CN1276713C - Microbial pesticide and its prepn. method - Google Patents

Abstract

The present invention relates to a microbial pesticide and a preparing method thereof. The microbial pesticide is characterized in that the activity strain of the microbial pesticide is the new strain Ba9808-2-3 of Bacillus thuringiensis subspecies. The preparing method of the microbial pesticide comprises the following steps: (1) in the production of the strain, the microbial strain used by the present invention is the new strain Ba9808-2-3 of Bacillus thuringiensis subspecies; (2) in the culture of a culture medium, a nutrient source adopts a carbon source and a nitrogen source, wherein the carbon source can be starch, dextrin, dextrose, maltose, etc., the nitrogen source can be peptone, soybean cake powder, beef extract, corn steep liquid, etc., and other nutrient components, such as salt, phosphate, potassium salt, calcium salt, etc., are added on the basis of the nutrient source; (3) the use of a cultural method, the microbial pesticide can be cultured by a liquid submerged fermentation method, culture temperature is from 27 to 30 DEG C, pH is from 7 to 9, and culture time is from 25 to 40 hours; (4) in the after-treatment of a product, culture liquid is modulated into the liquid with a certain amount of spore, a right quantity of preservative agents, a right quantity of emulsifying agents, a right quantity of ultraviolet protective agents, etc. are added, and then the product is obtained. The present invention is an agricultural insecticide without nuisance, and the present invention has particular effect on important injurious insects for the production of vegetables, such as alkekengi ladybird beetles, striped flea-beetles, daicon leaf beetles, etc.

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 (Bacillusthuringiensis 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 thuringiensis 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:

For the examination insect	The handling dead borer population	Lethality/%	Contrast dead borer population	Lethality/%	Corrected mortality
						Chrysomelidae Chrysomelidae daicon leaf beetle striped flea-beetle	45 45	100 100	3 1	6.7 2.2	100 100
Culculionidae Curculionidae rice water weevil corn weevil	16 16	33.3 35.6	5 2	11.1 4.4	25.0 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

Handle	Extension rate	Insect population radix before the medicine/10 strains			Insect population radix after 7 days/10 strains			Control efficiency
									Larva	Ovum	Adult	Larva	Ovum	Adult
		This microbial pesticide	1∶200	20	15	5	7								5	4	60.0％
Phoxim	1∶3000							28	9	5	8	5	2	64.3％
		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 Bacillus of bacillus thuringiensis Mo Lixun subspecies thuringiensis subsp.Morrisoni Ba9808-2-3, CGMCC No.0920.

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

Adopt the new bacterial strain Bacillus of bacillus thuringiensis Mo Lixun subspecies thuringiensis subsp.Morrisoni Ba9808-2-3, CGMCC No.0920;

(2) medium

Nutrient source in the medium adopts carbon source and nitrogenous source, and wherein carbon source is starch, dextrin, glucose or maltose; Nitrogenous source is peptone, soybean cake powder, beef extract or corn steep liquor, adds salt, phosphate, sylvite or calcium salt on this basis again;

(3) cultural method

Cultivate by liquid submerged fermentation method, cultivation temperature is 27～30 ℃, PH7～9, and incubation time is 25～40 hours;

(4) product post processing

After the culture fluid modulation, add preservative, emulsifier and uv-protector again, be product.

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