CN104988094A - Method for manufacturing quinclorac solid degrading inoculant - Google Patents

Method for manufacturing quinclorac solid degrading inoculant Download PDF

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Publication number
CN104988094A
CN104988094A CN201510384248.7A CN201510384248A CN104988094A CN 104988094 A CN104988094 A CN 104988094A CN 201510384248 A CN201510384248 A CN 201510384248A CN 104988094 A CN104988094 A CN 104988094A
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China
Prior art keywords
solid
quinclorac
inoculant
liquid seeds
making method
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Pending
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CN201510384248.7A
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Chinese (zh)
Inventor
徐淑霞
张世敏
梁振普
高玉千
吴坤
赵培
贺亚斐
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Henan Agricultural University
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Henan Agricultural University
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Priority to CN201510384248.7A priority Critical patent/CN104988094A/en
Publication of CN104988094A publication Critical patent/CN104988094A/en
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Abstract

The invention provides a method for manufacturing a quinclorac solid degrading inoculant, and belongs to the technical field of microorganisms. The method includes the steps that 1, preparation, pH adjustment and sterilization of a medium are conducted; 2, bordetella preserved on a bevel is inoculated, and shake cultivation is carried out to obtain liquid seeds; 3, the liquid seeds are inoculated in a culture medium containing wheat bran, corn straw powder and activated carbon, and are cultured in a constant-temperature culture box in a sealed mode to obtain a target product. The organophosphorus pesticide degrading inoculant produced through the technology has the advantages of being low in product cost and environmentally friendly, and the inoculant can efficiently degrade organophosphorus pesticide residues on the surfaces of plants and in soil. Besides, physical and chemical properties and biological characters of the soil can be improved, soil fertility is improved, immunity of crops can be enhanced, intrusion of bacteria is prevented, and the inoculant particularly has a good prevention function on soil-borne diseases and the continuous cropping barrier. Thus, crop growth is promoted, the agricultural yield is increased, quality of agricultural products is improved, and production cost is reduced.

Description

The making method of the solid-state degradation bacterial agent of a kind of quinclorac
Technical field
The invention belongs to microorganism field, be specifically related to the making method of the solid-state degradation bacterial agent of a kind of quinclorac.
Background technology
Quinclorac belongs to the stronger weedicide of the quinoline acids selectivity of hormone-type.Be the novel hormonal type quinoline carboxylic acid weedicide of early 1980s BASF Aktiengesellschaft's exploitation, be mainly used in the anti-barnyard grass in rice field.Rice field is used quinclorac and is also caused certain harm to the vega of rear stubble, and make cigarette strain grow deformity, plant change is short, new leaf back turnup, shrinkage, leaf is wide seriously suppressed.Agricultural chemicals, as the biological exogenous material of synthetic, is not easily degraded by microorganisms usually, very long in the occurring in nature transformation period, therefore is referred to as intractable compound.But be through long-term natural selection, have a lot of microorganism can degrading pesticide.Microbiological deterioration quinclorac is utilized to be the new technology for curbing environmental pollution grown up nearly ten years.At present, the research for Degradation of Pesticides By Microorganisms mainly concentrates on bacterium.The essence of degradation by bacteria agricultural chemicals is enzymatic reaction, namely compound enters in bacterial body by certain mode, then under various enzyme effect, through a series of biochemical reactions, agricultural chemicals is degradable or resolve into the process of the nontoxic of molecular weight or the less compound of toxicity the most at last.
Summary of the invention
The object of the present invention is to provide the making method of the solid-state degradation bacterial agent of a kind of quinclorac.
Based on above-mentioned purpose, this invention takes following technical scheme:
A making method for the solid-state degradation bacterial agent of quinclorac, comprises the steps:
(1) preparation of solid medium: wheat bran, maize straw and gac prepare in proportion, then adds the distilled water of total raw material two volumes, obtains solid medium after high-temperature sterilization;
(2) pre-treatment of solid medium: regulate pH to be 7.0 with the NaOH aqueous solution, then high-temperature sterilization;
(3) preparation of liquid seeds: the Bordetella bacterial strain HN36 inoculating slant preservation in the triangular flask that LB liquid nutrient medium is housed, in 35 DEG C of shaking tables vibrations, obtains liquid seeds.
(4) be inoculated into by liquid seeds in step (2) pretreated solid medium, mixing, to constant incubator, airtight cultivation obtains solid-state microbial inoculum, and air seasoning, to microbial inoculum water content less than 30%, grinds the rear 0.45mm of mistake and sieves, with for subsequent use.
Preferably, in step (1), wheat bran, maize straw and gac are prepared burden in the percent mass ratio of 85%:10%:5%; In step (2), the concentration of the NaOH aqueous solution is 6mol/L; In step (3), shaking table vibration refers to 150 revs/min of shaking culture 36 hours; Inoculation described in step (4) refers in solid medium, inoculation 10v%(volume percent) liquid seeds; In described constant incubator, airtight cultivation refers to and cultivate 12-48 hour in 35 DEG C of electric heating incubator, and the viable count detected in solid culture matrix reaches 5.0 × 10 8individual/more than g.
The using method of this microbial inoculum can adopt to spread manuer in holes or spreads fertilizer over the fields after cigarette strain is transplanted, and amount of application is 45 ~ 55 kg/ mus.
The invention provides the preparation method of the solid-state microbial inoculum of a strain Bordetella, this microbial inoculum can be used for the quinclorac of degrading in soil.From the waste water from pesticide factory lignin-sludge producing quinclorac, separation and purification is to the bacterial strain HN36 of a high-efficiency degradation quinclorac.The test of combining form, physiological and biochemical property, 16S rDNA sequence and bacterial strain phyletic evolution developmental analysis, bacterial strain HN36 is accredited as Bordetella bacterial strain bordetellasp .degradation characteristic research under shake flask culture conditions shows: HN36 can be sole carbon source and nitrogenous source growth with quinclorac, it is 35 DEG C in optimum temperuture, when pH is 7,48 h are to the separation of the degradation rate of 400 mg/L quincloracs up to this microbial inoculum of 96.2%(, and qualification is specifically see the separation of quinclorac degradation bacteria HN36, qualification and degradation characteristic research volume the 2nd phase 45-49 page in " safety and environment journal " April the 12nd in 2012).
The organophosphorus pesticide degradation bacterium agent that the method is produced, there is low, the eco-friendly feature of production cost, organophosphorus pesticide in product energy efficient degradation plant surface and soil, but also physics and chemistry, the biological character of soil can be improved, increase soil fertility, the immunological competence of crop can be strengthened, prevent the intrusion of germ, especially have good prophylactic effect to soil-borne disease and continuous cropping obstacle.Thus promotion plant growth, boost agricultural yield, improve quality of agricultural product, and save production cost.
Embodiment
Below in conjunction with specific embodiment, technical scheme of the present invention is described in further detail, but protection scope of the present invention is not limited thereto.
Embodiment 1
A making method for the solid-state degradation bacterial agent of quinclorac, comprises the steps:
(1) preparation of solid medium: prepare in the percent mass ratio of 85%:10%:5% with wheat bran, maize straw powder (20-30mm), gac, then add the distilled water of total raw material two volumes, obtain solid medium after 121 DEG C of sterilizing 20min;
(2) pre-treatment of solid medium: regulate pH to be 7.0, in 121 DEG C of sterilizing 20min with the NaOH aqueous solution;
(3) preparation of liquid seeds: the Bordetella bacterial strain HN36 inoculating slant preservation in the triangular flask that LB liquid nutrient medium is housed, in 35 DEG C, 150 revs/min of shaking culture 36 hours (spectrophotometer detects OD value about 1.0), obtain liquid seeds;
(4) in solid medium, by liquid seeds by 10v%(volume percent) be inoculated in step (2) pretreated solid medium, mixing, cultivates 48 hours to 35 DEG C of electric heating incubators, and the viable count detected in solid culture matrix reaches 5.0 × 10 8individual/more than g, air seasoning, to microbial inoculum water content less than 30%, grinds the rear 0.45mm of mistake and sieves, packaging, leaves shady and cool dry place in, use in 12 months.
test example 1
Organophosphorus pesticide degradation bacterium agent is to the degradation effect of tobacco blade face quinclorac pesticide residue
Choose two identical each 30m in tobacco plot 2, evenly spray the quinclorac powder pesticide (in agricultural chemicals, quinclorac effective constituent is 50wt%) of 1000 times of dilutions.After 6h, plot is evenly sprinkled into the solid-state degradation bacterial agent of quinclorac of the present invention 1 kilogram, and another plot as a control group, does not use whatever, turns over evenly; After 36d, five point samplings detect the residual of tobacco blade face quinclorac, find treatment group comparatively control group quinclorac residual quantity minimizing about 40%.
test example 2
The solid-state degradation bacterial agent of quinclorac of the present invention promotes tobacco growing
Adopt the experiment of green house of vegetables community, 8 identical hillslope processes of condition are set, each community 30m 2, 2 communities are 1 group of parallel test, are divided into into 4 groups, plantation tobacco, and the tobacco seedling phase, does not apply fertilizer and compare in the 1st group of 2 communities; 2nd group of 2 community respectively impose the solid-state degradation bacterial agent of quinclorac of the present invention once according to the consumption of 50 kgs/acre after tobacco complete germination He after 3 weeks; Urea is respectively used once according to the rate of fertilizer application of 15 kgs/acre in 3rd group of 2 community after tobacco complete germination He after 3 weeks; Urea is respectively used once according to the rate of fertilizer application of 15 kgs/acre in 4th group of 2 community after tobacco complete germination He after 3 weeks, and 1 day after using urea imposes the solid-state degradation bacterial agent of quinclorac of invention once, the irrigation that each community is identical and way to manage according to the rate of fertilizer application of 50 kgs/acre at every turn.Tobacco growing 5 weeks " Invest, Then Investigate " yield and qualities.Use separately microbiobacterial agent of the present invention and use separately the tobacco production of urea basically identical, all than the check plot volume increase about 12% of not applying fertilizer; The community that microbial inoculum and urea are all used is than check plot volume increase about 18%; In tobacco, NO3-N and NO2-N content is not to apply fertilizer and to use the minimum of microbiobacterial agent separately, at less than 1/3 of national standard Limited Doses, use again after using urea microbiobacterial agent in less than 1/2 of national standard limitation, and use separately NO3-N and NO2-N content in the tobacco of urea and all exceed standard seriously.Show that this microbiobacterial agent has good effect of increasing production, and the quality of tobacco can be significantly improved.

Claims (5)

1. a making method for the solid-state degradation bacterial agent of quinclorac, is characterized in that, comprise the steps:
(1) preparation of solid medium: wheat bran, maize straw and gac prepare in proportion, then adds the distilled water of total raw material two volumes, obtains solid medium after high-temperature sterilization;
(2) pre-treatment of solid medium: regulate pH to be neutral with the NaOH aqueous solution, then high-temperature sterilization;
(3) preparation of liquid seeds: the Bordetella bacterial strain inoculating slant preservation in the triangular flask that LB liquid nutrient medium is housed, in 35 DEG C of shaking tables vibrations, obtains liquid seeds;
(4) be inoculated into by liquid seeds in step (2) pretreated solid medium, mixing, to constant incubator, airtight cultivation obtains solid-state microbial inoculum, and air seasoning, to microbial inoculum water content less than 30%, grinds the rear 0.45mm of mistake and sieves, with for subsequent use.
2. the making method of the solid-state degradation bacterial agent of quinclorac according to claim 1, is characterized in that, in step (1), wheat bran, maize straw and gac are prepared burden in the percent mass ratio of 85%:10%:5%.
3. the making method of the solid-state degradation bacterial agent of quinclorac according to claim 1, is characterized in that, in step (2), the concentration of the NaOH aqueous solution is 6mol/L.
4. the making method of the solid-state degradation bacterial agent of quinclorac according to claim 1, is characterized in that, in step (3), shaking table vibration refers to 150 revs/min of shaking culture 36 hours.
5. the making method of the solid-state degradation bacterial agent of quinclorac according to claim 1, is characterized in that, the inoculation described in step (4) refers in solid medium, the liquid seeds of inoculation 10v%; In described constant incubator, airtight cultivation refers to and cultivate 12-48 hour in 35 DEG C of electric heating incubator, and the viable count detected in solid culture matrix reaches 5.0 × 10 8individual/more than g.
CN201510384248.7A 2015-06-30 2015-06-30 Method for manufacturing quinclorac solid degrading inoculant Pending CN104988094A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106148248A (en) * 2016-09-06 2016-11-23 山东农业大学 A kind of Camellia sinensis special organophosphor decomposer YP8 and bacterial preparation process thereof
CN106305178A (en) * 2016-08-16 2017-01-11 中国烟草总公司广东省公司 Method for comprehensively enhancing tobacco field weeding capacity and disease resisting capacity of tobacco seedlings and reducing malformation
CN108728386A (en) * 2018-06-20 2018-11-02 荆门市拓达科技有限公司 A kind of preparation method of glyphosate solid-state degradation bacterial agent
CN116656376A (en) * 2023-05-31 2023-08-29 辽宁省微生物科学研究院 Preparation method and application of microbial conditioner

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CN104120100A (en) * 2014-07-11 2014-10-29 湖南省农业科学院 Endogenous bacillus megatherium and applications thereof in restoration of quinclorac phytotoxicity

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CN101215536A (en) * 2007-12-29 2008-07-09 浙江工业大学 Method for producing dejection degradation bacterium preparation
CN101724596A (en) * 2010-01-11 2010-06-09 北京联合大学 Method for culturing organophosphorus pesticide degrading bacteria
CN103865853A (en) * 2014-03-17 2014-06-18 中国农业科学院烟草研究所 Efficient degradation bacterium for quinclorac as well as use and use method thereof
CN104120100A (en) * 2014-07-11 2014-10-29 湖南省农业科学院 Endogenous bacillus megatherium and applications thereof in restoration of quinclorac phytotoxicity

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106305178A (en) * 2016-08-16 2017-01-11 中国烟草总公司广东省公司 Method for comprehensively enhancing tobacco field weeding capacity and disease resisting capacity of tobacco seedlings and reducing malformation
CN106305178B (en) * 2016-08-16 2019-12-03 中国烟草总公司广东省公司 A kind of comprehensive method for improving tobacco field weeding and tobacco seedlings disease resistance and reduce teratogenesis generation
CN106148248A (en) * 2016-09-06 2016-11-23 山东农业大学 A kind of Camellia sinensis special organophosphor decomposer YP8 and bacterial preparation process thereof
CN106148248B (en) * 2016-09-06 2018-11-30 山东农业大学 A kind of dedicated organic phosphorus decomposer YP8 of tea tree and its bacterial preparation process
CN108728386A (en) * 2018-06-20 2018-11-02 荆门市拓达科技有限公司 A kind of preparation method of glyphosate solid-state degradation bacterial agent
CN116656376A (en) * 2023-05-31 2023-08-29 辽宁省微生物科学研究院 Preparation method and application of microbial conditioner
CN116656376B (en) * 2023-05-31 2024-01-05 辽宁省微生物科学研究院 Preparation method and application of microbial conditioner

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Application publication date: 20151021