CN101560482B - Buprofezin pesticide residual degrading bacteria and produced microbial inoculum thereof - Google Patents
Buprofezin pesticide residual degrading bacteria and produced microbial inoculum thereof Download PDFInfo
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- CN101560482B CN101560482B CN2009100271125A CN200910027112A CN101560482B CN 101560482 B CN101560482 B CN 101560482B CN 2009100271125 A CN2009100271125 A CN 2009100271125A CN 200910027112 A CN200910027112 A CN 200910027112A CN 101560482 B CN101560482 B CN 101560482B
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Abstract
The invention provides a degrading residual microbial inoculum of buprofezin pesticide, belonging to the field of biological high technology. The used bacterial strain is gram staining negative bacteria BF3, and is identified as Paracoccus (Paracoccus sp.). The main biological characteristic is gram-negative and cell is clubbed. Bacterial colony is milky yellowish, round, swollen and opaque, withneat margin and smooth surface. The bacterial strain is facultative anaerobic and chemoheterotropic. The culture solution obtained by fermenting BF3 strain is the microbial inoculum. The application of degrading bacteria product can ensure that buprofezin pesticide residue quantity in soil is reduced by more than 90%, and can solve the overproof problem of buprofezin pesticide residue in agricultural production.
Description
One, technical field
The invention provides the microbial inoculum of a kind of buprofezin pesticide residual degrading bacteria and production thereof, belong to biological high-tech field, is to utilize method of microorganism degraded chemical pesticide residual, is applicable to the production of green non-polluted farm product in the modern agriculture production.
Two, background technology
The use of chemical pesticide has promoted modern agricultural development widely, for the loss that causes because of reasons such as phytopathy, worm, crop smotherings has in a large number been retrieved in agriculture production, and greatly improved the efficiency and the mechanization degree of agriculture production, in the modernization of agriculture production, played immeasurable effect.But agricultural chemicals is some toxic and harmful substances simultaneously, after they enter ecotope, has not only killed the deleterious biology to the mankind, also the ecosystem has been caused serious harm simultaneously.A large amount of uses of agricultural chemicals cause that pesticide residue exceed standard in the agricultural-food, have 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 protection consciousness, the whole society also more and more pays close attention to direct harm and potential impact that pesticide residue are brought, and people press for the cleaning green agricultural product that non agricultural chemical residuum pollutes.But agricultural form and demographic situation from present China, use to stop agricultural chemicals that to reduce farm output be impracticable as the nuisanceless production model of cost, agricultural chemicals also will continue to bring into play enormous function as the effective means of controlling plant disease, worm, crop smothering in agriculture production.How solving this double-barreled question in the agriculture production just becomes the research focus of each subject.Laboratory and real application research all show and utilize degrade pesticide residue in the soil of microorganism, thereby reduce the pesticide residue in the agricultural-food such as grain, vegetables, tealeaves, and the quality and the economic worth that improve agricultural-food are feasible.This seminar has had the patent of invention of degradation bacteria (open day: on March 19th, 1997), this patent was a bacillus J5, degrading methamidophos pesticide residue effectively for ZL96106317.3, publication number: CN1145408A in the prior art.
Three, summary of the invention
Technical problem the objective of the invention is at practical problems in the production practice and demand, develops out a kind of novel bacterium for degradating residual agricultural chemical, uses this microbial inoculum that the residual quantity of Buprofezin is reduced more than 90%, and production and use cost are lower.Use this degradation bacterial agent in the normal growth process of farm crop, normally to use chemical pesticide control and guarantee that the agricultural chemicals residual content meets the green food requirement in the agricultural-food.
Be main contents of the present invention below the technical scheme:
The invention provides a kind of degradation bacterial agent of eliminating buprofezin pesticide residual, bacterial strain uses therefor is Gram-negative bacterium BF3, be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on April 3rd, 2009, culture presevation number is: CGMCC No.3001, and through being accredited as secondary coccus (Paracoccus.sp).Main biological characteristics is a Gram-negative, the cell rod-short.Bacterium colony is milky white yellow partially, circle, and protuberance, opaque, the edge is neat, smooth surface.Amphimicrobian, chemoheterotrophy.
The technology of using above-mentioned buprofezin pesticide residual degrading bacteria to produce microbial inoculum is: test tube slant kind-shake bottle a kinds-seeding tank-production jar-product (pack formulation and be liquid bacterial agent or solid absorption microbial inoculum).Detailed implementation step of the present invention is:
1. buprofezin pesticide residual degrading bacteria BF3 test tube kind is inoculated in broth culture and shakes in the bottle, shaking culture is to logarithmic phase;
2. above-mentioned cultured bacterial classification is inoculated into 500 liters of seeding tanks by 10% inoculum size of seed tank culture base, be cultured to logarithmic phase, the culture medium prescription (mass percent) that seeding tank is used: glucose 0.8%, NH4NO3 1%, K2HPO4 0.2%, MgSO4 0.05%, and NaCl 0.01%, CaCO
30.3%, yeast extract paste 0.02%, pH value 7.2-7.5;
3. seed liquor is produced a jar cultivation by the 10% inoculum size access of production jar substratum, it is identical with the seed tank culture base to produce the used substratum of jar;
The air flow of sterile air is 1 in the culturing process of seeding tank and production jar: 0.6-1.2, and stirring velocity is 180-240 rev/min, and culture temperature is 30-35 ℃, and the whole process flow incubation time is 36-48 hour.Nutrient solution was microbial inoculum after fermentation was finished.
It is low, easy to use that the bacterium for degradating residual agricultural chemical that beneficial effect uses this invention to produce has the production use cost, and the advantage that removal effect is good is adapted at national grain oil ﹠ vegetable production export base or has the regional big area of green food brand mark to promote the use of.The present invention is for preserving the ecological environment, and the protection people health improves additional value of farm products and has great importance.Degradation bacteria BF3 can use this degradation bacterial agent normally to use chemical pesticide to carry out prevention and control of weeds and guarantee that the agricultural chemicals residual content meets the green food requirement in the agricultural-food in the normal growth process of farm crop with buprofezin pesticide residual degrading in the soil more than 90% (table 1).And this bacterium is a Gram-negative bacteria, the effect that agricultural chemicals uses after the survival time in field suitably can not influence.
The present invention has successfully solved in the agriculture production buprofezin pesticide residual problem that exceeds standard, and has both given full play to the efficiently fast effect of chemical pesticide in prevention and control of weeds, can produce non-toxic and non-pollution green agricultural product again.
Four, embodiment
The invention provides a kind of degradation bacterial agent of eliminating buprofezin pesticide residual, bacterial strain uses therefor is Gram-negative bacterium BF3, separates from the Nanjing paddy soil.
Be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on April 3rd, 2009, culture presevation number is: CGMCC No.3001, and through being accredited as secondary coccus (Paracoccus sp.).Main biological characteristics Gram-negative, the cell rod-short.Bacterium colony is milky white yellow partially, circle, and protuberance, opaque, the edge is neat, smooth surface.Amphimicrobian, chemoheterotrophy.
Degradation bacteria BF3 degradation characteristic is measured in the liquid nutrient medium: the single bacterium colony of picking BF3 is in 5ml LB liquid nutrient medium (peptone 10 grams, yeast extract paste 5 grams, NaCl 10 grams, 1000 milliliters in water, pH 7.0～7.2) in, 30 ℃, 170rmp/min shaking culture 24h obtain fresh bacterium liquid.Liquid nutrient medium (the NH that contains the 50mg/L Buprofezin at 100ml
4NO
31.0 gram, K
2HPO
41.5 gram, KH
2PO
40.5 gram, MgSO
40.2 gram, NaCl 0.5 gram, peptone 1 gram, yeast extract paste 0.5 gram, 1000 milliliters in water, pH 7.0～7.2) in, the bacterium liquid after the centrifugal suspension of access 5ml is cultivated (170rmp/min) in 30 ℃ of shaking tables, timing sampling is measured, measure the content of Buprofezin, the results are shown in Table 1, the degradation rate of degradation bacteria BF3 parathiazine ketone reaches more than 90% in the liquid nutrient medium.
Degraded in soil bacterium BF3 parathiazine ketone degradation characteristic is measured: take by weighing 1000g soil, add Buprofezin, make that to contain concentration in the soil be 50mg/kg, add BF3 bacterium liquid 50ml, mixing places constant temperature culture under 30 ℃ of incubator dark conditions, and timing sampling is measured the residual quantity of Buprofezin in soil, the results are shown in Table 1, degraded in soil bacterium degraded Buprofezin reaches more than 90%.
Table 1: the degradation characteristic of bacterial strain parathiazine ketone in inorganic salt liquid substratum sweetening of the soil
| Substratum | Soil | Liquid nutrient medium |
| Degradation rate (%) | ＞90％ | ＞90％ |
The original seed of pesticide residual degrading bacteria BF3 of the present invention is activated on culture dish, and measure degradation property, be inoculated on the test tube slant standby.Inclined-plane kind culture medium prescription: peptone 10 grams, yeast extract paste 5 grams, NaCl 10 grams, 1000 milliliters in water, agar 20 grams, pH 7.0～7.2.
The test tube kind is inoculated in the 1000ml that contains 200ml broth culture (NaCl 5 grams, 1000 milliliters in water, pH 7.0～7.2 for extractum carnis 3 gram, peptone 10 grams) and shakes in the bottle, and constant-temperature shaking culture is prepared the inoculation seeding tank to logarithmic phase.500 liters of seeding tanks, 400 liters of charging capacitys, culture medium prescription (mass percent): glucose 0.8%, NH
4NO
31%, K
2HPO
40.2%, MgSO
40.05%, NaCl 0.01%, CaCO
30.3%, yeast extract paste 0.02%, pH value 7.2-7.5.The back 121 ℃ of high pressure moist heat sterilizations that finish that feed intake, be cooled to 33 ℃ after, the above-mentioned cultured bottle bacterial classification that shakes is inoculated into 500 liters of seeding tanks by 10% inoculum size, be cultured to logarithmic phase, stirring velocity is 220 rev/mins, sterile air feeding amount is 1: 0.8.The seed liquor that arrives logarithmic phase is produced jar by 10% inoculum size access cultivate, it is identical with the seed tank culture base to produce the used medium component of jar.Produce 5 tons of tankages, 4.5 tons of charging capacitys.Production jar 1.1kg/cm after feeding intake
2Pressure under, 121 ℃ of high pressure moist heat sterilizations, below the sterilization postcooling to 35 ℃, logical sterile air keeps sterile state standby.A postvaccinal production jar temperature is controlled at 30-35 ℃, and the air flow of sterile air is 1 in the culturing process of production jar: 0.8-1.0, and stirring velocity is 240 rev/mins, the whole process flow incubation time is 36-48 hour.After the fermentation ends thalline quantity reach 1,000,000,000/more than the ml.Fermentation is finished the back nutrient solution and is gone out jar and directly be distributed into liquid dosage form with plastic barrel or packing bottle or adopt peat to adsorb and be distributed into the solid fungicide formulation with packing bag.
Claims (2)
1. the degradation bacteria BF3 of a buprofezin pesticide residual, be Gram-negative bacteria, through being accredited as secondary coccus (Paracoccus sp.), be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on April 3rd, 2009, culture presevation number is: CGMCC No.3001.
2. bacterium for degradating residual agricultural chemical of producing with the described buprofezin pesticide residual degrading bacteria of claim 1 produces by the following method:
1) the described buprofezin pesticide residual degrading bacteria BF3 of claim 1 bacterial classification inoculation is shaken in the bottle in broth culture, shaking culture is to logarithmic phase;
2) above-mentioned cultured bacterial classification is inoculated into 500 liters of seeding tanks by volume ratio 10% inoculum size of seed tank culture base, be cultured to logarithmic phase, the used mass percent culture medium prescription of seeding tank is: glucose 0.8%, NH
4NO
31%, K
2HPO
40.2%, MgSO
40.05%, NaCl 0.01%, CaCO
30.3%, yeast extract paste 0.02%, pH value 7.2-7.5;
3) seed liquor is produced a jar cultivation by the volume 10% inoculum size access of production jar substratum, it is identical with the seed tank culture base to produce the used substratum of jar;
4) air flow of sterile air is 1 in the culturing process of seeding tank and production jar: 0.6-1.2, stirring velocity is 180-240 rev/min, culture temperature is 30-35 ℃, the whole process incubation time is 36-48 hour, after the fermentation ends thalline quantity reach 1,000,000,000/more than the ml, fermentation is finished the back nutrient solution and is gone out jar and directly be distributed into liquid dosage form with plastic barrel or packing bottle or adopt peat to adsorb and be distributed into the solid fungicide formulation with packing bag.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100271125A CN101560482B (en) | 2009-05-22 | 2009-05-22 | Buprofezin pesticide residual degrading bacteria and produced microbial inoculum thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100271125A CN101560482B (en) | 2009-05-22 | 2009-05-22 | Buprofezin pesticide residual degrading bacteria and produced microbial inoculum thereof |
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| Publication Number | Publication Date |
|---|---|
| CN101560482A CN101560482A (en) | 2009-10-21 |
| CN101560482B true CN101560482B (en) | 2010-11-03 |
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| CN2009100271125A Expired - Fee Related CN101560482B (en) | 2009-05-22 | 2009-05-22 | Buprofezin pesticide residual degrading bacteria and produced microbial inoculum thereof |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103114065B (en) * | 2013-03-15 | 2014-11-05 | 南京农业大学 | Piperazine degraded paracoccus PQ-01 and application thereof |
| CN107446850A (en) * | 2017-08-23 | 2017-12-08 | 江苏南资环保股份有限公司 | One plant of Buprofezin degradation bacteria and its application |
| CN107699523B (en) * | 2017-11-03 | 2020-06-05 | 南京农业大学 | Bacterium capable of degrading pesticides buprofezin and bifenthrin and microbial inoculum produced by bacterium |
| CN110255717B (en) * | 2019-06-06 | 2020-12-11 | 华南农业大学 | Degrading bacterium of strobilurin fungicide and application thereof |
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2009
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