CN1055119C - Chemical residual degradation vaccine and its prodn. method - Google Patents

Abstract

The present invention provides a pesticide residue degrading microbial inoculum. The bacterial strain used is a bacterial strain which is named as J5 and is in bacillus (class number: bacillus sp.). The bacterial strain has the main characteristic of G<+>; the individual is in a long rod shape; the size is from 1 to 2*3 to 4cm; the bacterial strain has flagellums on the surface; the juvenile culture object is in a long chain shape; the old culture object forms elliptical spores. The production technology comprises seeding liquid, seeding pots, production pots and products. Indicated in tests, the direct application of the product on crops can make the residue removing rate of methamidophos, etc. reach more than 95 %. By using the present invention, chemical pesticides still can be normally used in the process of producing foodstuff and vegetables without pesticide residues, and foodstuff and vegetables which are qualified and have no pesticide residues can be produced.

Description

A kind of bacterium for degradating residual agricultural chemical and production method thereof

The invention belongs to biotechnology, is to utilize the microbiological deterioration pesticide residue, is applicable to the production of non agricultural chemical residuum grain in the agriculture production.

Widely used chemical pesticide in the agriculture production has not only caused the severe contamination of environment, and has caused exceeding standard of pesticide residue in the agricultural-food, has had a strong impact on the foreign exchange earning of people's healthy and agricultural-food.Along with the raising of people's living standard and the enhancing of environmental consciousness, people press for the agricultural-food of non agricultural chemical residuum, if but in agriculture production, do not use chemical pesticide, because the invasion and attack of disease and pest, to cause the underproduction of farm crop inevitably, this is unpractical to the China that will solve 1,200,000,000 populations and have a meal.Chemical pesticide also will continue to use as the effective means of control disease and pest.In order to overcome the above problems, scientist has carried out a large amount of tests, but mostly concentrate on the chemical substance of seeking a kind of " residual few; as to decompose fast; the drug effect height ", and utilizing pesticide residual degrading bacteria to eliminate the pesticide residue in the soil, the pesticide residue in the reduction agricultural-food, the grain and the vegetables that produce nothing or low pesticide residue yet there are no report.

The present invention is exactly at above-mentioned production practical situation, develops a kind of bacterium for degradating residual agricultural chemical, makes chemical pesticide residue degradings such as acephatemet more than 90%.Use this technology in process of crop growth, can normally use chemical pesticide, thereby destroying plant diseases and insect pests guarantees crop yield, can make in grain and the vegetables pesticide residue very low or do not have simultaneously again.

Narrate main contents of the present invention below:

Representative microbial provided by the present invention is the strain (systematic name of bacillus

Bacillus sp.) called after J ₅Bacterial strain (be deposited at Chinese microorganism strain on February 8th, 1996 and preserved management committee common micro-organisms center, CGMCCNO.0252).This bacterial classification principal character is: G ⁺, individual morphology is an elongated rod shape, size is 1-2 * 3-4cm, and the tool peritrichous, young culture is generally long-chain, and old culture can form gemma, and the gemma ellipse is in cell central authorities and littler than cell.Optimum growth temperature is 30-35 ℃, and the suitableeest growth PH is 7.0-7.5, the property of being sick of of holding concurrently.This bacterial strain can be cultivated with the general fermentor tank in the general fermentation industry.

The production technique that the present invention adopts is seed liquor → seeding tank → production jar → product packing (or with packing again after the adsorbents adsorb).

Detailed implementation step of the present invention is:

1) with bacterial classification inoculation of the present invention in broth culture, be cultured to logarithmic phase;

2) above-mentioned cultivation bacterial classification is inserted seeding tank, be cultured to logarithmic phase.The substratum of seeding tank is: sucrose or glucose 0.6-1.0%, (NH ₄) ₂SO ₄0.05-0.20%, K ₂HPO ₄0.1-0.35%, MgSO ₄0.02-0.06%, NaCl0.01-0.02%, CaCO ₃0.05-0.20%, yeast extract paste 0.02-0.05%;

3) seed liquor is inserted production jar cultivation, producing a jar substratum is sucrose 0.1-0.3%, starch 0.5-0.8%, and all the other compositions and content are with the seeding tank substratum;

In culturing process, feed sterile air, air flow is 1: 0.6-1: 1.2, stirring the speed of trembling is 180-240 rev/min, and culture temperature is 30-35 ℃, and incubation time is 24-30 hour, but the nutrient solution direct packaging becomes the also available sorbent material of product, as carrying out packing again after the peat absorption.

The present invention is used for agriculture production can remove pesticide residue effectively, and it is easy to use, and cost is low, and is effective.Bacterium for degradating residual agricultural chemical directly is sprayed in the soil or plant surface after crop is used agricultural chemicals, and it is pesticide residue such as degrading methamidophos rapidly, and the pesticide residue clearance reaches more than 95%.

This invention reduces the quality of pesticide residue raising agricultural-food in the agricultural-food for the protection environment, and protection people's health has great importance.Large scale application is carried out in the place that is adapted at national grain vegetables export base or has a green food logo.

Below narrate embodiments of the invention.

Embodiment 1

Slant strains is seeded in the broth culture, is cultured to the logarithmic growth after date, insert seeding tank again.Seeding tank is 0.5 ton, and charging capacity is 0.4 ton, and medium component is sucrose 3.2kg, (NH ₄) ₂SO ₄0.4kg, K ₂HPO ₄0.8kg, MgSO ₄0.2kg, NaCl0.04kg, CaCO ₃, 0.4kg, yeast extract paste 0.16Kg, seeding tank palpus elder generation is with steam sterilizing and be cooled to 30-35 ℃, and inoculum size is 1%, and the seed tank culture temperature is controlled at 32-33 ℃, stirring velocity is 220 rev/mins, and sterile air feeding amount is 1: 0.8, is cultured to the logarithmic growth after date, seed liquor is inserted the production jar, producing jar is 5 tons, 4.5 tons of charging capacitys, and medium component is sucrose 7.8kg, starch 30kg, (NH ₄) ₂SO ₄4.5kg, K ₂HPO ₄9.0kg, MgSO ₄2.3kg, NaCl0.9kg, CaCO ₃4.5kg yeast extract paste 1kg produces jar in advance with steam sterilizing and be cooled to 30-35 ℃, producing a jar stirring velocity is 240 rev/mins, and air flow is 1: 0.8-1, and to cultivate and finish that thalline quantity reaches more than 1,500,000,000/ml in the nutrient solution of back, nutrient solution is product.

Embodiment 2

With the nutrient solution of embodiment 1 gained, with peat absorption, the ratio of nutrient solution and peat is 1: 3, packs behind the stirring and evenly mixing and is product.

After paddy rice sprayed acephatemet, the bacterium for degradating residual agricultural chemical of above-mentioned production is applied to rice terrace (2kg/ mu), treat rice harves after, with methamidophos residue content in the gas Chromatographic Determination rice, (test of green food test center of Agricultural University Of Nanjing) the results are shown in Table 1.Can find out that from table the methylamine phosphate content comparison of using in its rice of bacterium for degradating residual agricultural chemical of the present invention is according to low more than 95% in the rice.

Annotate: sample sequence number 1,2,3,4,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25 is for using the rice sample of bacterium for degradating residual agricultural chemical of the present invention in the table 1, and sample sequence number 5,26 is contrast rice sample.Table 1

The sample sequence number	Instrument condition	Test result	Remarks
				1	FPD	0.001	mg/kg
2	FPD	ND	mg/kg
				3	FPD	ND	mg/kg
4	FPD	ND	mg/kg
				5ck	FPD	0.007	mg/kg
6	FPD	0.001	mg/kg
				7	FPD	0.001	mg/kg
8	FPD	0.002	mg/kg
				9	FPD	0.003	mg/kg
10	FPD	ND	mg/kg
				15	FPD	ND	mg/kg
19	FPD	ND	mg/kg
				20	FPD	ND	mg/kg
21	FPD	ND	mg/kg
				22	FPD	0.002	mg/kg
23	FPD	0.002	mg/kg
				25	FPD	ND	mg/kg
26ck	FPD	0.016	mg/kg

Claims (2)

1, a kind of microorganism that is used for chemical residual degradation is characterized in that this microorganism is the J of bacillus (Bacillus sp.) ₅Bacterial strain, preserving number CGMCCNO.0252.

2, a kind of method of utilizing the described microorganisms producing bacterium for degradating residual agricultural chemical of claim 1 is characterized in that:

1) with bacterial classification inoculation in broth culture, be cultured to logarithmic phase;

2) above-mentioned cultivation bacterial classification is inserted seeding tank, be cultured to logarithmic phase, the substratum of seeding tank is: sucrose or glucose 0.6-1.0%, (NH ₄) ₂SO ₄0.05-0.20%, K ₂HPO ₄0.1-0.35%, MgSO ₄0.02-0.06%, NaCl0.01-0.02%, CaCO ₃0.05-0.20%, yeast extract paste 0.02-0.05%;

3) seed liquor is inserted production jar cultivation, producing a jar substratum is sucrose 0.1-0.3%, starch 0.5-0.8%, and all the other compositions and content are with the seeding tank substratum;

4) in culturing process, the sterile air air flow is 1: 0.6-1.2, and stirring velocity is 180-240 rev/min, and culture temperature is 30-35 ℃, and incubation time is 24-30 hour.