CN101693883B - Microbial agent for degrading phoxim pesticide and preparation method thereof - Google Patents
Microbial agent for degrading phoxim pesticide and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a microbial agent for degrading phoxim pesticide and a preparation method thereof, belonging to the technical field of microbe utilization. In the invention, a pseudomonas YJL-1 strain obtained by selecting is used as a culture which has the preservation number of CGMCC NO.3211; and a microbial agent product is prepared by the steps as follows: (1) preparing a culture medium; (2) preparing the culture; (3) activating the culture; (4) amplifying the culture by shaking a flask; (5) fermenting the microbial agent; and the like. When the microbial agent is used, the microbial agent is diluted by 500-1000 times and applied to soil and crops by spraying, then the microbial agent can degrade the residual phoxim pesticide on the soil and crops, and the degrading efficiency is as high as 80-90%. The microbial agent for degrading phoxim pesticide can be popularized and applied in departments of agricultural production, agricultural product processing, and the like.
Description
Technical field
The present invention relates to technical field of microbial, but relate in particular to a kind of bacterial strain and the microbial inoculum that utilizes this bacterial strain to be prepared from and the preparation method of this microbial inoculum of efficiently degrading organophosphorus pesticides.
Background technology
The present annual agricultural chemicals ultimate production of China has reached 500,000 tons, and up to now, the accumulation consumption of China's agricultural chemicals has reached more than 400 ten thousand tons.Sterilant accounts for more than 70% of all agricultural chemicals, and wherein the organic phosphorous insecticide of high poison accounts for more than 70% of all sterilants again, and promptly the organic phosphorous insecticide of high poison accounts for more than 50% of whole agricultural chemicals market.In 6 insecticide varieties of YO more than ten thousand tons, there are 5 to be organic phosphorous insecticide.See that from present agricultural chemicals service condition the riskiest pesticide usage quantity is big, access times frequently are the major causes that causes some food, particularly vegetables, fruit pesticide residue to exceed standard.For the low agricultural chemicals of toxicity, if consumption is big or improper use can cause the pollution to some food too.
Organophosphorus pesticide (OPs) occupies consequence in sterilant; Be to produce and use maximum pesticide species in the world; The organophosphorus pesticide of China accounts for 70% in sterilant, descended to some extent through adjustment in recent years, but still be the important means of control crop pests and sick entomophila insect.Yet,, become important chemical environment pollutent day by day because the toxicity of organophosphorus pesticide along with being widely used, is also brought severe side effect.Evidence suggests that OPs has mutagenicity and causes birth defect, mammiferous nerve is relevant with OPs mostly with disease of immune system, and these diseases comprise mad cow disease, the complexities of the Gulf War, parkinsonism etc.How to eliminate organophosphorus pesticide to environment and human harm, become problem concerning the human health and the national economic development.
Microbiological deterioration is considered to eliminate a kind of effective measure of organophosphorus pesticide, is the popular direction of Chinese scholars research always.Utilizing biology or biologics to come the biological renovation method of degradation of contaminant to have advantages such as nontoxic, noresidue, non-secondary pollution, is to eliminate and residual a kind of safe, effective, the inexpensive method of detoxifcation high concentration agricultural chemical.But there is poor growth in the mikrobe of existing degrading organic phosphor, lacks competitive edge, problems such as degradation property instability.Bacterium is owing to multiple adaptive faculty on its tool biochemistry and bring out sudden change easily, thereby the strains for degrading ability of in the mikrobe of degrading pesticide, occupying main status, especially Rhodopseudomonas (Pseudomonas sp.) is stronger.
Summary of the invention
The present invention seeks to; To in the present microbiological deterioration technology existing lack growth fast, cultivate present situation simple, can the residual microbial strains of efficiently degrading organophosphorus pesticides; Provide a kind of fast growth, cultural method simple, can be efficient, stablize the bacterial strain of organophosphorus insecticide such as degrading phoxim, and the microbial inoculum of the soil that utilizes this bacterial strain production to be used to administer organophosphorus insecticide to cause, farm crop and products thereof pollution and the preparation method of this microbial inoculum.
The object of the invention is achieved through following technical scheme.
Mikrobe provided by the present invention is Rhodopseudomonas (Pseudomonassp.) bacterial strain of a strain called after YJL-1, and this bacterial strain obtains through following cultivation, separation, screening:
1) we get the active sludge 2g that in Hangzhou insecticide factory treatment tank, gathers in October, 2007, add in the 100ml enrichment medium, and the prescription of this substratum is: NaCl 0.5-1.5g/L, NH
4NO
30.5-1.5g/L, K
2HPO
41.0-2.0g, KH
2PO
40.3-0.8g/L, MgSO
47H
2O 0.1-0.3g/L, yeast extract 0.05-0.2g/L behind the accent pH7.0-7.2, presses 50-100mg/L and adds Volaton, and in 28-37 ℃, the 120-180rpm shaking table is cultivated;
2) the every 1-2 of above-mentioned nutrient solution week switching culture transferring once in the inoculum size access fresh culture with 5-10%, was so tamed 3-6 month; Final culture bacteria liquid is carried out bacterium with plate dilution method separate, it is individual therefrom to obtain single bacterium colony more than 30 that growth is fast, bacterium colony is regular;
3) with after cultivating in single bacterium access LB test tube; Add and contain in the minimal medium of 50-100mg/L Volaton detection Volaton content behind the 3d, the checking of process degradation property; Screening obtains the highest bacterial strain of degradation rate (being numbered X-1); Identify through Physiology and biochemistry, confirm that this bacterial strain belongs to Rhodopseudomonas (Pseudomonas sp.), called after YJL-1; Be stored in the frozen pipe of LB inclined-plane or glycerine, carry out activation on the substratum of Volaton containing during use; The principal character of this bacterial strain is: Gram-negative, nonspore-bearing rod bacterium, bacterium colony are faint yellow, regular edges, and smooth surface, corrugationless, aerobic, changing can be peculiar, oxidase negative, the catalase positive; Optimum growth temperature is 28-30 ℃, and the righttest growth pH is 7.0-7.2, in minimal medium, is sole carbon source growth with (100mg/L) Volaton, and degradation rate reached 90-95% in 3-5 days;
This bacterium is stored in China Committee for Culture Collection of Microorganisms common micro-organisms center on July 22nd, 2009, and deposit number is CGMCC No.3211.
A kind of microbiobacterial agent of degrading phoxim pesticide, this microbial inoculum can LB be inclined-plane or liquid nutrient medium, with NaCl 0.5-1.5g/L, NH
4NO
30.5-1.5g/L, K
2HPO
41.0-2.0g, KH
2PO
40.3-0.8g/L, MgSO
47H
2O 0.1-0.3g/L, yeast extract 0.05-0.2g/L, accent pH7.0-7.2 is a minimal medium; With glucose 5-15g/L, yeast extract paste 1-2g/L, (NH
4)
2SO
41.0-2.0g/L NaCl0.5-1g/L is a fermention medium; Rhodopseudomonas (Pseudomonas sp.) the YJL-1 bacterial strain that with the deposit number is CGMCC No.3211 is a bacterial classification, activated, amplification and bacteria fermentation preparation and obtain.
A kind of preparation method of degrading phoxim pesticide microbiobacterial agent, this method is carried out according to the following steps:
1) various culture medium preparation: comprise
1. LB substratum: 2.5-7.5g/LNaCl, 2.5-7.5g/L yeast extract paste, 5-15g/L peptone;
2. minimal medium: NaCl 0.5-1.5g/L, NH
4NO
30.5-1.5g/L, K
2HPO
41.0-2.0g/L, KH
2PO
40.3-0.8g/L, MgSO
47H
2O 0.1-0.3g/L, yeast extract 0.05-0.2g/L transfers pH7.0-7.2;
3. fermention medium: glucose 5-15g/L, yeast extract paste 1-2g/L, (NH
4)
2SO
41.0-2.0g/L, NaCl0.5-1g/L;
2) preparation of bacterial classification: with deposit number be the YJL-1 inoculation of CGMCC No.3211 on the LB inclined-plane, cultivate 18-24h for 30-35 ℃, the inclined-plane be stored in 0-4 ℃ of refrigerator and cooled hide, subsequent use; Or with the YJL-1 inoculation in the LB test tube, 120-180rpm vibration is cultivated 18-24h for 30-35 ℃, bacterium liquid is added in the glycerine of 50-70%-40 ℃ to-80 ℃ cryopreservation, subsequent use;
3) activation of bacterial classification: from LB inclined-plane or the frozen pipe of glycerine, rule to solid LB flat board 30-35 ℃ of activation culture 18-24h; Single colony inoculation after the activation in the LB liquid nutrient medium, is added the 100mg/L Volaton, and 30-35 ℃, the 120-180rpm shaking table is cultured to logarithmic phase; With above-mentioned medium centrifugal, after pH7.0-7.2 phosphoric acid buffer (PSB) cleaning, contain in the minimal medium of 50-100mg/L Volaton with the adding of 5-10% inoculum size; 30 ℃, the 150rpm shaking table is cultivated, and detects Volaton concentration behind the 3d; When degradation rate reaches 90% when above, use as bacterial classification;
4) shake the amplification of bottle bacterial classification: according to the method for step (3) with yeast culture to logarithmic phase; The inoculum size of pressing 5-10% inserts in the 100mlLB substratum; At pH7.0-7.2, shaking speed 120-150rpm cultivates 18-24h down, and bacterium liquid directly is used for inoculation or 0-4 ℃ of refrigerator and cooled Tibetan, subsequent use;
5) bacteria fermentation: step (1) fermention medium is added in the fermentor tank, and temperature keeps after 30 minutes for 121 ℃ in jar, and pressurize is carried out steam high-temperature sterilization postcooling to 30-35 ℃ between the 0.02-0.18MPa; Under the flame protection, in seeding tank, insert step (4) through the inoculation valve rapidly and shake a bottle bacterial classification, inoculum size is 5-10%; The stirrer rotating speed is 110-220 rev/min, temperature 30-35 ℃, and air flow 2-4m
3/ h, tank pressure 0.02-0.18MPa, pH nature is cultivated and was reached 10 to the thalline number in 18-24 hour
8Individual/more than the ml, the zymocyte liquid can in the aseptic plastic bottle the microbial inoculum product, directly use or under room temperature, preserve.
Microbial inoculum of the present invention dilutes 500-1000 doubly with microbial inoculum when using, spray on soil, crop or agricultural-food, can effectively degrade to the phoxim pesticide residue on the above-mentioned object.
Beneficial effect of the present invention:
1, the pseudomonas YJL-1 bacterial strain that the present invention separates, screening obtains; In the bacterium of the relevant degrading phoxim of having reported at present, still belong to the first time; This strains for degrading phoxim performance is stablized by force; Not only can in triangular flask, degrade; The all right phoxim of degrade residual on soil, crops and agricultural product, degradation efficiency is seen embodiment 4,5,6 up to 80-90%();
2, the YJL-1 bacterial strain has energetic; Fast growth; Advantages such as cultural method is simple, and degradation capability is stable, and adaptive capacity to environment is strong; In the LB substratum, only need to cultivate a 18-24h and just can gather in the crops, be suitable for reparation (seeing embodiment 1,2,3) physical environment such as soil and agricultural-food pesticide residue;
3, this bacterium is expected to reducing the agricultural-food pesticide residue, applied in the pesticide residue in the degraded edatope, environmental pollution improvement with protect people to play a role aspect healthy.
Embodiment
Through following examples the present invention is done more detailed explanation, but following examples only are illustrative, the present invention does not receive the restriction of these embodiment.
Embodiment 1: (degrading phoxim pesticide microbial preparation preparation 1)
Carry out according to the following steps:
1) various culture medium preparation: comprise
1. LB substratum: 2.5g/LNaCl, 7.0g/L yeast extract paste, 5g/L peptone;
2. minimal medium: NaCl 0.5g/L, NH
4NO
31.5g/L, K
2HPO
41.0g/L, KH
2PO
40.8g/L, MgSO
47H
2O 0.2g/L, yeast extract 0.2g/L transfers pH7.0;
3. fermention medium: glucose 10g/L, yeast extract paste 1g/L, (NH
4)
2SO
42.0g/L, NaCl 0.5g/L;
2) preparation of bacterial classification: with deposit number be the YJL-1 inoculation of CGMCC No.3211 on the LB inclined-plane, cultivate 24h for 30 ℃, the inclined-plane be stored in 0 ℃ of refrigerator and cooled hide, subsequent use;
3) activation of bacterial classification: from the LB inclined-plane, rule to solid LB flat board 30 ℃ of activation culture 24h; Single colony inoculation after the activation in the LB liquid nutrient medium, is added the 100mg/L Volaton, and 35 ℃, the 180rpm shaking table is cultured to logarithmic phase; With above-mentioned medium centrifugal, after pH7.0-7.2 phosphoric acid buffer (PSB) cleaning, contain in the minimal medium of 50mg/L Volaton with the adding of 5% inoculum size; 30 ℃, the 150rpm shaking table is cultivated, and detects Volaton concentration behind the 3d; When degradation rate reaches 90% when above, use as bacterial classification;
4) shake the amplification of bottle bacterial classification: according to the method for step (3) with yeast culture to logarithmic phase; Inoculum size by 10% inserts in the 100mlLB substratum; At pH7.0-7.2, shaking speed 150rpm cultivates 18h down, and bacterium liquid directly is used for inoculation or 0-4 ℃ of refrigerator and cooled Tibetan, subsequent use;
5) bacteria fermentation: step (1) fermention medium is added in the fermentor tank, and temperature keeps after 30 minutes for 121 ℃ in jar, and pressurize is carried out steam high-temperature sterilization postcooling to 30-35 ℃ to 0.02MPa; Under the flame protection, in seeding tank, insert step (4) through the inoculation valve rapidly and shake a bottle bacterial classification, inoculum size is 5%; The stirrer rotating speed is 110 rev/mins, 35 ℃ of temperature, air flow 2m
3/ h, tank pressure 0.18MPa, pH nature was cultivated 18 hours, the zymocyte liquid can in the aseptic plastic bottle the microbial inoculum product, directly application or under room temperature, preserve; Detect through enumeration, the thalline number reaches 10 in the cultured bacterium liquid
8Individual/more than the ml.
Embodiment 2: (degrading phoxim pesticide microbial preparation preparation 2)
In this example, the various culture medium preparation of step 1) are: 1. LB substratum: 4.5g/LNaCl, 4.0g/L yeast extract paste, 10g/L peptone; 2. minimal medium: NaCl 1.0g/L, NH
4NO
31.0g/L, K
2HPO
41.5g/L, KH
2PO
40.5g/L, MgSO
47H
2O 0.3g/L, yeast extract 0.05g/L transfers pH7.1; 3. fermention medium: glucose 5g/L, yeast extract paste 1.5g/L, (NH
4)
2SO
41.5g/L, NaCl 0.75g/L;
Step 2) preparation of bacterial classification: in the LB test tube, the 180rpm vibration is cultivated 21h for 30 ℃ with the YJL-1 inoculation, in the glycerine with bacterium liquid adding 60%, and-80 ℃ of cryopreservation, subsequent use;
The activation of step 3) bacterial classification: from the frozen pipe of LB glycerine, rule to solid LB flat board 33 ℃ of activation culture 21h; Single colony inoculation after the activation in the LB liquid nutrient medium, is added the 100mg/L Volaton, and 30 ℃, the 150rpm shaking table is cultured to logarithmic phase; With above-mentioned medium centrifugal, after pH7.0-7.2 phosphoric acid buffer (PSB) cleaning, contain in the minimal medium of 100mg/L Volaton with the adding of 7% inoculum size;
Step 4) is shaken a bottle bacterial classification amplification: the inoculum size by 8% inserts in the 100mlLB substratum, and at pH7.0-7.2, shaking speed 135rpm cultivates 21h down;
The step 5) bacteria fermentation: pressurize is to 0.10MPa in the fermentor tank; Inoculum size is 8%; The stirrer rotating speed is 150 rev/mins, 30 ℃ of temperature, air flow 3m
3/ h, tank pressure 0.02MPa, the pH nature was cultivated 21 hours;
All the other steps, technology are same as embodiment 1.Detect through enumeration, the thalline number reaches 10 in the cultured bacterium liquid
8Individual/more than the ml.
Embodiment 3: (degrading phoxim pesticide microbial preparation preparation 3)
In this example, the various culture medium preparation of step 1) are: 1. LB substratum: 7.5g/LNaCl, 2.5g/L yeast extract paste, 15g/L peptone; 2. minimal medium: NaCl 1.5g/L, NH
4NO
30.5g/L, K
2HPO
42.0g/L, KH
2PO
40.3g/L, MgSO
47H
2O 0.1g/L, yeast extract 0.1g/L transfers pH7.2; 3. fermention medium: glucose 15g/L, yeast extract paste 2.0g/L, (NH
4)
2SO
41.0g/L, NaCl 1.0g/L;
Step 2) preparation of bacterial classification: the YJL-1 inoculation on the LB inclined-plane, is cultivated 18h for 35 ℃, and the inclined-plane is stored in 4 ℃ of refrigerator and cooled Tibetan, subsequent use;
The activation of step 3) bacterial classification: from the LB inclined-plane, rule to solid LB flat board 35 ℃ of activation culture 18h; Single colony inoculation after the activation in the LB liquid nutrient medium, is added the 100mg/L Volaton, and 32 ℃, the 120rpm shaking table is cultured to logarithmic phase; With above-mentioned medium centrifugal, after pH7.0-7.2 phosphoric acid buffer (PSB) cleaning, contain in the minimal medium of 75mg/L Volaton with the adding of 10% inoculum size;
Step 4) is shaken a bottle bacterial classification amplification: the inoculum size by 5% inserts in the 100mlLB substratum, and at pH7.0-7.2, shaking speed 120rpm cultivates 24h down;
The step 5) bacteria fermentation: pressurize is to 0.18MPa in the fermentor tank; Inoculum size is 10%; The stirrer rotating speed is 220 rev/mins, 33 ℃ of temperature, air flow 4m
3/ h, tank pressure 0.10MPa, the pH nature was cultivated 24 hours;
All the other steps, technology are same as embodiment 1.Detect through enumeration, the thalline number reaches 10 in the cultured bacterium liquid
8Individual/more than the ml.
Embodiment 4: (organophosphorus degrading bacteria of the present invention is to the degradation effect test of Volaton)
YJL-1 bacterial strain list colony inoculation after the activation in the LB liquid nutrient medium, is added the 100mg/L Volaton, and 30 ℃, the 150rpm shaking table is cultivated 24h; With above-mentioned medium centrifugal, with phosphoric acid buffer (pH7.0-7.2) clean, centrifugal 2 times, again with phosphoric acid buffer with thalline OD
600Transfer to 0.1-0.2, contain in the minimal medium of 100mg/L Volaton with the adding of 10% inoculum size, 28-30 ℃, the 120-150rpm shaking table is cultivated, and detects Volaton concentration and OD behind the 3d
600Value, checking degradation effect and thalli growth situation.Through detecting, the Volaton degradation rate is more than 90%, thalline OD
600Reach more than 0.6, explain that the YJL-1 bacterial strain has good degradation effect to Volaton, and can utilize the Volaton growth.Described LB liquid nutrient medium and minimal medium are same as embodiment 1,2 or 3.
Embodiment 5: (degraded that organic phosphorus degrading microbial inoculum of the present invention is residual to Volaton on the green vegetables)
The test place is test base, academy of agricultural sciences, Zhejiang Province, and the time is in October, 2008, and each tests the sub-district area is 1 mu, contrast and each 3 of processing sub-districts.On the green vegetables blade face of contrast and processing, all spray the Volaton solution of 0.5mg/mL, on the green vegetables blade face of handling, spray the YJL-1 microbial inoculum diluent (about 10 of embodiment 1 preparation behind the 3d again
8/ mL), 6kg/ mu.To spray back the 3rd, 7,15, measure the content (detecting) of Volaton in the green vegetables during 21d respectively by agricultural product quality supervision and inspection center of the Ministry of Agriculture (Hangzhou).Result's (table 1) shows that the organophosphorus comparison has dropped to about 50% of contrast according to descending to some extent at 6d in the 2d green vegetables after using the YJL-1 microbial inoculum; And during to 14d; Compare with contrast, handle degraded about 70%, during to 20d; The concentration of Volaton has dropped to below the 0.01mg/kg in the processing, almost completely degraded.
Table 1 YJL-1 is to the degraded (Volaton residual quantity mg/kg) of Volaton on the green vegetables
Embodiment 6: (degraded that organic phosphorus degrading microbial inoculum of the present invention is residual to Volaton in the soil)
Soil is taken from experiment field, academy of agricultural sciences, Zhejiang Province, and soil type is a yellowish soil, organic content 14.5g/kg, nitrogenous 1.2g/kg, pH value 7.2, soil moisture content 60-70%.Select the veneer of soil of no phoxim pesticide residue in the soil, each sample 10kg soil, the artificial Volaton concentration of adding is to 50mg/kg in this soil again, and the YJL-1 bacterium liquid usage quantity of embodiment 1 preparation is 10
8/ g dry ground.After executing bacterium, respectively at the 7th, 14,20d measures the content of Volaton in the soil.The result shows that in the natural soils of inoculation YJL-1, the degraded of Volaton will be compared according to much fast in the soil, has accelerated the degraded of agricultural chemicals significantly.After adding bacterium 7d, Volaton has only degraded 24.4% in the control soil, and Volaton has degraded 71% in the soil of inoculation YJL-1.To 20d, Volaton concentration has dropped to 5mg/kg in the soil of inoculation YJL-1, and the concentration of Volaton still has 24.9mg/kg in control soil.Obviously, YJL-1 has quickened the degraded of Volaton in natural soils.
Claims (3)
1. Rhodopseudomonas Pseudomonas sp.YJL-1 bacterial strain, its bacterial strain preserving number is CGMCC NO.3211.
2. the microbiobacterial agent of a degrading phoxim pesticide is characterized in that this microbial inoculum is is inclined-plane or liquid nutrient medium with LB; With NaCl 0.5-1.5g/L, NH
4NO
30.5-1.5g/L, K
2HPO
41.0-2.0g, KH
2PO
40.3-0.8g/L, MgSO
47H
2O 0.1-0.3g/L, yeast extract 0.05-0.2g/L, accent pH7.0-7.2 is a minimal medium; With glucose 5-15g/L, yeast extract paste 1-2g/L, (NH
4)
2SO
41.0-2.0g/L NaCl0.5-1g/L is a fermention medium; The Rhodopseudomonas Pseudomonas sp.YJL-1 bacterial strain that with the deposit number is CGMCC No.3211 is a bacterial classification, activated, amplification and bacteria fermentation preparation and obtain.
3. the preparation method of the microbiobacterial agent of a degrading phoxim pesticide is characterized in that carrying out according to the following steps:
1) various culture medium preparation: comprise
1. LB substratum: 2.5-7.5g/LNaCl, 2.5-7.5g/L yeast extract paste, 5-15g/L peptone;
2. minimal medium: NaCl 0.5-1.5g/L, NH
4NO
30.5-1.5g/L, K
2HPO
41.0-2.0g/L, KH
2PO
40.3-0.8g/L, MgSO
47H
2O 0.1-0.3g/L, yeast extract 0.05-0.2g/L transfers pH7.0-7.2;
3. fermention medium: glucose 5-15g/L, yeast extract paste 1-2g/L, (NH
4)
2SO
41.0-2.0g/L, NaCl0.5-1g/L;
2) preparation of bacterial classification: with deposit number be the YJL-1 inoculation of CGMCC No.3211 on the LB inclined-plane, cultivate 18-24h for 30-35 ℃, the inclined-plane be stored in 0-4 ℃ of refrigerator and cooled hide, subsequent use; Or with the YJL-1 inoculation in the LB test tube, 120-180rpm vibration is cultivated 18-24h for 30-35 ℃, bacterium liquid is added in the glycerine of 50-70%-40 ℃ to-80 ℃ cryopreservation, subsequent use;
3) activation of bacterial classification: from LB inclined-plane or the frozen pipe of glycerine, rule to solid LB flat board 30-35 ℃ of activation culture 18-24h; Single colony inoculation after the activation in the LB liquid nutrient medium, is added the 100mg/L Volaton, and 30-35 ℃, the 120-180rpm shaking table is cultured to logarithmic phase; With above-mentioned medium centrifugal, after the cleaning of pH7.0-7.2 phosphoric acid buffer, contain in the minimal medium of 50-100mg/L Volaton with the adding of 5-10% inoculum size; 30 ℃, the 150rpm shaking table is cultivated, and detects Volaton concentration behind the 3d; When degradation rate reaches 90% when above, use as bacterial classification;
4) shake the amplification of bottle bacterial classification: according to the method for step (3) with yeast culture to logarithmic phase; The inoculum size of pressing 5-10% inserts in the 100mlLB substratum; At pH7.0-7.2, shaking speed 120-150rpm cultivates 18-24h down, and bacterium liquid directly is used for inoculation or 0-4 ℃ of refrigerator and cooled Tibetan, subsequent use;
5) bacteria fermentation: step (1) fermention medium is added in the fermentor tank, and temperature keeps after 30 minutes for 121 ℃ in jar, and pressurize is carried out steam high-temperature sterilization postcooling to 30-35 ℃ between the 0.02-0.18MPa; Under the flame protection, in seeding tank, insert step (4) through the inoculation valve rapidly and shake a bottle bacterial classification, inoculum size is 5-10%, and the stirrer rotating speed is 110-220 rev/min, temperature 30-35 ℃, and air flow 2-4m
3/ h, tank pressure 0.02-0.18MPa, pH nature is cultivated and was reached 10 to the thalline number in 18-24 hour
8Individual/more than the ml, the zymocyte liquid can in the aseptic plastic bottle the microbial inoculum product, directly use or under room temperature, preserve.
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| CN107716530A (en) * | 2017-09-25 | 2018-02-23 | 南京索益盟检测技术有限公司 | A kind of soil-repairing agent of absorption degradation organic agricultural chemicals and preparation method thereof |
| CN109679875B (en) * | 2019-01-17 | 2020-06-23 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Delftia sp and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1800353A (en) * | 2005-12-23 | 2006-07-12 | 南京农业大学 | Bacterium for degrading phoxim pesticide residue and produced bacterium formulation |
| CN101139559A (en) * | 2006-09-05 | 2008-03-12 | 许雷 | Novel bacterial strain for highly effective degradation of chrysanthemum ester and organophosphorus pesticide and uses thereof |
-
2009
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1800353A (en) * | 2005-12-23 | 2006-07-12 | 南京农业大学 | Bacterium for degrading phoxim pesticide residue and produced bacterium formulation |
| CN101139559A (en) * | 2006-09-05 | 2008-03-12 | 许雷 | Novel bacterial strain for highly effective degradation of chrysanthemum ester and organophosphorus pesticide and uses thereof |
Non-Patent Citations (2)
| Title |
|---|
| 张瑞福等.辛硫磷降解菌X21 的分离鉴定及降解性状的初步研究.《环境科学学报》.2003,第23卷(第3期),全文. * |
| 沈雨佳等.辛硫磷降解菌XSP21 的分离、鉴定及其降解特性研究.《环境科学》.2007,第28卷(第12期),全文. * |
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