins) by the activating enzyme in the enteron aisle then, act on midgut epithelial cells.People and mammal digestive juice are not had a strong basicity, and the selectivity activating enzyme that natural enemy and aquatic animal do not have insect to have all can not make the crystalline protein activation.So said preparation is very safe to people, animal, insect natural enemy and aquatic animal.

3. inexpensive

Produce bacterium sporulation quantity height, raw materials for production are common agricultural byproducts, be simple and easy to, be suitable for large-scale industrial production, and production equipment can be general with other microniological proudcts, thereby cost is lower.

Description of drawings

Accompanying drawing is a process flow diagram

Embodiment

Embodiment one: the preparation of this pesticidal preparations of spore content 10,000,000,000/ml

In the 70L fermentation tank, be made into the 50L medium by soybean cake powder 2.5%, sweet potato starch rust 1.0%, glucose 1.0%, magnesium sulfate 0.05%, calcium carbonate 0.5%.Be cooled to 30 ℃ after the sterilization according to a conventional method,,, promptly get the zymotic fluid of spore content 6,200,000,000/ml through 30 ℃ ± 1 aerobic culture 30 hours by 5% ratio inoculation production bacterial classification.This zymotic fluid adds 0.1% Sodium Benzoate again and makes preservative through precipitating the suspension that is diluted to spore content 10,000,000,000/ml after concentrating, and 10% saponin makes emulsifier, promptly gets product.This product spore content＞10,000,000,000/ml, suspensibility 60-90%, PH4.5-6.0, fineness is 95% mistake, 100 mesh sieves, spore is brilliant in 1: 0.5.

Embodiment two: use this microbial pesticide preparation control eggplant Henosepilachna vigintioctopunctata application test

Test is carried out in the Ningbo of Zhejiang somewhere, adopts 200 times of dilutions of its 10,000,000,000/ml preparation, and the chemical pesticide contrast is 3000 times of dilutions of 50% phoxim emulsifiable concentrate, and other establishes the clear water blank, totally 3 processing.Every processing area 15m ², each repeats 4 times, randomized arrangement.Investigation in 7 days behind the spray medicine, 10 points are looked in every processing, look into 10 strains at every, and statistics insect population radix calculates control efficiency, the results are shown in Table 2.As shown in Table 2,200 times of dilutions of this preparation of 10,000,000,000/ml are 60.0% to the field control effect of Henosepilachna vigintioctopunctata, and are close with 3000 times of dilution effects of chemical pesticide phoxim emulsifiable concentrate.

Table 2 is the field control effect of this microbial pesticide preparation to the eggplant Henosepilachna vigintioctopunctata

Insect population radix before the diluent/10 strains insect population radix after 7 days/10 strains

Handle control efficiency

Multiple larva ovum adult larva ovum adult

The farming of this microorganism

1∶200 20 15 5 7 5 4 60.0％

Medicine

1∶3000 28 9 5 8 5 2 64.3％

Phoxim

Clear water/45 16 2 45 0 10/

Claims (2)

1. a microbial pesticide is characterized in that active bacterial strain is the new bacterial strain Ba9808-2-3 of bacillus thuringiensis Mo Lixun subspecies.

2. a preparation method who is exclusively used in the described microbial pesticide of claim 1 adopts microbial fermentation technology production, and its preparation technology's characterization step is as follows:

(1) produces bacterial classification

The used microorganism fungus kind of the present invention is the new bacterial strain Ba9808-2-3 of bacillus thuringiensis Mo Lixun subspecies;

(2) medium

Nutrient source in the medium adopts carbon source and nitrogenous source, and wherein carbon source can be starch, dextrin, glucose, maltose etc.; Nitrogenous source can be peptone, soybean cake powder, beef extract, corn steep liquor etc., adds some other nutritions on this basis again, as salt, phosphate, sylvite, calcium salt etc.;

(3) cultural method

Microbial pesticide of the present invention can be cultivated by liquid submerged fermentation method, and cultivation temperature is 27～30 ℃, PH7～9, and incubation time is 25～40 hours;

(4) product post processing

Culture fluid is modulated into certain spore content, adds an amount of preservative, emulsifier and uv-protector etc. again and is product.