CN101658187B - Compound insecticide of bacillus thuringiensis, verticillium lecanii and aconite alkaloid and preparation method thereof - Google Patents
Compound insecticide of bacillus thuringiensis, verticillium lecanii and aconite alkaloid and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a compound insecticide of bacillus thuringiensis, verticillium lecanii and aconite alkaloid and a preparation method thereof. The compound biotic insecticide is mainly mixed through the fermentation liquor of bacillus thuringiensis and the meristem of the verticillium lecanii or blastospore and the aconite alkaloid with proper auxiliary agent according to certain proportion, wherein the mass percentage of the main medicine and the auxiliary agent respectively accounts for 50 percent to 90 percent and 10 percent to 50 percent. The compound insecticide has the advantages that the compound insecticide is broad-spectrum, has control effect to various crop pests, and particularly has obvious effect to homoptera pets, lepidoptera pets and coleopteran pets; the desinsection speed is high; the lasting period is long; pets are difficult to develop drug resistance; the compound insecticide is safe to people and animals, environment and natural enemies; the manufacturing process is simple, the material sources are wide, and the cost is low; and the preparations are multiple, and the application field is wide.
Description
Technical field
The invention belongs to field of biological pesticide, a kind of specifically bacillus thuringiensis, Verticillium lecanii and aconite alkaloid compounded pesticides.
Background technology
In order to bring into play the advantage separately of biopesticide composition, overcome its drawback, reduce and produce agricultural cost, improve drug effect, prolong the lasting period, widen insecticidal spectrum, develop non-toxic, environmental friendly biopesticide to people and animals, environment and natural enemies security, people can carry out reasonably combination collocation to some by compatible biopesticide composition usually, make various complex biological preparations, and various active substances can be augmented mutually.
Now existing experiment products, as: bacillus thuringiensis (Bt) and Avermectin (Av), bacillus thuringiensis, Avermectin and matrine, bacillus thuringiensis, white muscardine fungi and nimbin, matrine and nicotine recompounding biopesticide, but all exist complex manufacturing or active ingredient is indeterminate, the production cost height, a large amount of uses of organic solvent in the plant resource composition leaching process, its safety is reduced, formulation is single, and the undesirable many shortcomings of control efficiency make its Application and Development be restricted.
Summary of the invention
The object of the invention is to provide bacillus thuringiensis, Verticillium lecanii and the aconite alkaloid compounded pesticides of a kind of efficient, environmental protection, safety.
Another object of the present invention is to provide the preparation method of compounded pesticides.
Bacillus thuringiensis of the present invention, Verticillium lecanii and aconite alkaloid compounded pesticides are with main ingredient and additive mixture, and the percentage by weight of main ingredient is that the percentage by weight of 50%-94%, auxiliary agent is 6-50%; Main ingredient is that weight ratio is the mixture of 1: 1: 0.1 bacillus thuringiensis (Bacillus thuringiensis) fermentation product, Verticillium lecanii (Verticillium lecanii) fermentation product and aconite alkaloid.
Main bacteria seed and source: bacillus thuringiensis is that CGMCC1.767 and CGMCC1.1012 were by 1: 1 mixture; Verticillium lecanii CGMCC3.4505 is all available from Chinese microorganism strain preservation center.
Aconite alkaloid is hammered ethanol extract into shape from iron staff in the main ingredient, the preparation method of iron staff hammer ethanol extract is, the iron staff hammer root of drying is ground into 60 order fine powders, with 95% ethanol serves as to extract solvent, the volume of ethanol is three times of iron staff hammer fine powder volume, place altogether and shake bottle jolting extraction 120 hours, filter, filter residue extracted 120 hours with the ethanol solvent jolting of the long-pending amount of triploid again, filter, filter residue merges all filtrates with solvent washing 3 times, decompression recycling ethanol promptly gets the iron staff hammer ethanol extract that contains total aconite alkaloid 82.3%.
The preparation method of bacillus thuringiensis of the present invention, Verticillium lecanii and aconite alkaloid compounded pesticides has three kinds of formulations, i.e. encapsulated form, liquid preparation and compound powder.
At compounded pesticides is encapsulated form, and bacillus thuringiensis fermentation product and Verticillium lecanii fermentation product are zymotic fluid in its main ingredient, and each component content is by weight percentage,
Main ingredient 49.5-49.8%,
Auxiliary agent: emulsifier 0.2-0.5%, active substance protectant 0.1-0.3%, capsule material 49.5-49.8% and antioxidant 0.1-0.2%,
The capsule material mixture that the capsule material is selected from single capsule material or is made up of main capsule material and secondary capsule material, single capsule material is methylcellulose or polyamide, and main capsule material and secondary capsule material are 1: 1 sodium alginate and polylysine for the quality proportioning, the quality proportioning is 1: 1 sodium alginate and a shitosan, the quality proportioning is 1: 1 sodium alginate and a chitosan, the quality proportioning is 1: 1 gum Arabic and a gelatin, the quality proportioning is 1: 1 pectin and a gelatin, the quality proportioning is 1: 1 corn starch and a polyethylene glycol, the quality proportioning is 1: 1 sodium carboxymethylcellulose and a gelatin, the quality proportioning is that 1: 1 sodium alginate and albumin or quality proportioning are 1: 1 sodium alginate and barium chlorides;
Antioxidant is selected from a kind of in vitamin C, sodium sulphite or the sodium hydrogensulfite;
The encapsulated form preparation method is:
A, get single capsule material solution 121 ℃ of autoclavings 25 minutes, auxiliary agent ultraviolet sterilization 40 minutes, again with the bacillus thuringiensis zymotic fluid of 49.5-49.8%, Verticillium lecanii zymotic fluid and aconite alkaloid mixture are dispersed in the single capsule material solution of 49.5-49.8%, the emulsifier that adds 0.2-0.5% then sequentially, the active substance protectant of 0.1-0.3% and the antioxidant of 0.1-0.2%, fully stirring and evenly mixing got mixed liquor in 2 hours, the extruder that transfers to 0.01 micron with the aperture is evenly clamp-oned above-mentioned mixed liquor in the curing agent of 10 times of volumes of pre-preparation product population (w/v), solidify after 20-30 minute, get the gel grain after the isolated by filtration;
B, it was suspended in the capsule core material liquefier (w/v) of 10 times of volumes of pre-preparation product population liquefaction 24 hours, the pH value of capsule core material liquefier is 6.8-7.4, isolated by filtration, with twice of aseptic water washing, with after accounting for pre-preparation product population percentage by weight and being 0.2% lubricant disperse particles, normal temperature is dry down, gets product;
As choose capsule material mixture, in above-mentioned A step, then need bacillus thuringiensis zymotic fluid, Verticillium lecanii zymotic fluid and the aconite alkaloid mixture of 49.5-49.8% are dispersed in the main capsule material solution of 49.5-49.8%, after making the gel grain, the gel grain was suspended 60 minutes in secondary capsule material solution, carry out the B step then;
To be selected from concentration be 65% Na to curing agent among the above-mentioned preparation method
2SO
4Solution, concentration are 1.5% CaCl
2Solution, concentration are that 36% formalin, concentration are that 5% acetum or concentration are a kind of in 50% the sodium chloride solution; The capsule core material liquefier is selected from the pH value and is the neutral salt solution of 6.8-7.4, i.e. SAS, sodium chloride solution or calcium chloride solution, and lubricant is selected from talcum powder or dolomol.
Explain: curing agent, capsule core material liquefier and lubricant all belong to the material of auxiliary micro-capsule preparation, so do not account for the percentage composition of desiring prepared product.
Encapsulated form product advantage: safety non-toxic, long shelf-life, degeneration-resistant border ability is strong, control efficiency good.
At compounded pesticides is liquid preparation, and bacillus thuringiensis fermentation product and Verticillium lecanii fermentation product are zymotic fluid in the main ingredient, and each component content is by weight percentage,
Main ingredient 88.78-93.49%,
Auxiliary agent: active substance protectant 0.1-0.5%, emulsifier 0.3-0.5%, antioxidant 0.1-0.2%, uv-protector 0.01-0.02%, suspending agent 5-8%, antifreezing agent 1-2%;
Antioxidant is selected from a kind of in vitamin C, sodium sulphite or the sodium hydrogensulfite;
Uv-protector is selected from a kind of in fluorescein sodium, Tea Polyphenols, aesculin, folic acid, acridine yellow, the Congo red or methyl green;
Suspending agent is selected from a kind of in gumwater, glycerine, starch slurry, sodium alginate soln, methocel solution or the diatomite;
Antifreezing agent is selected from ethylene glycol or glycerine;
The liquid preparation method for making is: with suspending agent; uv-protector and antioxidant were 121 ℃ of autoclavings 25 minutes; other auxiliary agent ultraviolet irradiation sterilization 40 minutes; again with 88.78-93.49% bacillus thuringiensis zymotic fluid; 1: 1: 0.1 mixture of Verticillium lecanii zymotic fluid and aconite alkaloid and 0.01-0.02% uv-protector; 0.1-0.5% active substance protectant; the 0.3-0.5% emulsifier; 5-8% suspending agent and 0.1-0.2% antioxidant add in the mixing machine sequentially; with 150 rev/mins speed stirring and evenly mixings 60 minutes; add the 1-2% antifreezing agent again; continue stirring and evenly mixing 30 minutes, and promptly got the liquid preparation of transparent and homogeneous.
Liquid preparation product advantage: production technology is simple, product and water can with arbitrarily than mix, easy to use, pollution-free, control efficiency is good.
At compounded pesticides is compound powder, and the bacillus thuringiensis fermentation product is that solid is its zymotic fluid centrifugal sediment in the main ingredient, and the Verticillium lecanii fermentation product is the conidial powder of solid culture; Each component content is by weight percentage,
Main ingredient 75-85%,
Auxiliary agent: emulsifier 0.3-0.5%, active substance protectant 0.1-0.2%, uv-protector 0.01-0.02%, filler 14.59-24.28%;
The preparation method of Verticillium lecanii conidial powder is: the Verticillium lecanii spore that covers with in the potato medium with a small amount of aseptic kaolin wash-out, and is stripped down together with lawn, and air drying under aseptic condition is pulverized, and crosses 100 mesh sieves promptly;
Uv-protector is selected from a kind of in fluorescein sodium, Tea Polyphenols, aesculin, folic acid, acridine yellow, the Congo red or methyl green;
Filler is selected from a kind of in diatomite, microcrystalline cellulose, pregelatinized starch, sodium alginate, the calcium sulphate, perhaps be selected from the mixture of clay and sucrose, perhaps be selected from the mixture of maltose and dextrin, the quality proportioning of clay and sucrose is 10: 1, and the quality proportioning of maltose and dextrin is 1: 8.
Pregelatinized starch is purchased in Qufu City sky profit pharmaceutic adjuvant Co., Ltd.
The compound powder method for making is: auxiliary agent is crossed 100 mesh sieves in advance; ultraviolet irradiation sterilization 40 minutes; 1: 1: 0.1 mixture of centrifugal sediment, Verticillium lecanii conidial powder and the aconite alkaloid of 75-85% bacillus thuringiensis zymotic fluid is placed mixing machine; speed stirring and evenly mixing with 60 rev/mins; successively add 0.01-0.02% uv-protector, 0.1-0.2% active substance protectant, 0.3-0.5% emulsifier and 14.59-24.28% filler while stirring; again with 150 rev/mins the abundant mixing of speed 90 minutes; cross 100 mesh sieves, get product.
Compound powder product advantage: production technology is simple, cost is low, long shelf-life, storing convenient, control efficiency is good.
More than in three kinds of formulations emulsifier be selected from a kind of in sodium hexadecyl sulfate, lauryl sodium sulfate, Polyoxyethylene Sorbitan Monooleate, polyglycerol ester, polyglycereol linoleic acid one ester or the triethanolamine; The active substance protectant is selected from a kind of in sodium alginate, skim milk, L-serine, sodium dihydrogen phosphate, molasses, sucrose or the Tween-80.
The centrifugal sediment of bacillus thuringiensis zymotic fluid is a routine fashion: with bacillus thuringiensis zymotic fluid with 4000 rev/mins speed centrifugal 20 minutes, the supernatant that inclines, promptly.
Bacillus thuringiensis (bacillus thuringiensis) is called for short Bt, it is output maximum on the our times, a quasi-microorganism insecticide that is most widely used, account for the 70% above share in whole biopesticide market, China starts from 1958 to the research of bacillus thuringiensis, now existing tens manufacturers, annual production reaches more than 30,000 tons, product forms also develops into liquor by single pulvis, novel form such as emulsion and wetting powder, bacillus thuringiensis plays an important role in the green control of damage by disease and insect, but its desinsection speed is slow, be subject to such environmental effects, lasting period is short, insecticidal spectrum is narrow, because be by the stomach poison function desinsection, so to aphid, the perverse suction formula of trialeurodes vaporariorum insect does not almost have effect, and domestic present production cost is still too high, make up novel bacterial also just like stability by gene engineering, the all multifactor restrictions of safety make its further application development be subjected to very big influence.
Verticillium lecanii (vertillum lecanni zimmermanv viegas) is the class fungus insecticide that last century, the seventies was found, be all insect pathogenic fungus very widely of a kind of parasitic scope and geographical distribution, its main parasitic is in Homoptera insect, aphid particularly, aleyrodid and a red-spotted lizard class, be mainly used in anti-eliminating aphis and the greenhouse whitefly class pest abroad at present, it is wide that it has prevention and treatment range, lasting period is long, free from environmental pollution, to people and animals and natural enemies security, can use synergy with most chemical insecticides, the advantage of spread spectrum effect, and can cause that under the adapt circumstance condition insect cause of disease disease is popular, reach the purpose of Sustainable Control pest population, also can be used as the bactericide of plant disease, be a kind of biological and ecological methods to prevent plant disease, pests, and erosion exploitation bacterium that has a development potentiality, abroad existing a lot of national registrations multiple commodity.Its weak point is: desinsection speed is slower when using separately, and generally 5-6 talent sees a large amount of polypide death after dispenser, because of it is the parasitic formula desinsection of contact, so the polypide that medicine is failed to be sprayed onto does not have killing action; Shelf life of products is short, is active bacterium spore because mainly contain effective constituent, preserves difficulty relatively; Environmental factor is bigger to its influence, and especially temperature, humidity are the principal elements that influences its pathogenicity, and only spore could be sprouted under the condition of thermophilic, high humidity (25-30 ℃, more than the RH80%), and the host is caused a disease.Domestic still do not have its registration goods at present and occur, and do not have the large scale application report yet.
Aconite alkaloid (Aconitine) is the effective constituent that mainly contains of iron staff hammer (Aconitum pendulum Busch) root, stem, and the each province, northwest extensively distributes, and each province and city all have the medicinal material standard and put down in writing, and it is hot in nature, bitter is hot, very toxic.Have activating blood and removing stasis, dispel rheumatism, the effect of alleviating pain and detumescence, but its piece root has severe toxicity.At present, domestic main as people's medicine, but it also is the high-quality insecticide, especially phosphorus wing order, coleopteran pest there is special efficacy, its advantage: (aconite alkaloid also is the main active ingredient of the first-class various plants of monkshood to wide material sources, also available cell suspension cultures method obtains, thereby has overcome the sustainability problem of resource), desinsection speed is fast, virulence strong, long shelf-life, residual low.Shortcoming: the lasting period is short, during high concentration people and animals is had harm, sucks excessive meeting and causes poisoning, severe patient causes death, Zhang Hongli people utilizes Radix aconiti szechenyiani extract to make the rodenticide proof has accumulation intoxicating phenomenon to small white mouse, belongs to poisonous pesticide, thereby has limited its development at pesticide field.
Aconite alkaloid does not have influence (that is: do not reduce the sprouting of Verticillium lecanii spore and the ability of dip-dye insect, do not influence the normal growth of bacillus thuringiensis yet) to two kinds of built reactive bacterium, and is adjuvant used to the person poultry safety.
The present invention has overcome the shortcoming of its each single agent, and three kinds of effective single agent can produce higher synergistic effect, have widened insecticidal spectrum, improve desinsection speed and insecticidal effect, prolonged the lasting period, reduced toxicity, production, agricultural cost reduce, and production technology is simple, and raw material sources are extensive, safety and environmental protection, formulation is various, thereby degeneration-resistant border ability strengthens, and application is widened, and has very high exploitation value and vast potential for future development.
Compounded pesticides of the present invention can be prevented and treated the various plants insect, especially to Homoptera, Lepidoptera, coleoptera pilosity and the insect of refractory has lasting control effect, is the complex biological pesticidal preparations of a kind of efficient, environmental protection, safety.
The present invention can continue until the 15 day to the deadly peak period of Trialeurodes vaporariorum Westwood and cabbage caterpillar since second day, and preventive effect can reach more than 90%, and the shelf-life is more than 1.5 years.
Embodiment
The following examples can make those skilled in the art more fully understand the present invention, but do not limit the present invention in any way.
Main bacteria seed and source: bacillus thuringiensis is that CGMCC1.767 and CGMCC1.1012 were by 1: 1 mixture; Verticillium lecanii CGMCC3.4505 is all available from Chinese microorganism strain preservation center.
Aconite alkaloid is hammered ethanol extract into shape from iron staff in the main ingredient, the preparation method of iron staff hammer ethanol extract is, the iron staff hammer root of drying is ground into 60 order fine powders, with 95% ethanol serves as to extract solvent, the volume of ethanol is three times of iron staff hammer fine powder volume, place altogether and shake bottle jolting extraction 120 hours, filter, filter residue extracted 120 hours with the ethanol solvent jolting of the long-pending amount of triploid again, filter, filter residue merges all filtrates with solvent washing 3 times, decompression recycling ethanol promptly gets the iron staff hammer ethanol extract that contains total aconite alkaloid 82.3%.
Embodiment 1-11: the composite encapsulated form of bacillus thuringiensis, Verticillium lecanii and aconite alkaloid.
At compounded pesticides is encapsulated form, and bacillus thuringiensis fermentation product and Verticillium lecanii fermentation product are zymotic fluid in its main ingredient, and each component content sees Table one and table two respectively.
The encapsulated form preparation method is:
A, get single capsule material solution 121 ℃ of autoclavings 25 minutes, auxiliary agent ultraviolet sterilization 40 minutes, again with the bacillus thuringiensis zymotic fluid, Verticillium lecanii zymotic fluid and aconite alkaloid mixture are dispersed in the single capsule material solution, add emulsifier then sequentially, active substance protectant and antioxidant, fully stirring and evenly mixing got mixed liquor in 2 hours, the extruder that transfers to 0.01 micron with the aperture is evenly clamp-oned above-mentioned mixed liquor in the curing agent of 10 times of volumes of pre-preparation product population (w/v), solidify after 20-30 minute, get the gel grain after the isolated by filtration;
B, it was suspended in the capsule core material liquefier (w/v) of 10 times of volumes of pre-preparation product population liquefaction 24 hours, the pH value of capsule core material liquefier is 6.8-7.4, isolated by filtration, with twice of aseptic water washing, with after accounting for pre-preparation product population percentage by weight and being 0.2% lubricant disperse particles, normal temperature is dry down, gets product;
As choose capsule material mixture, in above-mentioned A step, then need bacillus thuringiensis zymotic fluid, Verticillium lecanii zymotic fluid and aconite alkaloid mixture are dispersed in the main capsule material solution, after making the gel grain, the gel grain was suspended 60 minutes in secondary capsule material solution, carry out the B step then;
To be selected from concentration be 65% Na to curing agent among the above-mentioned preparation method
2SO
4Solution, concentration are 1.5% CaCl
2Solution, concentration are that 36% formalin, concentration are that 5% acetum or concentration are a kind of in 50% the sodium chloride solution; The capsule core material liquefier is selected from the pH value and is the neutral salt solution of 6.8-7.4, i.e. SAS, sodium chloride solution or calcium chloride solution, and lubricant is selected from talcum powder or dolomol.
Explain: curing agent, capsule core material liquefier and lubricant all belong to the material of auxiliary micro-capsule preparation, so do not account for the percentage composition of desiring prepared product.If 50 kilograms of wet micro-capsules of desire preparation, by micro-capsule yield 80.6%, the total amount that then needs to feed intake is 62.03 kilograms.
Encapsulated form product advantage: safety non-toxic, long shelf-life, degeneration-resistant border ability is strong, control efficiency good.
The composite microcapsules Example formulations of table one, bacillus thuringiensis, Verticillium lecanii and aconite alkaloid amount
The composite microcapsules Example formulations of table two, bacillus thuringiensis, Verticillium lecanii and aconite alkaloid amount
Note: 80 kilograms of finished products of pre-preparation.
Embodiment 12-17, bacillus thuringiensis, Verticillium lecanii and aconite alkaloid compositional liquor body preparation
At compounded pesticides is liquid preparation, and bacillus thuringiensis fermentation product and Verticillium lecanii fermentation product are zymotic fluid in the main ingredient, and each component content sees Table three.
The liquid preparation method for making is: with suspending agent, uv-protector and antioxidant 121 ℃ of autoclavings 25 minutes; other auxiliary agent ultraviolet irradiation sterilization 40 minutes; add 1: 1: 0.1 mixture of bacillus thuringiensis zymotic fluid, Verticillium lecanii zymotic fluid and aconite alkaloid and uv-protector, active substance protectant, emulsifier, suspending agent and antioxidant in the mixing machine sequentially again; with 150 rev/mins speed stirring and evenly mixings 60 minutes; add the 1-2% antifreezing agent again; continue stirring and evenly mixing 30 minutes, and promptly got the liquid preparation of transparent and homogeneous.
Liquid preparation product advantage: production technology is simple, product and water can with arbitrarily than mix, easy to use, pollution-free, control efficiency is good.
Table three, bacillus thuringiensis, Verticillium lecanii and aconite alkaloid compositional liquor body preparation Example formulations amount
Note: 80 kilograms of finished products of pre-preparation.
Embodiment 18-24, bacillus thuringiensis, Verticillium lecanii and aconite alkaloid compound powder
At compounded pesticides is compound powder, and the bacillus thuringiensis fermentation product is that solid is its zymotic fluid centrifugal sediment in the main ingredient, and the Verticillium lecanii fermentation product is the conidial powder of solid culture; Each component content sees Table four.
The preparation method of Verticillium lecanii conidial powder is: in the 500ml blake bottle with the Verticillium lecanii spore 200ml that covers with in the potato medium with the aseptic kaolin wash-out of 1-2g, and strip down together with lawn, air drying under aseptic condition is pulverized, and crosses 100 mesh sieves promptly;
Pregelatinized starch is purchased in Qufu City sky profit pharmaceutic adjuvant Co., Ltd.
The compound powder method for making is: auxiliary agent is crossed 100 mesh sieves in advance; ultraviolet irradiation sterilization 40 minutes; 1: 1: 0.1 mixture of centrifugal sediment, Verticillium lecanii conidial powder and the aconite alkaloid of bacillus thuringiensis zymotic fluid is placed mixing machine; speed stirring and evenly mixing with 60 rev/mins; successively add uv-protector, active substance protectant, emulsifier and filler while stirring; with 150 rev/mins the abundant mixing of speed 90 minutes, cross 100 mesh sieves again, get product.
The centrifugal sediment of bacillus thuringiensis zymotic fluid is a routine fashion: with bacillus thuringiensis zymotic fluid with 4000 rev/mins speed centrifugal 20 minutes, the supernatant that inclines, promptly.Compound powder product advantage: production technology is simple, cost is low, long shelf-life, storing convenient, control efficiency is good.
Table four, bacillus thuringiensis, Verticillium lecanii and aconite alkaloid compound powder Example formulations amount
Note: 80 kilograms of finished products of pre-preparation.
Among the above embodiment, it is long that embodiment 5,11,16 and 23 makes shelf life of products, and control efficiency is good, compares better with other embodiment with formulation.
Field efficiency test to Trialeurodes vaporariorum Westwood
In the greenhouse of cultivation poinsettia; poinsettia plant 560 strains of choosing serious biennial of Trialeurodes vaporariorum Westwood harm and having bloomed; be divided into 7 rectangle sub-districts; five is the test drug sub-district; a positive control plot; one is the blank sub-district; add guard row between per two sub-districts; every small plot experiment plant 80 strains, every sub-district is got 20 strains at random, whenever selects good strains in the field for seed three (from plant; in; following three positions are chosen) the more blade of borer population writes down the basic borer population and the mark of listing; then; spray experimental drug simultaneously; contrast medicine and clear water spray from the blade two sides, with the blade degree of being that drips.A few days ago sheltering from heat or light, preserving moisture after the dispenser, and after dispenser live borer population and have or not poisoning of the 3rd, five, seven, 15 day observed and recorded.Test of pesticide effectiveness method in the sample shelf-life is the same, and result of the test sees Table five.
Table five, bacillus thuringiensis, Verticillium lecanii and aconite alkaloid compounded pesticides are to the field efficacy of Trialeurodes vaporariorum Westwood
Note: the correction preventive effect in the table in the shelf-life is the result of investigation in five days after the dispenser at that time.The compositional liquor body preparation is pressed embodiment 16 preparations, and compound powder is pressed embodiment 23 preparations, and all the other mono-component liquid pharmaceutical formulations are identical with embodiment 16, only changed main ingredient.
Bacillus thuringiensis, Verticillium lecanii and aconite alkaloid compounded pesticides are to the field efficiency test of cabbage caterpillar
In the greenhouse of cultivation Chinese cabbage, choose serious plant 420 strains of cabbage caterpillar harm, be divided into 7 rectangle sub-districts; five is the test drug sub-district, a positive control plot, and one is the blank sub-district; establish guard row between per two sub-districts; every small plot experiment plant 60 strains, every sub-district is got 30 strains at random, writes down the every strain basis borer population and the mark of listing; then; spray experimental drug, contrast medicine and clear water simultaneously, from plant up and down complete stool spray, with the blade degree of being that drips.A few days ago sheltering from heat or light, preserving moisture after the dispenser, and after dispenser live borer population and have or not poisoning of the 3rd, five, seven, 15 day observed and recorded.Test of pesticide effectiveness method in shelf-life is the same, and result of the test sees Table six.
Table six, bacillus thuringiensis, Verticillium lecanii and aconite alkaloid compounded pesticides are to the field efficacy of cabbage caterpillar
Note: the correction preventive effect in the table in the shelf-life is the result of investigation in five days after the dispenser at that time.The compositional liquor body preparation is pressed embodiment 5 preparations, and compound powder is pressed embodiment 23 preparations, and all the other mono-component liquid pharmaceutical formulations are identical with embodiment 16, only changed main ingredient.
According to embodiment as can be known, complex preparation of the present invention is compared with positive control with single agent of its composition and is had the following advantages:
(1) enlarges insecticidal spectrum, reduced toxicity.Bacillus thuringiensis is mainly to kill Lepidoptera, coleopteran pest, and is invalid substantially to homoptera pest.Verticillium lecanii belongs to the broad spectrum disinfestation fungi, but it has special efficacy to homoptera pest, especially aphid and Trialeurodes vaporariorum Westwood is had special control efficiency.Aconitine is all effective to Lepidoptera and homoptera pest, but people and animals is had harm during its high concentration.After the three formed complex preparation, can effectively prevent and treat Homoptera, Lepidoptera and coleopteran pest simultaneously, also avoid its toxic hazard.
The biological medicine of the present invention is effective, and avirulence can substitute virose chemical medicine Ah. pyridine worm fully and narrow or sumicidin.
(2) improved desinsection speed.Bacillus thuringiensis, Verticillium lecanii began at the 5th day the deadly peak period of pest control, and the deadly peak period of its complex preparation can continue until the 7th day since second day.(3) improved insecticidal toxicity.Three components have higher synergistic effect in the complex preparation, compare with each one pack system, and virulence has improved 20%--50%.(4) prolonged the longevity of residure.Single agent in the time of the 15 day relative control effect just reduce to below 60%, but complex preparation after 15 days relative control effect still more than 90%.(5) prolonged the shelf-life.By composite microcapsules, liquid preparation and the compound powder of formulation of the present invention, its shelf-life, all more than 1.5 years, activity also improved.(6) cost reduces.Compare with single component formulations, delayed the generation of pest resistance to insecticide, production cost has reduced more than 30%, and agricultural cost has reduced 30%-50%.(7) environmental protection, to the person poultry safety.Complex preparation is noresidue in use, with the maximum method of once limiting the quantity of, with 2 * 10
11The amount of individual gemma (spore)/kg body weight to the acute oral of small white mouse and percutaneous toxicity experiment showed, do not have cause death, sensitization and bad reaction.
Claims (9)
1. the preparation method of a bacillus thuringiensis, Verticillium lecanii and aconite alkaloid compounded pesticides is characterized in that:
With main ingredient and additive mixture, the percentage by weight of main ingredient is that the percentage by weight of 50%-94%, auxiliary agent is 6-50%; Main ingredient is that weight ratio is the mixture of 1: 1: 0.1 bacillus thuringiensis (Bacillus thuringiensis) fermentation product, Verticillium lecanii (Verticillium lecanii) fermentation product and aconite alkaloid, and auxiliary agent comprises emulsifier and active substance protectant.
2. bacillus thuringiensis according to claim 1, the preparation method of Verticillium lecanii and aconite alkaloid compounded pesticides, it is characterized in that: aconite alkaloid is hammered ethanol extract into shape from iron staff in the main ingredient, the preparation method of iron staff hammer ethanol extract is, the iron staff hammer root of drying is ground into 60 order fine powders, with 95% ethanol serves as to extract solvent, the volume of ethanol is three times of iron staff hammer fine powder volume, place altogether and shake bottle jolting extraction 120 hours, filter, filter residue extracted 120 hours with the ethanol solvent jolting of the long-pending amount of triploid again, filter, filter residue washs 3 times with solvent, merge all filtrates, decompression recycling ethanol promptly gets the iron staff hammer ethanol extract that contains total aconite alkaloid 82.3%.
3. the preparation method of bacillus thuringiensis according to claim 1 and 2, Verticillium lecanii and aconite alkaloid compounded pesticides is characterized in that: emulsifier is selected from a kind of in sodium hexadecyl sulfate, lauryl sodium sulfate, Polyoxyethylene Sorbitan Monooleate, polyglycerol ester, polyglycereol linoleic acid one ester or the triethanolamine; The active substance protectant is selected from a kind of in sodium alginate, skim milk, L-serine, sodium dihydrogen phosphate, molasses, sucrose or the Tween-80.
4. the preparation method of bacillus thuringiensis according to claim 3, Verticillium lecanii and aconite alkaloid compounded pesticides, it is characterized in that: compounded pesticides is an encapsulated form, bacillus thuringiensis fermentation product and Verticillium lecanii fermentation product are zymotic fluid in the main ingredient, auxiliary agent also comprises capsule material and antioxidant, each component content is by weight percentage
Main ingredient 49.5-49.8%, emulsifier 0.2-0.5%, active substance protectant 0.1-0.3%, capsule material 49.5-49.8% and antioxidant 0.1-0.2%,
The capsule material mixture that the capsule material is selected from single capsule material or is made up of main capsule material and secondary capsule material, single capsule material is methylcellulose or polyamide, and main capsule material and secondary capsule material are 1: 1 sodium alginate and polylysine for the quality proportioning, the quality proportioning is 1: 1 sodium alginate and a shitosan, the quality proportioning is 1: 1 sodium alginate and a chitosan, the quality proportioning is 1: 1 gum Arabic and a gelatin, the quality proportioning is 1: 1 pectin and a gelatin, the quality proportioning is 1: 1 corn starch and a polyethylene glycol, the quality proportioning is 1: 1 sodium carboxymethylcellulose and a gelatin, the quality proportioning is that 1: 1 sodium alginate and albumin or quality proportioning are 1: 1 sodium alginate and barium chlorides; Antioxidant is selected from a kind of in vitamin C, sodium sulphite or the sodium hydrogensulfite;
The preparation method is:
A, get single capsule material solution 121 ℃ of autoclavings 25 minutes, auxiliary agent ultraviolet sterilization 40 minutes, again with the bacillus thuringiensis zymotic fluid of 49.5-49.8%, Verticillium lecanii zymotic fluid and aconite alkaloid mixture are dispersed in the single capsule material solution of 49.5-49.8%, the emulsifier that adds 0.2-0.5% then sequentially, the active substance protectant of 0.1-0.3% and the antioxidant of 0.1-0.2%, fully stirring and evenly mixing got mixed liquor in 2 hours, the extruder that transfers to 0.01 micron with the aperture is evenly clamp-oned above-mentioned mixed liquor in the curing agent of 10 times of volumes of pre-preparation product population, solidify after 20-30 minute, get the gel grain after the isolated by filtration;
B, it was suspended in the capsule core material liquefier of 10 times of volumes of pre-preparation product population liquefaction 24 hours, the pH value of capsule core material liquefier is 6.8-7.4, isolated by filtration, with twice of aseptic water washing, with after accounting for pre-preparation product population weight percentage and being 0.2% lubricant disperse particles, normal temperature is dry down, gets product;
As choose capsule material mixture, in above-mentioned A step, then need bacillus thuringiensis zymotic fluid, Verticillium lecanii zymotic fluid and the aconite alkaloid mixture of 49.5-49.8% are dispersed in the main capsule material solution of 49.5-49.8%, after making the gel grain, the gel grain was suspended 60 minutes in secondary capsule material solution, carry out the B step then;
To be selected from concentration be 65% Na to curing agent among the above-mentioned preparation method
2SO
4Solution, concentration are 1.5% CaCl
2Solution, concentration are that 36% formalin, concentration are that 5% acetum or concentration are a kind of in 50% the sodium chloride solution; The capsule core material liquefier is selected from the pH value and is the neutral salt solution of 6.8-7.4, i.e. SAS, sodium chloride solution or calcium chloride solution, and lubricant is selected from talcum powder or dolomol.
5. the bacillus thuringiensis that makes according to the described preparation method of claim 4, Verticillium lecanii and aconite alkaloid compounded pesticides encapsulated form.
6. the preparation method of bacillus thuringiensis according to claim 3, Verticillium lecanii and aconite alkaloid compounded pesticides; it is characterized in that: compounded pesticides is a liquid preparation; bacillus thuringiensis fermentation product and Verticillium lecanii fermentation product are zymotic fluid in the main ingredient; auxiliary agent also comprises antioxidant, uv-protector, suspending agent and antifreezing agent; each component content is by weight percentage
Main ingredient 88.78-93.49%, active substance protectant 0.1-0.5%, emulsifier 0.3-0.5%, antioxidant 0.1-0.2%, uv-protector 0.01-0.02%, suspending agent 5-8%, antifreezing agent 1-2%;
Antioxidant is selected from a kind of in vitamin C, sodium sulphite or the sodium hydrogensulfite; Uv-protector is selected from a kind of in fluorescein sodium, Tea Polyphenols, aesculin, folic acid, acridine yellow, the Congo red or methyl green; Suspending agent is selected from a kind of in gumwater, glycerine, starch slurry, sodium alginate soln, methocel solution or the diatomite; Antifreezing agent is selected from ethylene glycol or glycerine;
With suspending agent; uv-protector and antioxidant were 121 ℃ of autoclavings 25 minutes; other auxiliary agent ultraviolet irradiation sterilization 40 minutes; again with 88.78-93.49% bacillus thuringiensis zymotic fluid; 1: 1: 0.1 mixture of Verticillium lecanii zymotic fluid and aconite alkaloid and 0.01-0.02% uv-protector; 0.1-0.5% active substance protectant; the 0.3-0.5% emulsifier; 5-8% suspending agent and 0.1-0.2% antioxidant add in the mixing machine sequentially; with 150 rev/mins speed stirring and evenly mixings 60 minutes; add the 1-2% antifreezing agent again; continue stirring and evenly mixing 30 minutes, and promptly got the liquid preparation of transparent and homogeneous.
7. the bacillus thuringiensis that makes according to the described preparation method of claim 6, Verticillium lecanii and aconite alkaloid compounded pesticides liquid preparation.
8. the preparation method of bacillus thuringiensis according to claim 3, Verticillium lecanii and aconite alkaloid compounded pesticides, it is characterized in that: compounded pesticides is a compound powder, the bacillus thuringiensis fermentation product is its zymotic fluid centrifugal sediment in the main ingredient, and the Verticillium lecanii fermentation product is the conidial powder of solid culture; Auxiliary agent also comprises uv-protector and filler, and each component content is by weight percentage,
Main ingredient 75-85%, emulsifier 0.3-0.5%, active substance protectant 0.1-0.2%, uv-protector 0.01-0.02%, filler 14.59-24.28%;
Uv-protector is selected from a kind of in fluorescein sodium, Tea Polyphenols, aesculin, folic acid, acridine yellow, the Congo red or methyl green; Filler is selected from a kind of in diatomite, microcrystalline cellulose, pregelatinized starch, sodium alginate, the calcium sulphate, perhaps be selected from the mixture of clay and sucrose, perhaps be selected from the mixture of maltose and dextrin, the quality proportioning of clay and sucrose is 10: 1, and the quality proportioning of maltose and dextrin is 1: 8;
The preparation method of Verticillium lecanii conidial powder is: the Verticillium lecanii spore that covers with in the potato medium with a small amount of aseptic kaolin wash-out, and is stripped down together with lawn, and air drying under aseptic condition is pulverized, and crosses 100 mesh sieves promptly;
Auxiliary agent is crossed 100 mesh sieves in advance; ultraviolet irradiation sterilization 40 minutes; 1: 1: 0.1 mixture of centrifugal sediment, Verticillium lecanii conidial powder and the aconite alkaloid of 75-85% bacillus thuringiensis zymotic fluid is placed mixing machine; speed stirring and evenly mixing with 60 rev/mins; successively add 0.01-0.02% uv-protector, 0.1-0.2% active substance protectant, 0.3-0.5% emulsifier and 14.59-24.28% filler while stirring; again with 150 rev/mins the abundant mixing of speed 90 minutes; cross 100 mesh sieves, get product.
9. the bacillus thuringiensis that described according to Claim 8 preparation method makes, Verticillium lecanii and aconite alkaloid compounded pesticides compound powder.
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| CN110959635A (en) * | 2019-11-27 | 2020-04-07 | 李时平 | Insect-killing and insect-preventing agent |
| CN115251085A (en) * | 2022-07-14 | 2022-11-01 | 山东省农业科学院 | Insecticidal fungus live spore water suspending agent and preparation method thereof |
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