CN104126614A - Preparation method for biological pesticide - Google Patents
Preparation method for biological pesticide Download PDFInfo
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- CN104126614A CN104126614A CN201410403112.1A CN201410403112A CN104126614A CN 104126614 A CN104126614 A CN 104126614A CN 201410403112 A CN201410403112 A CN 201410403112A CN 104126614 A CN104126614 A CN 104126614A
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Abstract
The invention belongs to a preparation method for a biological pesticide. The strain of the biological pesticide is called stem borers-killing bacillus. The preparation method for the biological pesticide comprises the steps of preparing a mother culture medium, performing inoculation and rejuvenation, expanding production seeds, making and detecting first-class solid seeds and the like. The pesticide does not have any toxic and side effects on a human ecological circle system and crops. The method is simple and practical; the pesticide is easy to prepare and produce; the preparation and use costs are low; the pesticide has the advantages of favorability for transportation and storage and good insect killing effect.
Description
Technical field
The invention belongs to a kind of preparation method of biopesticide.
Background technology
Agricultural chemicals energy kill pests improves crop yield, but it also can remain on crops or enter in the human ecology circulatory system, harm humans healthy, the pollution channel of crops is by the direct absorption of its base of leaf with from root osmotic absorption, discharging of chemical pesticide, as the residue organochlorine of agricultural chemicals, organic phosphor etc., the vegetables of the residue that people are contained agricultural chemicals in picked-up every day, fruit, drinking-water all can endanger physical and mental health.
But the biopesticide of controlling worm with worm is done without pair, can be crop growth better, provide safeguard, and without any side effects to the human ecology circulatory system and crops.But biopesticide is of less types at present, preparation method is complicated, and cost is high, is difficult to universal use.
Summary of the invention
The preparation method who the object of this invention is to provide a kind of biopesticide, this agricultural chemicals is without any side effects to the human ecology circulatory system and crops, and the method is simple and practical, is easy to make produce, making and use cost are low, are beneficial to the advantage of transportation preservation and good disinsection effect.
For this reason, the name of this biopesticide bacterial classification of the present invention is called bacillus thuringensis, and this biological preparation method comprises the steps:
(1) prepare mother culture media, get without muscle, without oily beef 500g chopping, stir into meat slurry, add water 1000ml, soak 1 hour, boil, be incubated 15～20 minutes, stop insulation, treat that meat soup temperature is down to 30 ℃, add Egg-white 15g, stir, filter impurity elimination, add water and supply filtrate to 1000ml, then in filtrate, add successively glucose 1g, sodium chloride 4～6g, potassium dihydrogen phosphate 1.3g, dipotassium hydrogen phosphate 1.5g, stirs, then adds agar 15g, be stirred to agar and dissolve in filtrate, with sodium chloride, adjusting the pH value of filtrate is 7.2;
(2) dress test tube, every test tube injects filtrate 4ml, and the sterilization 30 minutes at 121 ℃ of test tube and filtrate, takes out test tube, is put into inclined-plane, tilting 45 ° of test tube;
(3) inoculation, gets without infecting a bacterial classification of 6～8 test tube accesses of test tube;
(4) cultivate, the test tube of access bacterium, at 25 ℃～30 ℃, is cultivated to 5-7 days, can obtain first female kind;
(5) rejuvenation, by first female kind theing obtain, repeating step (3) to (4) carries out rejuvenation of spawn 1～2 time;
(6) making of one-level solid seed, the medium of its one-level solid seed is comprised of wheat bran 65g, beancake powder 25g, steamed beef soup 10g, peptone 5g, glucose 1g, sodium chloride 2～4g, potassium dihydrogen phosphate 1.4g, dipotassium hydrogen phosphate 1.5g, yeast extract 1.5g, water 1000ml, above-mentioned raw materials is mixed, and adjusting PH is 7.2;
This medium is packed in triangular flask, every bottle of 50g, bottleneck jam-pack cotton plug, its outside is wrapped one deck brown paper again;
This bottle is put to 121 ℃ of sterilizings 30 minutes, naturally cool to 18 ℃～21 ℃;
Every inclined-plane can connect 10 bottles of female bacterial classifications, can a waddy one centimeter of insert eye from material is dark before inoculation, during inoculation, bacterial classification is put into hole, and be then placed in 25-28 ℃ of incubator and cultivate 5 days, obtain one-level solid seed;
(7) expanding production kind, its raw material weight percentage of producing the medium of planting is wheat bran 25%, beancake powder 25%, thin chaff 15%, chaff 35%, fused calcium magnesium phosphate 1%, above-mentioned raw materials is mixed, adding thin rice gruel water to make the weight ratio of itself and above-mentioned raw materials is 1.3: 1, mix, with sodium carbonate, regulating the PH of above-mentioned medium is 9 again;
Pack above-mentioned medium into food steamer, with the sterilizing 40 minutes at 121 ℃ of Large-scale High-Pressure autoclave, be cooled to 28 ℃～32 ℃, by putting one bottle, one-level solid seed in every 100 jin of these medium, be mixed, then be layered on the bamboo bed that can ventilate through underlay one deck white nylon cloth of sterilizing, this medium thickness is 22～26 centimeters, cap rock gauze above, stir every day once, in the culturing room of 28 ℃, cultivate 3-5 days, on it, all cover with after mycelia, dry or 60 ℃ of low temperature dryings, must produce and plant;
(8) detecting, take 1g and produce and kind add aqua sterilisa 100ml, after dilution, under 600 power microscopes and at hemocytometer, on several pieces, detect, measure the bacteria containing amount of finished product for reaching the standard of hundred million of 50-100, is the dry spice of desinsection;
(9) using method;
(1), the dry spice of 500g desinsection can be watered 25-30kg, spraying is executed and is killed, loose painting;
(2), the dry spice of 500g desinsection adds 5kg warm water, soaks filtration in 30 minutes, filtrate adds water 250-300kg, spraying is executed and is killed, loose painting;
While using this biopesticide, should be than the early 2-3 days that executes of other agricultural chemicals, temperature is used above at 20 ℃;
Kill object, the snout moth's larva of paddy rice class, rice plant skipper, rice fulgorid, tortrix moth, leafhopper, rice borer bell, leaf springtail, thrips; The cabbage caterpillar of greengrocery, diamond-back moth, bridging worm, mythimna separata, cutworm; The slug moth of fruit tree kind, skinworm, urge cotton worm, scale insect, two spotted spider mite, longicorn, tea caterpillar; The pine caterpillars of forestry class, phosphorus wing larva; The moths attracted by lamplight, Limonene fiber crops worm of cotton class; The duckweed snout moth's larva of duckweed class, duckweed ash snout moth's larva.
The described mother culture media of preparing, get without muscle, without oily beef 500g chopping, stir into meat slurry, add water 1000ml, soak 1 hour, boil, be incubated 15～20 minutes, stop insulation, treat that meat soup temperature is down to 30 ℃, add Egg-white 15g, stir, filter impurity elimination, add water and supply filtrate to 1000ml, then in filtrate, add successively glucose 1g, sodium chloride 4～6g, potassium dihydrogen phosphate 1.3g, dipotassium hydrogen phosphate 1.5g, stirs, then adds agar 15g, be stirred to agar and dissolve in filtrate, with sodium chloride, adjusting the pH value of filtrate is 7.2.
The medium of its described one-level solid seed is comprised of wheat bran 65g, beancake powder 25g, steamed beef soup 10g, peptone 5g, glucose 1g, sodium chloride 2～4g, potassium dihydrogen phosphate 1.4g, dipotassium hydrogen phosphate 1.5g, yeast extract 1.5g, water 1000ml, above-mentioned raw materials is mixed, and adjusting PH is 7.2.
Described rejuvenation of spawn can be undertaken by other traditional method.
The bacteria containing amount of the described dry spice of desinsection is the standard of hundred million of 50-100.
Said method has reached object of the present invention.
Agricultural chemicals of the present invention is without any side effects to the human ecology circulatory system and crops, and the method is simple and practical, is easy to make produce, and making and use cost are low, is beneficial to the advantage of transportation preservation and good disinsection effect.
Test shows that killing rate of the present invention is more than 85%, has good result of use.Making and use cost are only 50% left and right of traditional pesticide cost, and production environment does not require, and are applicable to rural area individual making, can significantly reduce desinsection expense.
Embodiment
A kind of preparation method of biopesticide:
The name of this biopesticide bacterial classification is called bacillus thuringensis, and this biological preparation method comprises the steps:
(1) prepare mother culture media, get without muscle, without oily beef 500g chopping, stir into meat slurry, add water 1000ml, soak 1 hour, boil, be incubated 15～20 minutes, stop insulation, treat that meat soup temperature is down to 30 ℃, add Egg-white 15g, stir, filter impurity elimination, add water and supply filtrate to 1000ml, then in filtrate, add successively glucose 1g, sodium chloride 4～6g, potassium dihydrogen phosphate 1.3g, dipotassium hydrogen phosphate 1.5g, stirs, then adds agar 15g, be stirred to agar and dissolve in filtrate, with sodium chloride, adjusting the pH value of filtrate is 7.2.
(2) dress test tube, every test tube injects filtrate 4ml, and the sterilization 30 minutes at 121 ℃ of test tube and filtrate, takes out test tube, is put into inclined-plane, tilting 45 ° of test tube.
(3) inoculation, gets without infecting a bacterial classification of 6～8 test tube accesses of test tube.
(4) cultivate, the test tube of access bacterium, at 25 ℃～30 ℃, is cultivated to 5-7 days, can obtain first female kind.
(5) rejuvenation, by first female kind theing obtain, repeating step (3) to (4) carries out rejuvenation of spawn 1～2 time.
(6) making of one-level solid seed, the medium of its one-level solid seed is comprised of wheat bran 65g, beancake powder 25g, steamed beef soup 10g, peptone 5g, glucose 1g, sodium chloride 2～4g, potassium dihydrogen phosphate 1.4g, dipotassium hydrogen phosphate 1.5g, yeast extract 1.5g, water 1000ml.Above-mentioned raw materials is mixed, and adjusting PH is 7.2 (adjusting with sodium chloride).
This medium is packed in triangular flask, every bottle of 50g, bottleneck jam-pack cotton plug, its outside is wrapped one deck brown paper again.
This bottle is put to 121 ℃ of sterilizings 30 minutes, naturally cool to 18 ℃～21 ℃.
Every inclined-plane can connect 10 bottles of female bacterial classifications, can a waddy one centimeter of insert eye from material is dark before inoculation, during inoculation, bacterial classification is put into hole, and be then placed in 25-28 ℃ of incubator and cultivate 5 days, obtain one-level solid seed.
(7) expanding production kind, its raw material weight percentage of producing the medium of planting is wheat bran 25%, beancake powder 25%, thin chaff 15%, chaff 35%, fused calcium magnesium phosphate 1%, above-mentioned raw materials is mixed, adding thin rice gruel water to make the weight ratio of itself and above-mentioned raw materials is 1.3: 1, mix, with sodium carbonate, regulating the PH of above-mentioned medium is 9 again.Can via hole intraocular footpath be 1mm sieve or sieve be thin chaff, otherwise be chaff.
Pack above-mentioned medium into food steamer, with the sterilizing 40 minutes at 121 ℃ of Large-scale High-Pressure autoclave, be cooled to 28 ℃～32 ℃, by putting one bottle, one-level solid seed in every 100 jin of these medium, be mixed, then be layered on the bamboo bed that can ventilate through underlay one deck white nylon cloth of sterilizing, this medium thickness is 22～26 centimeters, cap rock gauze above, stir every day once, in the culturing room of 28 ℃, cultivate 3-5 days, on it, all cover with after mycelia, dry or 60 ℃ of low temperature dryings, must produce and plant.
(8) detecting, take 1g and produce and kind add aqua sterilisa 100ml, after dilution, under 600 power microscopes and at hemocytometer, on several pieces, detect, measure the bacteria containing amount of finished product (produce kind) for reaching the standard of hundred million of 50-100, is the dry spice of desinsection.
(9) using method;
(1), the dry spice of 500g desinsection can be watered 25-30kg, spraying is executed and is killed, loose painting;
(2), the dry spice of 500g desinsection adds 5kg warm water, soaks filtration in 30 minutes, filtrate adds water 250-300kg, spraying is executed and is killed, loose painting;
While using this biopesticide, should be than the early 2-3 days that executes of other agricultural chemicals, temperature is used above at 20 ℃;
Kill object, kill object, the snout moth's larva of paddy rice class, rice plant skipper, rice fulgorid, tortrix moth, leafhopper, rice borer bell, leaf springtail, thrips; The cabbage caterpillar of greengrocery, diamond-back moth, bridging worm, mythimna separata, cutworm; The slug moth of fruit tree kind, skinworm, urge cotton worm, scale insect, two spotted spider mite, longicorn, tea caterpillar; The pine caterpillars of forestry class, phosphorus wing larva; The moths attracted by lamplight, Limonene fiber crops worm of cotton class; The duckweed snout moth's larva of duckweed class, duckweed ash snout moth's larva.
The described mother culture media of preparing, get without muscle, without oily beef 500g chopping, stir into meat slurry, add water 1000ml, soak 1 hour, boil, be incubated 15～20 minutes, stop insulation, treat that meat soup temperature is down to 30 ℃, add Egg-white 15g, stir, filter impurity elimination, add water and supply filtrate to 1000ml, then in filtrate, add successively glucose 1g, sodium chloride 4～6g, potassium dihydrogen phosphate 1.3g, dipotassium hydrogen phosphate 1.5g, stirs, then adds agar 15g, be stirred to agar and dissolve in filtrate, with sodium chloride, adjusting the pH value of filtrate is 7.2.
The medium of its described one-level solid seed is comprised of wheat bran 65g, beancake powder 25g, steamed beef soup 10g, peptone 5g, glucose 1g, sodium chloride 2～4g, potassium dihydrogen phosphate 1.4g, dipotassium hydrogen phosphate 1.5g, yeast extract 1.5g, water 1000ml, above-mentioned raw materials is mixed, and adjusting PH is 7.2.
Described rejuvenation of spawn can be undertaken by other traditional method.As transplant activation, separation is selected excellent, improves Media Components and improves optional in meteorological environment.
The bacteria containing amount of the described dry spice of desinsection is the standard of hundred million of 50-100.
Agricultural chemicals of the present invention is without any side effects to the human ecology circulatory system and crops, and the method is simple and practical, is easy to make produce, and making and use cost are low, is beneficial to the advantage of transportation preservation and good disinsection effect.
Claims (5)
1. a preparation method for biopesticide, is characterized in that: the name of this biopesticide bacterial classification is called bacillus thuringensis, and this biological preparation method comprises the steps,
(1) prepare mother culture media, get without muscle, without oily beef 500g chopping, stir into meat slurry, add water 1000ml, soak 1 hour, boil, be incubated 15～20 minutes, stop insulation, treat that meat soup temperature is down to 30 ℃, add Egg-white 15g, stir, filter impurity elimination, add water and supply filtrate to 1000ml, then in filtrate, add successively glucose 1g, sodium chloride 4～6g, potassium dihydrogen phosphate 1.3g, dipotassium hydrogen phosphate 1.5g, stirs, then adds agar 15g, be stirred to agar and dissolve in filtrate, with sodium chloride, adjusting the pH value of filtrate is 7.2;
(2) dress test tube, every test tube injects filtrate 4ml, and the sterilization 30 minutes at 121 ℃ of test tube and filtrate, takes out test tube, is put into inclined-plane, tilting 45 ° of test tube;
(3) inoculation, gets without infecting a bacterial classification of 6～8 test tube accesses of test tube;
(4) cultivate, the test tube of access bacterium, at 25 ℃～30 ℃, is cultivated to 5-7 days, can obtain first female kind;
(5) rejuvenation, by first female kind theing obtain, repeating step (3) to (4) carries out rejuvenation of spawn 1～2 time;
(6) making of one-level solid seed, the medium of its one-level solid seed is comprised of wheat bran 65g, beancake powder 25g, steamed beef soup 10g, peptone 5g, glucose 1g, sodium chloride 2～4g, potassium dihydrogen phosphate 1.4g, dipotassium hydrogen phosphate 1.5g, yeast extract 1.5g, water 1000ml, above-mentioned raw materials is mixed, and adjusting PH is 7.2;
This medium is packed in triangular flask, every bottle of 50g, bottleneck jam-pack cotton plug, its outside is wrapped one deck brown paper again;
This bottle is put to 121 ℃ of sterilizings 30 minutes, naturally cool to 18 ℃～21 ℃;
Every inclined-plane can connect 10 bottles of female bacterial classifications, can a waddy one centimeter of insert eye from material is dark before inoculation, during inoculation, bacterial classification is put into hole, and be then placed in 25-28 ℃ of incubator and cultivate 5 days, obtain one-level solid seed;
(7) expanding production kind, its raw material weight percentage of producing the medium of planting is wheat bran 25%, beancake powder 25%, thin chaff 15%, chaff 35%, fused calcium magnesium phosphate 1%, above-mentioned raw materials is mixed, adding thin rice gruel water to make the weight ratio of itself and above-mentioned raw materials is 1.3: 1, mix, with sodium carbonate, regulating the PH of above-mentioned medium is 9 again;
Pack above-mentioned medium into food steamer, with the sterilizing 40 minutes at 121 ℃ of Large-scale High-Pressure autoclave, be cooled to 28 ℃～32 ℃, by putting one bottle, one-level solid seed in every 100 jin of these medium, be mixed, then be layered on the bamboo bed that can ventilate through underlay one deck white nylon cloth of sterilizing, this medium thickness is 22～26 centimeters, cap rock gauze above, stir every day once, in the culturing room of 28 ℃, cultivate 3-5 days, on it, all cover with after mycelia, dry or 60 ℃ of low temperature dryings, must produce and plant;
(8) detecting, take 1g and produce and kind add aqua sterilisa 100ml, after dilution, under 600 power microscopes and at hemocytometer, on several pieces, detect, measure the bacteria containing amount of finished product for reaching the standard of hundred million of 50-100, is the dry spice of desinsection;
(9) using method;
(1), the dry spice of 500g desinsection can be watered 25-30kg, spraying is executed and is killed, loose painting;
(2), the dry spice of 500g desinsection adds 5kg warm water, soaks filtration in 30 minutes, filtrate adds water 250-300kg, spraying is executed and is killed, loose painting;
While using this biopesticide, should be than the early 2-3 days that executes of other agricultural chemicals, temperature is used above at 20 ℃;
Kill object, the snout moth's larva of paddy rice class, rice plant skipper, rice fulgorid, tortrix moth, leafhopper, rice borer bell, leaf springtail, thrips; The cabbage caterpillar of greengrocery, diamond-back moth, bridging worm, mythimna separata, cutworm; The slug moth of fruit tree kind, skinworm, urge cotton worm, scale insect, two spotted spider mite, longicorn, tea caterpillar; The pine caterpillars of forestry class, phosphorus wing larva; The moths attracted by lamplight, Limonene fiber crops worm of cotton class; The duckweed snout moth's larva of duckweed class, duckweed ash snout moth's larva.
2. by preparation method claimed in claim 1, it is characterized in that: the described mother culture media of preparing, get without muscle, without oily beef 500g chopping, stir into meat slurry, add water 1000ml, soak 1 hour, boil, be incubated 15～20 minutes, stop insulation, treat that meat soup temperature is down to 30 ℃, add Egg-white 15g, stir, filter impurity elimination, add water and supply filtrate to 1000ml, then in filtrate, add successively glucose 1g, sodium chloride 4～6g, potassium dihydrogen phosphate 1.3g, dipotassium hydrogen phosphate 1.5g, stir, add again agar 15g, being stirred to agar dissolves in filtrate, with sodium chloride, adjusting the pH value of filtrate is 7.2.
3. by preparation method claimed in claim 1, it is characterized in that: the medium of its described one-level solid seed is comprised of wheat bran 65g, beancake powder 25g, steamed beef soup 10g, peptone 5g, glucose 1g, sodium chloride 2～4g, potassium dihydrogen phosphate 1.4g, dipotassium hydrogen phosphate 1.5g, yeast extract 1.5g, water 1000ml, above-mentioned raw materials is mixed, and adjusting PH is 7.2.
4. by preparation method claimed in claim 1, it is characterized in that: described rejuvenation of spawn can be undertaken by other traditional method.
5. by preparation method claimed in claim 1, it is characterized in that: the bacteria containing amount of the described dry spice of desinsection is the standard of hundred million of 50-100.
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| CN201410403112.1A CN104126614A (en) | 2014-08-18 | 2014-08-18 | Preparation method for biological pesticide |
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| CN201410403112.1A CN104126614A (en) | 2014-08-18 | 2014-08-18 | Preparation method for biological pesticide |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106035239A (en) * | 2016-05-30 | 2016-10-26 | 中国农业大学 | Method for studying influences exerted by approach-avoidance substances on oviposition behaviors of diamondback moths |
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| CN1374399A (en) * | 2002-04-11 | 2002-10-16 | 中国科学院武汉病毒研究所 | Waxy bacillus and its prepn |
| CN1378784A (en) * | 2002-04-11 | 2002-11-13 | 中国科学院武汉病毒研究所 | Process for preparing bio-insecticide |
| CN1417325A (en) * | 2002-12-14 | 2003-05-14 | 山西省农业科学院植物保护研究所 | Waxy bacillus strain and microbial bactericide producing process with the strain |
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2014
- 2014-08-18 CN CN201410403112.1A patent/CN104126614A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1374399A (en) * | 2002-04-11 | 2002-10-16 | 中国科学院武汉病毒研究所 | Waxy bacillus and its prepn |
| CN1378784A (en) * | 2002-04-11 | 2002-11-13 | 中国科学院武汉病毒研究所 | Process for preparing bio-insecticide |
| CN1417325A (en) * | 2002-12-14 | 2003-05-14 | 山西省农业科学院植物保护研究所 | Waxy bacillus strain and microbial bactericide producing process with the strain |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106035239A (en) * | 2016-05-30 | 2016-10-26 | 中国农业大学 | Method for studying influences exerted by approach-avoidance substances on oviposition behaviors of diamondback moths |
| CN106035239B (en) * | 2016-05-30 | 2018-11-09 | 中国农业大学 | A method of research walks quickly and keeps away substance to Oviposition of Diamondback Moth behavioral implications |
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Application publication date: 20141105 |