CN105076216A - Microbial combined insecticide and preparation method thereof - Google Patents
Microbial combined insecticide and preparation method thereof Download PDFInfo
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- CN105076216A CN105076216A CN201410210981.2A CN201410210981A CN105076216A CN 105076216 A CN105076216 A CN 105076216A CN 201410210981 A CN201410210981 A CN 201410210981A CN 105076216 A CN105076216 A CN 105076216A
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Abstract
The invention discloses a microbial combined insecticide and its preparation method. The microbial combined insecticide is a concentrated cenobium preparation prepared from two or three strains selected from 40-60 parts by weight of fungi, 15-25 parts by weight of actinomycetes and/or 20-40 parts by weight of bacteria. The preparation method comprises the following steps: respectively culturing various strains in a routine way to obtain strain culture pure liquids; enlarged culture of bacteria solutions: mixing the strain culture pure liquids in proportion to prepare mixed bacteria solutions; mixing the mixed bacteria solutions according to the weight ratio; culturing with a culture solution and fermenting to obtain a product with high concentration of 2 billions/ml and a solid product. The microbial combined insecticide provided by the invention has the characteristic of poisoning different insects and insect species, has a strong poisoning effect on hundreds of pests such as scarab, mosquito, nematode, Lepidoptera, coleoptera, Hymenoptera and the like, has advantages of fast speed, and wide range and thorough poisoning. In comparison with insecticidal efficacy of an insecticide produced by the use of a single strain, insecticidal efficacy of the microbial combined insecticide is raised by 51% and above.
Description
Technical field
The present invention relates to a kind of microorganism insecticide, particularly relate to a kind of antimicrobial composition insecticide and preparation method thereof.
Background technology
Since twentieth century, world population increases very fast, requires also increasing to the total demand of food." the petrochemistry agricultural " of people's invention, chemical fertilizer and agricultural chemicals help the mankind to obtain abundance of food resource, facilitate the development and progress of the mankind.
At present, there are 7,500,000,000 populations in the whole world, will reach 9,000,000,000 after the year two thousand fifty, and the end of this century is close to the peak of 10,500,000,000 populations.So the mankind can increase considerably the requirement of food.The mankind to the dependence of " oil-agriculture " also in continuous reinforcement.
According to world's authoritative department statistics, the chemical substance of current whole world Prof. Du Yucang has more than 1,100 ten thousand kinds, also constantly increases with the quantity increasing by 1,000,000 kinds and 3 ~ 400,000,000 tons every year in recent years.Cause and there is material poisonous in a large number and harmful substance endangers global environment, ocean and air, the wide spectrum feature of chemical toxic substances and the technical development of chemicals Composite, too increase the difficulty of removing, degraded, decomposition, become the procephalon of the long-term hostile environment pollution disaster of the mankind.Family more than 1700 of existing insecticide factory of China, annual production about 350,000 tons of former medicines, process preparation 80 ~ 1,000,000 tons, pesticide active ingredient kind more than 40, product about more than 7000, but also the external highly toxic pesticide do not re-used that import is a large amount of, apply domestic soil.A large amount of pesticide producing out, while for human use, is poisoning the mankind oneself again.Particularly most of agricultural chemicals is the toxin agricultural chemicals with interior absorption, and toxicity is absorbed directly in the histocyte of crops stem, leaf, root, and enters fruit, Interior Seed, causes the very alert people of the permanence of murder by poisoning, intractable.
The use of agricultural chemicals and the relation of environmental protection; many scientists are arguing the infringement problem about reality and potentiality intensely; but there is a fact to be apparent; even press recommendation amount to use; some agricultural chemicals; particularly insecticide, really can retain considerable time and constitute the pollution to soil and water after a procedure.In some instances, although working concentration is little, it is also a serious threat.Such as, the aldrin of 1mg/kg is had also can to cause the death of 100% in the food of the every ration of quail.
Since half a century, the use of people to chemical synthetic pesticide is accustomed to, and bringing a well-known difficult problem thus: food, soil, underground water, rivers and lakes, ocean, air all receive the severe contamination of chemical residual, side effect is created to non-target pests and other biological simultaneously, cause the destruction of the ecosystem.The insect species of resistance and recovery ability is produced to chemical pesticide and quantity constantly increases.In addition, using and dealing with improperly due to synthesis highly toxic pesticide, many non-lethal events quite occur.
In view of many chemical synthetic pesticides can produce many side effects, many both developed and developing countries all constantly recognize the harm of the environmental pollution that chemical synthetic pesticide causes and some toxicological effect.Thus, microorganism and natural plants, in Development of Novel agricultural chemicals, become the key player that people pay close attention to gradually.
Due to above various reasons, chemical toxic substances, residue of pesticide toxin causes extreme influence to environment for human survival, utilize microorganism to kill off the insect pests so the present invention relates to, replace chemical pesticide, nontoxic to people and animals, not welding is a kind of safe biological method.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of antimicrobial composition insecticide and preparation method thereof, overcome the problem of prior art, have and can kill different insects and the characteristic of caste, to chafer, mosquito, nematode, Lepidoptera, the hundreds of such as coleoptera and Hymenoptera kind insect has that toxic action is strong, speed is fast, scope is wide and kill advantage thoroughly.
Technical problem to be solved by this invention is achieved through the following technical solutions:
A kind of antimicrobial composition insecticide is by the concentrated cenobium preparation being selected from fungi, 2 classes of actinomycetes and/or bacterium or 3 class bacterial classifications and being prepared into; Described parts by weight of component ratio is: fungi 40-60, actinomycetes 15-25 and/or bacterium 20-40; Described parts by weight of component ratio is preferably: fungi 50, actinomycetes 20 and/or bacterium 30.
Described mushroom, is selected from following 2 kinds or two or more bacterial classification respectively:
Fungi: stiff ACCC-30101 ← clay fertilizer institute of the Chinese Academy of Agricultural Sciences ← sand control institute of AS3.3675 ← Shan Xi Yu forest land district of Institute of Microorganism, Academia Sinica of (1) metarhiziumaunisopliae (metschn) sorok chafer edge, is separated from paranthrene tabaniformis larva; For anti-corn borer, chafer, clearwing moth etc.(2) Paecilomycesfarinosus (Dicks.) Brown turns ceramic route ACCC-30190 ← Chinese Academy of Agricultural Sciences's clay fertilizer institute ← Beijing Forestry University's force literary composition of presenting oneself before (a monarch) is given; Original number: 8601, control dendrolimus tabulaeformis.
(bright giving is wished by 3Paecilomyceslilacinus (Thom) Samson paecilomyces lilacinus ACCC-30677 ← tobacco research institute of Yunnan Province; Original number: ZML086; Locality: Yunnan; Separation source: root-knot nematode egg; Study as biological control.
Actinomycetes:
(1) Streptomycesanulatus (Beijingrinck1912) wdksman1953 garden ring streptomycete ACCC-41456 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00797; Metabolite has insecticidal activity; Separation source: soil; Be separated place: Lushi, henan five li of rivers.
(2) raw institute is planted in " Streptomycesculicidicus " Yanetal. streptomyces culicidicus ACCC-41132 ← institute of microbiology ← Shanghai, Guangzhou, Guangdong Province; Original number: GIM4.12MIK4.1; Kill mosquito larvae; Separation source: soil; Be separated place: Guangzhou.
(3) StreptomycesgriseolussubsphangzhouensisACCC-40075 ← Zhejiang Agriculture academy of sciences; Original number: S26; Product kills aphid element, very strong to aphid, two spotted spider mite and some lepidopterous larvae toxic action.
(4) StreptomycescalvusBackusetal., 1957 bald naked streptomycete ACCC-41451 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00743; Metabolite has insecticidal activity; Separation source: soil; Be separated place: Zhejiang dragon trip.
Bacterium:
(1) Bacillussubtilis (Enrenberg1835) cohn1872 bacillus subtilis ACCC-11088 ← Institute of Microorganism, Academia Sinica; AS1.0933; Control peanut nematodosis.
(2) BacillusSphaericusmeyerandNeide1904 Bacillus sphaericus ACCC-11096 ← Institute of Microorganism, Academia Sinica ← U.S.;=AS1.1672; Original number: 2115; Kill mosquito.
(3) BacilluslaterosporusLaubach bacillus laterosporus ACCC-11079 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute ← Microbe Inst., Chinese Academy of Sciences AS1.0864; Larvae can be killed; Produce lysozyme.
(4) Bacillusthuringienissubsp.AisawaiACCC-10016 ← Wuhan microbial pesticide factory; Original number: 096; Prepare antigen reference culture, poisoning lepidopterous insects.
(5) Bacillusthuringienissubsp.merrisoniACCC-10031 ← Wuhan microbial pesticide factory; Original number: 012; Poisoning lepidopterous insects.
(6) Bacillusthuringiensissubsp.galleriae bacillus thuringiensis,Bt galleria mellonella waxmoth subspecies ACCC-10027 ← Microbe Inst., Chinese Academy of Sciences; Poisoning cabbage caterpillar.
(7) Bacillusthuringiensissubsp.Israelensis Bacillus thuringiensis subsp. israelensis ACCC-10038 ← Chengdu Hua Hong industrial corporation ← Taian Shandong biological study institute ← World Health Organization (WHO); Kill mosquitoes larva, Jue
effect is better.
(8) Bacillusthuringienissubsp.thuringienisACCC-10035 ← China Agriculture Academe Fertilizer Institute; Original number: D25; Antiphagin poisoning corn borer; Separation source: spontaneons screening from HD-187 bacterial strain and obtaining.
(9) Bacillusthuringiensissuhsp.berliner bacillus thuringiensis,Bt Bai Shi mutation ACCC-10022 ← China Agriculture Academe Fertilizer Institute; Original number: D26; Control lepidopterous insects; Separation source: bacterium powder.
(10) Bacillusthuringiensissubsp.Dendrolimus bacillus thuringiensis,Bt pine another name for Sichuan Province subspecies ACCC-10062 ← Soviet Union; Poisoning pine caterpillars.
(11) Bacillusthuringienissubsp.sottoACCC-10033 ← Wuhan microbial pesticide factory ← Britain; Original number: 016; Prepare antigen reference culture, poisoning lepidopterous insects.
According to (0.5-1.5) between the pure liquid of each Spawn incubation of selected every class bacterial classification: (0.5-1.5) ratio mixes.
According to 1: 1 ratio mixing between the pure liquid of each Spawn incubation of selected every class bacterial classification.
Described preparation is every 2,000,000,000/milliliter high concentration product, and obtains fluid product and solid product further.
Present invention also offers described antimicrobial composition method for producing insecticide, comprise the following steps:
(1) by various bacterial classification respectively cellar culture be the pure liquid of Spawn incubation:
(2) expansion of bacterium liquid is cultivated: carry out being mixed and made into every class mixed bacteria liquid according to described ratio by between the pure liquid of each Spawn incubation of every class fungi, actinomycetes and/or bacterium; Again described every class Mixed Microbes is mixed in described parts by weight of component ratio; Culture fluid is cultivated, and fermentation obtains finished product preparation.
Described step (2) be by mixed bacteria liquid and culture fluid in 1: the ratio of (8-10) is cultivated, described culture fluid furnish component weight ratio is: analysis for soybean powder 1-1.5, dextrin 0.3-0.9, peptone 0.2-0.4, dipotassium hydrogen phosphate 0.1-0.3, calcium carbonate 0.3-0.7, magnesium sulfate 0.2-0.6, ammonium sulfate 0.03-0.07, soya-bean oil 0.1-0.3; PH value is 7.2.
Preferred described mixed bacteria liquid and culture fluid are cultivated in the ratio of 1: 9, and described culture fluid furnish component weight ratio is: analysis for soybean powder 1.2, dextrin 0.6, peptone 0.3, dipotassium hydrogen phosphate 0.2, calcium carbonate 0.5, magnesium sulfate 0.4, ammonium sulfate 0.05, soya-bean oil 0.2.
Described mixed bacteria liquid adds in the liquid medium after purification; Put into seeding tank, at temperature 30 DEG C ± 1 DEG C, stirring per hour 200 times, cultivates 36-48 hour; Microscopy every gram bacteria containing amount reaches 2,000,000,000/gram time, stop cultivating, check and be registered seed bacterium liquid without miscellaneous bacteria.By qualified bacterium liquid in the ratio of 5%, add liquid medium in fermentation tank, cultivate 30-45 hour, on inspection without miscellaneous bacteria, bacterium liquid obviously becomes gemma and crystal, and content reaches 2,000,000,000/gram, namely can be finished product fermentation product.
Described method also comprises the preparation of step (3) solid fungicide: the calcium carbonate adding 8-10% in finished product bacterium liquid, stirs 30 minutes; Plate-frame filtering; Stir 30 minutes, furnishing pulpous state; Spraying dry; Be pulverized and mixed evenly, obtain final product.
The present invention builds insecticide bacterial strain system, wherein Bacillus sphaericus BacillussphaericusTS-1 and Bacillus thuringiensis H4 is two kinds of important insect pathogenic bacterias, the former is suitable with external 1593-4 and T-M-1 bacterial strain to the toxicity of Culex pipiens pallens, and the latter has high virulence to armyworm, corn borer etc.Its preparation is existing merchandized handling abroad.Their toxicity all derives from toxalbumin specific to thalline, these toxalbumin are coded by chromosome or plasmid DNA, by Protoplast Fusion Technique, the different breeding bacterial strains of bacillus Pseudomonas are merged, the recombinant bacterial strain not only killing diptera but also kill lepidopterous larvae can be obtained.Obtain the stable fusion recombinant bacterial strain both killing lepidopterous larvae from bacillus thuringiensis,Bt, obtained successfully.
This insecticide is strong by killing ability, the concentrated cenobium of three classes, 18 microorganism fungus kind compositions that desinsection speed is fast, insecticidal range is wide, and this insecticide has liquid and solid two kinds, and the insecticide of insecticidal power ratio single culture improves effect and reaches 51%.
Described insecticidal range is wide, is because 18 bacterial classifications have main desinsection object, adopts the combination of integrated multifunction brute force can kill chafer, mosquito, nematode, Lepidoptera, the hundreds of such as Hymenoptera and coleoptera kind insect.Contrast with traditional single culture insecticide, killing ability is improved insecticidal power by single insecticide.
Have three mushrooms, 18 bacterial classifications in the insecticide bacterial classification system that the present invention builds, wherein: fungi has three, respectively effect is killed to insect chafer, pine tag electronics and larvae.Actinomycetes have four, have toxic action respectively to flood dragon worm and multiple lepidoptera pest.Bacterium has 11, and wherein three bacillus eight respectively have the bacillus thuringiensis,Bt of desinsection special efficacy, and they have the various pests such as poisoning corn borer, cabbage caterpillar, mosquito, nematode, Lepidoptera and diptera respectively.
The feature that bacterial classification of the present invention is selected is: one selects the strong bacterial classification of insect toxic action to obtain better poisoning ability and effect.Two select to have the bacterial classification killing different insect pest, to increase the broad spectrum disinfestation feature of insecticide.Three select different desinsection mushrooms, to obtain the interdependence of mushroom bacterial classification, increase the symbiosis effect of the spawn activity activity time stronger with maintenance.
The present invention relates to rising and insecticidal mechanism of antimicrobial composition insecticide:
The microorganisms such as fungi, bacterium, virus all can make insect obtain communicable disease.The situation of bacterium, fungi and virus infection insect is comparatively common, to the research of insect disease from beneficial insect silkworm and honeybee.Chinese medicine Cordyceps sinensis also by the insect bodies of fungi autoeciousness, but to the understanding of insect-borne infection cause of disease then development microbiological with modern age be associated.Until within 1835, begin know that the white muscardine of silkworm is caused by fungi white muscardine fungi.Shi Ao state scarab beetle (AnisoaustriacaHbst) Infective, for utilizing the early test of microbial control insect.Research that is bacillary to insect and viral infectious has been expanded to since 20 beginnings of the century.Research comparatively early has: locust coccobacillus is to the control of desert locust, and Bacillus popilliae is to the control of Japanese beetle first, and bacillus thuringiensis,Bt is to the control of corn borer, and nucleopolyhedrosis virus is to the controlling experiment etc. of military poison moth and diprion hercyniae.
After various microbial infection insect, insect often shows the abnormality in function and behavior, the symptom (symptom) of meaning, and the constructional change of polypide, the symptom (sign) of meaning.Both symptom and symptom are generically and collectively referred to as syndrome (syn-drome) (Steinhaus and Martigoni, 1970) altogether.Usually be there will be the multiple symptom such as action and abnormal reaction (show as slow in action, slow in reacting etc.), indigestion (show as heavyly to subtract, stops taking food, vomit, dysentery shape etc.), maldevelopment (poor growth, can not normally cast off a skin, pupate and emergence etc.), reproductive disorder (fecundity reduces, can not lay eggs), body colour change by the insect of infected by microbes to show, these symptom can be used as the foundation that discriminating insect under field conditions (factors) catches an illness.The syndrome that cause of disease of the same race causes generally has certain feature, but also can have difference in various degree under different environmental conditions.Generally speaking, because the polypide that fungal infection is lethal is withered or rigid; Die from the body tissue liquefaction of polyhedrosis virus, body body is easy to break; The polypide dying from bacteriosis is softened, and body fluid is thinning, the liquid flowed out from mouth and anus and the equal foul smelling of liquid reserved after breaking; Due to the polypide worm that different reason is lethal, belong to normal intestinal bacteria originally and also can breed rapidly, often beyond really lethal cause of disease, also disorganize can produce stench and cause the confusion in diagnosis.
The pathogenetic bacteria of insect generally can be divided into the two large classes forming gemma and do not form gemma, and according to the parasitism of insect being further divided into obligate and two facultative classes.Bucher (1960) by the characteristic of pathogenetic bacteria and required condition of causing a disease as infective dose, infect position, entomopathogenic bacterium, four classes be divided into the selectivity of host and the particular form of effect, be i.e. obligate pathogenetic bacteria, facultative spore bacteria, facultative pathogenetic bacteria and the pathogenetic bacteria that hides.
(1) obligate specialized hospital bacterium obligate pathogenetic bacteria is the extremely strong double anaerobies bacillus of a class obligate, has very strong pathogenicity to specific insect host.As chafer emulsus disease pathogen bacterium, it is class obligate bacillus chafer larva being had to pathogenicity.According to reporting for work, countries in the world have isolated emulsus disease pathogen bacterium from different regions multiple, wherein important with the dead bacillus Australia mutation of easing up of Bacillus popilliae, slow dead bacillus, they can produce pathogenicity (Dutky, 1963) to 23 kinds of chafer larvas.
(2) the facultative spore bacteria of facultative spore bacteria refer to can produce gemma and can on synthetic medium normal growth grow a bacterioid, wherein have the spore bacteria bacillus thuringiensis,Bt monoid that can produce parasporal crystal and the bacillus not producing crystal.Wax bacillus is the representative species of the bacillus not producing crystal, be a kind of bacillus often can found in soil, and separate in the Chang Kecong insect bodies such as coleoptera, Lepidoptera, Hymenoptera of falling ill, infected larva is stiff, body wall is black dull, and polypide is softening to be shunk.Larva through injection treatment is often dead in 14 ~ 48h.
(3) that hides lives in the digestive tract of insect usually without this bacterioid of gemma pathogenetic bacteria, mainly some asporogenic bacillus, comprise the gas bacillus, proteus etc. of the verdigris pole hair bacillus of pseudomonas section, pseudomonas fluorescens and enterobacteriaceae.Seldom cause disease at ordinary times, once enter insect body cavity, then can show stronger pathogenicity.The ability that these bacteriums enter body cavity by middle intestines depends mainly on some incitant, cause morbidity as extreme humidity, insect eat the wound etc. that strange food, the infecting of other cause of diseases, parasite or cannibalism cause, be generally common in the insect population of artificial feeding.
(4) facultative without gemma pathogenetic bacteria as clayey Sai Shi bacillus, can have tens of kinds by parasitic insect, be Gram negative bacillus pumilis, do not produce gemma, bacterium colony be pale red or cerise on agar medium, is easy to identification, also adularescent bacterial strain.Under field conditions (factors), clayey Sai Shi bacillus can be found in corn borer, black cutworm, corn earworm wait for death polypide.Although this bacterium also has potency feature, cause disease comparatively common, there is certain ability invading haemocoele, therefore be classified as facultative cause of disease.After this bacterium invades polypide haemocoele, septicemia can be caused in 1 ~ 3d to make host dead, with the most common in the Natural Population of locust section.In the disease of insect, infect by disease fungus the mycosis (mycosis) caused and account for 60%.Fungi is the eucaryote of the low grade of a class, unicellular, or the cell broken up by majority formed, and not containing chlorophyll, can not seek photosynthesis and Formed hydrate, without the organ such as root, stem, leaf of differentiation.The trophosome of fungi is mycelia, and most of fungi can breed with mycelia fragment or mycelia fracture, and most fungi again can with sporogenesis.Larger than bacterium several times to tens times of the fungi relevant with insect, hypha,hyphae mean breadth is 3 ~ 10 μm.Mycelia is disagreed, some without tabula, whole mycelia be one unicellular, as Phycomycetes; What have has tabula, is many cells, as Ascomycetes and deuteromycetes etc.
The modes of reproduction of disease fungus can be divided into vegetative propagation and sexual propagation two kinds.Vegetative propagation forms asexual spore, is divided into again sporangiospore and conidium two class.Most fungus produces spores cystospore without tabula, breaks after sporangium maturation, spreads out spore, spore sprouts, grows, forms new individuality.Conidiophore can be carried and wide dispersion by wind, water and animal, is that disease fungus infects the topmost approach of insect.Sexual propagation is most in the fungi of mycelia without tabula produces zygospore, and the fungi majority with tabula produces ascospore.Most fungi has asexual and sexual two eposides in the history of life; often alternate with each other, the alternation of generations of meaning, the bacterium colony of the bacterium of fungi is usually in fine hair shape or loose flocculence; its nutrient hypha and aerial hyphae all can form different pigments, and spore also has different colors.Some fungi of infecting insect is obligatory parasitic bacteria, relies on life host for its nutritional need.There are some to be half parasite, also have general saprophytic bacteria.Saprophytic growing also can cause susceptible insect that obvious disease occurs under certain conditions, but the secondary natural disposition saprophytic bacteria after polypide death, body grown easily is mistaken as lethal germ.
The present invention has the following advantages:
1, the microorganism that the present invention selects will possess more following basic attributes:
1. virulence is high, and microbial inoculum virulence is enough to the premunition overwhelming insect, can reliablely and stablely make insect fall ill lethal.
2. good stability, microbial inoculum not reason lowers insecticidal power by impacts such as natural environment (as daylight, drying etc.), artificial treatment, suspending agent, carrier, storage, different insecticide-applying way.
3. the longevity of residure is long, and has the ability of natural propagation in pest population.
4. effect rapidly.
5. selective, namely to controlling object severe toxicity, and nontoxic to plant, beneficial insect and mammal etc.
6. can large-scale production, economic security.
2, preparation method of the present invention will have the different bacterial classification of typical three class 18 kinds above, first by the sandy soil pipe bacterial classification bought back under freezen protective of National agricultural bacterial classification Stock Center, cultivates through test tube slant, laboratory; Sediments microscope inspection confirms; Triangular flask shaking table is cultivated; Seed fermentation tank; Produce fermentation tank; Hold-up tank produce liquid product; Add filler; Filter; Filter cake; To size mixing tank; Spray business is dry; Obtain solid bacterium powder; Laboratory quality inspection; Packed products; Warehouse-in.
3, essential core of the present invention is the insecticide for general microorganism in the market, and desinsection object is single, the feature that insecticidal power is weak.Programmed screening 18 kinds kills the strong bacterial classification of kind of effect and forms powerful cenobium, selects three class bacterial classifications get the diversified broad spectrum activity feature of its desinsection and build.
Embodiment
Embodiment 1
Prepare antimicrobial composition insecticide (units/kg):
The first step: Spawn incubation: according to title and the numbering of bacterial classification, buys three major types 18 sand pipe bacterial classifications back by China Microbiological preservation committee agricultural microorganism center; Three quasi-microorganisms are fungi, actinomycetes and bacterium.
Fungi has 3, be respectively: stiff bacterium ACCC-30101 ← clay fertilizer institute of the Chinese Academy of Agricultural Sciences ← sand control institute of AS3.3675 ← Shan Xi Yu forest land district of Institute of Microorganism, Academia Sinica of (1) metarhiziumaunisopliae (metschn) sorok chafer edge, is separated from paranthrene tabaniformis larva; For anti-corn borer, chafer, clearwing moth etc.(2) Paecilomycesfarinosus (Dicks.) Brown turns ceramic route ACCC-30190 ← Chinese Academy of Agricultural Sciences's clay fertilizer institute ← Beijing Forestry University's force literary composition of presenting oneself before (a monarch) is given; Original number: 8601, control dendrolimus tabulaeformis.(3) bright giving is wished by Paecilomyceslilacinus (Thom) Samson paecilomyces lilacinus ACCC-30677 ← tobacco research institute of Yunnan Province; Original number: ZML086; Locality: Yunnan; Separation source: root-knot nematode egg; Study as biological control.
4, actinomycetes, are respectively: (1) Streptomycesanulatus (Beijingrinck1912) wdksman1953 garden ring streptomycete ACCC-41456 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00797; Metabolite has insecticidal activity; Separation source: soil; Be separated place: Lushi, henan five li of rivers.(2) raw institute is planted in " Streptomycesculicidicus " Yanetal. streptomyces culicidicus ACCC-41132 ← institute of microbiology ← Shanghai, Guangzhou, Guangdong Province; Original number: GIM4.12MIK4.1; Kill mosquito larvae; Separation source: soil; Be separated place: Guangzhou.(3) StreptomycesgriseolussubsphangzhouensisACCC-40075 ← Zhejiang Agriculture academy of sciences; Original number: S26; Product kills aphid element, very strong to aphid, two spotted spider mite and some lepidopterous larvae toxic action.(4) StreptomycescalvusBackusetal., 1957 bald naked streptomycete ACCC-41451 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00743; Metabolite has insecticidal activity; Separation source: soil; Be separated place: Zhejiang dragon trip.
11, bacterium, is respectively: (1) Bacillussubtilis (Enrenberg1835) cohn1872 bacillus subtilis ACCC-11088 ← Institute of Microorganism, Academia Sinica; AS1.0933; Control peanut nematodosis.(2) BacillusSphaericusmeyerandNeide1904 Bacillus sphaericus ACCC-11096 ← Institute of Microorganism, Academia Sinica ← U.S.;=AS1.1672; Original number: 2115; Kill mosquito.(3) BacilluslaterosporusLaubach bacillus laterosporus ACCC-11079 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute ← Microbe Inst., Chinese Academy of Sciences AS1.0864; Larvae can be killed; Produce lysozyme.(4) Bacillusthuringienissubsp.AisawaiACCC-10016 ← Wuhan microbial pesticide factory; Original number: 096; Prepare antigen reference culture, poisoning lepidopterous insects.(5) Bacillusthuringienissubsp.merrisoniACCC-10031 ← Wuhan microbial pesticide factory; Original number: 012; Poisoning lepidopterous insects.(6) Bacillusthuringiensissubsp.galleriae bacillus thuringiensis,Bt galleria mellonella waxmoth subspecies ACCC-10027 ← Microbe Inst., Chinese Academy of Sciences; Poisoning cabbage caterpillar.(7) Bacillusthuringiensissubsp.Israelensis Bacillus thuringiensis subsp. israelensis ACCC-10038 ← Chengdu Hua Hong industrial corporation ← Taian Shandong biological study institute ← World Health Organization (WHO); Kill mosquitoes larva, Jue
effect is better.(8) Bacillusthuringienissubsp.thuringienisACCC-10035 ← China Agriculture Academe Fertilizer Institute; Original number: D25; Antiphagin poisoning corn borer; Separation source: spontaneons screening from HD-187 bacterial strain and obtaining.(9) Bacillusthuringiensissuhsp.berliner bacillus thuringiensis,Bt Bai Shi mutation ACCC-10022 ← China Agriculture Academe Fertilizer Institute; Original number: D26; Control lepidopterous insects; Separation source: bacterium powder.(10) Bacillusthuringiensissubsp.Dendrolimus bacillus thuringiensis,Bt pine another name for Sichuan Province subspecies ACCC-10062 ← Soviet Union; Poisoning pine caterpillars.(11) Bacillusthuringienissubsp.sottoACCC-10033 ← Wuhan microbial pesticide factory ← Britain; Original number: 016; Prepare antigen reference culture, poisoning lepidopterous insects.
Aseptically these sand pipe bacterial classifications are moved respectively in the slant tube received and beef extract, peptone agar medium are housed; Observe its purity, without plaque, have more than 95% form normal without miscellaneous bacteria, smear for microscopic examination; Normal bacterial classification good for purity is carried out single thalline liquid shaking table to cultivate; Quantity expands shaking table facility scale or repeats to purchase once on request, meets the requirement that next step is cultivated.
Second step: the expansion of bacterium liquid is cultivated: first mix similar for 18 bacterium liquid in the ratio of 1: 1 respectively, namely fungi, actinomycetes, bacterium class obtain pure class bacterium mixed liquor respectively; Mix by the weight ratio of this three major types type mixed liquor fungi 50, actinomycetes 20, bacterium 30 again; Mixed bacteria liquid 10kg is added 90kg in purification wild Oryza species liquid, liquid medium ingredients by weight part ratio is analysis for soybean powder 1.2, dextrin 0.6, peptone 0.3, dipotassium hydrogen phosphate 0.2, calcium carbonate 0.5, magnesium sulfate 0.4, ammonium sulfate 0.05, soya-bean oil 0.2, pH value be 7.2 join in nutrient solution; Put into seeding tank, at temperature 30 DEG C ± 1 DEG C, stirring per hour 200 times, cultivates 44 hours; Microscopy every gram bacteria containing amount reaches 2,000,000,000/gram time, stop cultivating, check and be registered seed bacterium liquid without miscellaneous bacteria.By qualified bacterium liquid in the ratio of 5%, add in fermentation tank, in fermentation tank, liquid medium as described above; Cultivate 38 hours, on inspection without miscellaneous bacteria, bacterium liquid obviously becomes gemma and crystal, and content reaches 2,000,000,000/gram, namely can be finished product fermentation product.
3rd step: prepare solid fungicide further: first get above-mentioned finished product fermentation product bacterium liquid, add the calcium carbonate of 9% by weight, stir 30 minutes; Pass through plate-frame filtering; Stir 30 minutes, furnishing pulpous state; Stamp head tank again, carry out spraying dry; Bacterium powder after spraying dry, through being pulverized and mixed evenly, is taken a sample to check qualified; Timely plastic bag packaging seals; Put into shady and cool dry storehouse to preserve.
Embodiment 2
Prepare antimicrobial composition insecticide (units/kg):
The first step: Spawn incubation: according to title and the numbering of bacterial classification, buys fungi, actinomycetes, bacteria culture back by China Microbiological preservation committee agricultural microorganism center;
Fungi 2: the stiff bacterium ACCC of (1) metarhiziumaunisopliae (metschn) sorok chafer edge---clay fertilizer institute of 30101 ← Chinese Academy of Agricultural Sciences ← sand control institute of AS3.3675 ← Shan Xi Yu forest land district of Institute of Microorganism, Academia Sinica, is separated from paranthrene tabaniformis larva; For anti-corn borer, chafer, clearwing moth etc.
(2) Paecilomycesfarinosus (Dicks.) Brown turns ceramic route ACCC---30190 ← Chinese Academy of Agricultural Sciences clay fertilizer institute ← Beijing Forestry University's force literary composition of presenting oneself before (a monarch) is given; Original number: 8601, control dendrolimus tabulaeformis.
3, actinomycetes, are respectively: (1) Streptomycesanulatus (Beijingrinck1912) wdksman1953 garden ring strepto-ACCC-41456 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00797; Metabolite has insecticidal activity; Separation source: soil; Be separated place: Lushi, henan five li of rivers.(2) raw institute is planted in " Streptomycesculicidicus " Yanetal. streptomyces culicidicus ACCC-41132 ← institute of microbiology ← Shanghai, Guangzhou, Guangdong Province; Original number: GIM4.12MIK4.1; Kill mosquito larvae; Separation source: soil; Be separated place: Guangzhou.(3) StreptomycesgriseolussubsphangzhouensisACCC-40075 ← Zhejiang Agriculture academy of sciences; Original number: S26; Product kills aphid element, very strong to aphid, two spotted spider mite and some lepidopterous larvae toxic action.
9, bacterium, is respectively: (1) Bacillussubtilis (Enrenberg1835) cohn1872 bacillus subtilis ACCC-11088 ← Institute of Microorganism, Academia Sinica; AS1.0933; (2) BacillusSphaericusmeyerandNeide1904 Bacillus sphaericus ACCC-11096 ← Institute of Microorganism, Academia Sinica ← U.S.;=AS1.1672; Original number: 2115; (3) BacilluslaterosporusLaubach bacillus laterosporus ACCC-11079 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute ← Microbe Inst., Chinese Academy of Sciences AS1.0864; (4) Bacillusthuringienissubsp.AisawaiACCC-10016 ← Wuhan microbial pesticide factory; Original number: 096; (5) Bacillusthuringienissubsp.merrisoniACCC-10031 ← Wuhan microbial pesticide factory; Original number: 012; (6) Bacillusthuringiensissubsp.galleriae bacillus thuringiensis,Bt galleria mellonella waxmoth subspecies ACCC-10027 ← Microbe Inst., Chinese Academy of Sciences; (7) Bacillusthuringiensissubsp.Israelensis Bacillus thuringiensis subsp. israelensis ACCC-10038 ← Chengdu Hua Hong industrial corporation ← Taian Shandong biological study institute ← World Health Organization (WHO); (8) Bacillusthuringienissubsp.thuringienisACCC-10035 ← China Agriculture Academe Fertilizer Institute; Original number: D25; (9) Bacillusthuringiensissuhsp.berliner bacillus thuringiensis,Bt Bai Shi mutation ACCC-10022 ← China Agriculture Academe Fertilizer Institute; Original number?
Aseptically these sand pipe bacterial classifications are moved respectively in the slant tube received and beef extract, peptone agar medium are housed; Observe its purity, without plaque, have more than 95% form normal without miscellaneous bacteria, smear for microscopic examination; Normal bacterial classification good for purity is carried out single thalline liquid shaking table to cultivate; Quantity expands shaking table facility scale or repeats to purchase once on request, meets the requirement that next step is cultivated.
Second step: the expansion of bacterium liquid is cultivated: first divided by bacterium liquid three similarly to mix in the ratio of 1: 1 respectively, i.e. fungi, actinomycetes, bacterium obtained similar bacterium mixed liquor respectively; Mix by this mixed liquor fungi 40 of the same type, actinomycetes 20, bacterium 40 weight ratio again; Mixed bacteria liquid 10kg is added 80kg in purification wild Oryza species liquid, liquid medium ingredients by weight part ratio is analysis for soybean powder 1, dextrin 0.3, peptone 0.2, dipotassium hydrogen phosphate 0.1, calcium carbonate 0.3, magnesium sulfate 0.2, ammonium sulfate 0.03, soya-bean oil 0.1, pH value be 7.2 join in nutrient solution; Put into seeding tank, at temperature 28 DEG C, stirring per hour 150 times, cultivates 36 hours; Microscopy every gram bacteria containing amount reaches 2,000,000,000/gram time, stop cultivating, check and be registered seed bacterium liquid without miscellaneous bacteria.By qualified bacterium liquid in the ratio of 3%, add in fermentation tank, in fermentation tank, liquid medium as described above; Cultivate 30 hours, on inspection without miscellaneous bacteria, bacterium liquid obviously becomes gemma and crystal, and content reaches 2,000,000,000/gram, namely can be finished product fermentation product.
3rd step: prepare solid fungicide further: first get above-mentioned finished product fermentation product bacterium liquid, add the calcium carbonate of 8%k by weight, stir 30 minutes; Pass through plate-frame filtering; Stir 30 minutes, furnishing pulpous state; Stamp head tank again, carry out spraying dry; Bacterium powder after spraying dry, through being pulverized and mixed evenly, is taken a sample to check qualified; Timely plastic bag packaging seals; Put into shady and cool dry storehouse to preserve.
Embodiment 3
Prepare antimicrobial composition insecticide (units/kg):
The first step: Spawn incubation: according to title and the numbering of bacterial classification, buys fungi, actinomycetes sand pipe bacterial classification back by China Microbiological preservation committee agricultural microorganism center;
Fungi has 3, be respectively: the stiff bacterium ACCC of (1) metarhiziumaunisopliae (metschn) sorok chafer edge---clay fertilizer institute of 30101 ← Chinese Academy of Agricultural Sciences ← sand control institute of AS3.3675 ← Shan Xi Yu forest land district of Institute of Microorganism, Academia Sinica, is separated from paranthrene tabaniformis larva; For anti-corn borer, chafer, clearwing moth etc.
(2) Paecilomycesfarinosus (Dicks.) Brown turns ceramic route ACCC---30190 ← Chinese Academy of Agricultural Sciences clay fertilizer institute ← Beijing Forestry University's force literary composition of presenting oneself before (a monarch) is given; Original number: 8601, control dendrolimus tabulaeformis.(3) bright giving is wished by ACCC---tobacco research institute of 30677 ← Yunnan Province; Original number: ZML086; Locality: Yunnan; Separation source: root-knot nematode egg; Study as biological control.
2, actinomycetes, are respectively: Streptomycesgriseolussubsphangzhouensis
ACCC---40075 ← Zhejiang Agriculture academy of sciences; Original number: S26; Product kills aphid element, very strong to aphid, two spotted spider mite and some lepidopterous larvae toxic action.StreptomycescalVusBackusetal., 1957 bald naked streptomycete ACCC---41451 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00743; Metabolite has insecticidal activity; Separation source: soil; Be separated place: Zhejiang dragon trip.
Aseptically these sand pipe bacterial classifications are moved respectively in the slant tube received and beef extract, peptone agar medium are housed; Observe its purity, without plaque, have more than 95% form normal without miscellaneous bacteria, smear for microscopic examination; Normal bacterial classification good for purity is carried out single thalline liquid shaking table to cultivate; Quantity expands shaking table facility scale or repeats to purchase once on request, meets the requirement that next step is cultivated.
Second step: the expansion of bacterium liquid is cultivated: first mix similar for bacterium liquid in the ratio of 0.5: 1.5 respectively, i.e. fungi, actinomycetes obtained similar bacterium mixed liquor respectively; Mix by the weight ratio of this mixed liquor fungi 60, actinomycetes 20 again; Mixed bacteria liquid 10kg is added 100kg in purification wild Oryza species liquid, liquid medium ingredients by weight part ratio is analysis for soybean powder 1.5, dextrin 0.9, peptone 0.4, dipotassium hydrogen phosphate 0.3, calcium carbonate 0.7, magnesium sulfate 0.6, ammonium sulfate 0.07, soya-bean oil 0.3, pH value be 7.2 join in nutrient solution; Put into seeding tank, at temperature 30 DEG C, stirring per hour 200 times, cultivates 48 hours; Microscopy every gram bacteria containing amount reaches 2,000,000,000/gram time, stop cultivating, check and be registered seed bacterium liquid without miscellaneous bacteria.By qualified bacterium liquid in the ratio of 6%, add in fermentation tank, in fermentation tank, liquid medium as described above; Cultivate 45 hours, on inspection without miscellaneous bacteria, bacterium liquid obviously becomes gemma and crystal, and content reaches 2,000,000,000/gram, namely can be finished product fermentation product.
3rd step: prepare solid fungicide further: first get the calcium carbonate that above-mentioned finished product fermentation product bacterium liquid adds 10% by weight, stir 30 minutes; Pass through plate-frame filtering; Stir 30 minutes, furnishing pulpous state; Stamp head tank again, carry out spraying dry; Bacterium powder after spraying dry, through being pulverized and mixed evenly, is taken a sample to check qualified; Timely plastic bag packaging seals; Put into shady and cool dry storehouse to preserve.
Embodiment 4
Prepare antimicrobial composition insecticide (units/kg):
The first step: Spawn incubation: according to title and the numbering of bacterial classification, buys three major types sand pipe bacterial classification back by China Microbiological preservation committee agricultural microorganism center; Three quasi-microorganisms are fungi, actinomycetes and bacterium.
Fungi has 2, is respectively: Paecilomycesfarinosus (Dicks.) Brown turns ceramic route ACCC-30190 ← Chinese Academy of Agricultural Sciences's clay fertilizer institute ← Beijing Forestry University's force literary composition of presenting oneself before (a monarch) is given; Original number: 8601, control dendrolimus tabulaeformis.Bright giving is wished by Paecilomyceslilacinus (Thom) Samson paecilomyces lilacinus ACCC-30677 ← tobacco research institute of Yunnan Province; Original number: ZML086; Locality: Yunnan; Separation source: root-knot nematode egg; Study as biological control.
4, actinomycetes, are respectively: (1) Streptomycesanulatus (Beijingrinck1912) wdksman1953 garden ring streptomycete ACCC-41456 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00797; Metabolite has insecticidal activity; Separation source: soil; Be separated place: Lushi, henan five li of rivers.(2) raw institute is planted in " Streptomycesculicidicus " Yanetal. kill mosquito strepto-ACCC-41132 ← institute of microbiology ← Shanghai, Guangzhou, Guangdong Province; Original number: GIM4.12MIK4.1; Kill mosquito larvae; Separation source: soil; Be separated place: Guangzhou.(3) StreptomycesgriseolussubsphangzhouensisACCC-40075 ← Zhejiang Agriculture academy of sciences; Original number: S26; Product kills aphid element, very strong to aphid, two spotted spider mite and some lepidopterous larvae toxic action.(4) StreptomycescalvusBackusetal., 1957 bald naked streptomycete ACCC-41451 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00743; Metabolite has insecticidal activity; Separation source: soil; Be separated place: Zhejiang dragon trip.
8, bacterium, is respectively: (1) Bacillussubtilis (Enrenberg1835) cohn1872 bacillus subtilis ACCC-11088 ← Institute of Microorganism, Academia Sinica; AS1.0933; (2) BacillusSphaericusmeyerandNeide1904 Bacillus sphaericus ACCC-11096 ← Institute of Microorganism, Academia Sinica ← U.S.;=AS1.1672; Original number: 2115; (3) BacilluslaterosporusLaubach bacillus laterosporus ACCC-11079 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute ← Microbe Inst., Chinese Academy of Sciences AS1.0864; (4) Bacillusthuringienissubsp.AisawaiACCC-10016 ← Wuhan microbial pesticide factory; Original number: 096; (5) Bacillusthuringienissubsp.merrisoniACCC-10031 ← Wuhan microbial pesticide factory; Original number: 012; (6) Bacillusthuringiensissubsp.galleriae bacillus thuringiensis,Bt galleria mellonella waxmoth subspecies ACCC-10027 ← Microbe Inst., Chinese Academy of Sciences; (7) Bacillusthuringiensissubsp.Israelensis Bacillus thuringiensis subsp. israelensis ACCC-10038 ← Chengdu Hua Hong industrial corporation ← Taian Shandong biological study institute ← World Health Organization (WHO); (8) Bacillusthuringienissubsp.thuringienisACCC-10035 ← China Agriculture Academe Fertilizer Institute; Original number: D25;
Aseptically these sand pipe bacterial classifications are moved respectively in the slant tube received and beef extract, peptone agar medium are housed; Observe its purity, without plaque, have more than 95% form normal without miscellaneous bacteria, smear for microscopic examination; Normal bacterial classification good for purity is carried out single thalline liquid shaking table to cultivate; Quantity expands shaking table facility scale or repeats to purchase once on request, meets the requirement that next step is cultivated.
Second step: the expansion of bacterium liquid is cultivated: first mix similar for bacterium liquid in the ratio of 1: 1 respectively, i.e. fungi, actinomycetes, bacterium class obtained similar bacterium mixed liquor respectively; Mix by the weight ratio of this three major types type mixed liquor fungi 60, actinomycetes 25, bacterium 40 again. mixed bacteria liquid 10kg is added 90kg in purification wild Oryza species liquid, liquid medium ingredients by weight part ratio is analysis for soybean powder 1.2, dextrin 0.6, peptone 0.3, dipotassium hydrogen phosphate 0.2, calcium carbonate 0.5, magnesium sulfate 0.4, ammonium sulfate 0.05, soya-bean oil 0.2, pH value be 7.2 join in nutrient solution; Put into seeding tank, at temperature 30 DEG C ± 1 DEG C, stirring per hour 200 times, cultivates 36 hours; Microscopy every gram bacteria containing amount reaches 2,000,000,000/gram time, stop cultivating, check and be registered seed bacterium liquid without miscellaneous bacteria.By qualified bacterium liquid in the ratio of 5%, add in fermentation tank, in fermentation tank, liquid medium as described above; Cultivate 45 hours, on inspection without miscellaneous bacteria, bacterium liquid obviously becomes gemma and crystal, and content reaches 2,000,000,000/gram, namely can be finished product fermentation product.
3rd step: prepare solid fungicide further: first get above-mentioned finished product fermentation product bacterium liquid, add the calcium carbonate of 9% by weight, stir 30 minutes; Pass through plate-frame filtering; Stir 30 minutes, furnishing pulpous state; Stamp head tank again, carry out spraying dry; Bacterium powder after spraying dry, through being pulverized and mixed evenly, is taken a sample to check qualified; Timely plastic bag packaging seals; Put into shady and cool dry storehouse to preserve.Fungi 40-60, actinomycetes 15-25 and/or bacterium 20-40;
Embodiment 5
Prepare antimicrobial composition insecticide (units/kg):
The first step: Spawn incubation: according to title and the numbering of bacterial classification, buys actinomycetes and bacteria culture back by China Microbiological preservation committee agricultural microorganism center;
3, actinomycetes, are respectively: raw institute is planted in (1) " Streptomycesculicidicus " Yanetal. streptomyces culicidicus ACCC-41132 ← institute of microbiology ← Shanghai, Guangzhou, Guangdong Province; Original number: GIM4.12MIK4.1; Kill mosquito larvae; Separation source: soil; Be separated place: Guangzhou.(2) StreptomycesgriseolussubsphangzhouensisACCC-40075 ← Zhejiang Agriculture academy of sciences; Original number: S26; Product kills aphid element, very strong to aphid, two spotted spider mite and some lepidopterous larvae toxic action.(3) StreptomycescalVusBackusetal., 1957 bald naked streptomycete ACCC-41451 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00743; Metabolite has insecticidal activity; Separation source: soil; Be separated place: Zhejiang dragon trip.
6, bacterium, is respectively: (1) Bacillusthuringienissubsp.merrisoniACCC---10031 ← Wuhan microbial pesticide factory; Original number: 012; Poisoning lepidopterous insects.(2) Bacillusthuringiensissubsp.galleriae bacillus thuringiensis,Bt galleria mellonella waxmoth subspecies ACCC-10027 ← Microbe Inst., Chinese Academy of Sciences; Poisoning cabbage caterpillar.(3) Bacillusthuringiensissubsp.Israelensis Bacillus thuringiensis subsp. israelensis ACCC-10038 ← Chengdu Hua Hong industrial corporation ← Taian Shandong biological study institute ← World Health Organization (WHO); Kill mosquitoes larva, Jue
effect is better.(4) Bacillusthuringienissubsp.thuringienisACCC-10035 ← China Agriculture Academe Fertilizer Institute; Original number: D25; Antiphagin poisoning corn borer; Separation source: spontaneons screening from HD-187 bacterial strain and obtaining.(5) Bacillusthuringiensissuhsp.berliner bacillus thuringiensis,Bt Bai Shi mutation ACCC-10022 ← China Agriculture Academe Fertilizer Institute; Original number: D26; Control lepidopterous insects; Separation source: bacterium powder.(6) Bacillusthuringiensissubsp.Dendrolimus bacillus thuringiensis,Bt pine another name for Sichuan Province subspecies ACCC-10062 ← Soviet Union; Poisoning pine caterpillars.
Preparation method is with embodiment 1.Difference is: second step: the expansion of bacterium liquid is cultivated: first mixed in the ratio of 1: 1 respectively by similar for bacterial classification pure liquid, i.e. actinomycetes, bacterium class obtained mixed liquor respectively; Mix by the weight ratio of these three major types type mixed liquor actinomycetes 20, bacterium 30 again.
Embodiment 6
Prepare antimicrobial composition insecticide (units/kg):
The first step: Spawn incubation: according to title and the numbering of bacterial classification, buys fungi, bacteria culture back by China Microbiological preservation committee agricultural microorganism center;
Fungi has 3, be respectively: the stiff bacterium ACCC of (1) metarhiziumaunisopliae (metschn) sorok chafer edge---clay fertilizer institute of 30101 ← Chinese Academy of Agricultural Sciences ← sand control institute of AS3.3675 ← Shan Xi Yu forest land district of Institute of Microorganism, Academia Sinica, is separated from paranthrene tabaniformis larva; For anti-corn borer, chafer, clearwing moth etc.
(2) Paecilomycesfarinosus (Dicks.) Brown turns ceramic route ACCC---30190 ← Chinese Academy of Agricultural Sciences clay fertilizer institute ← Beijing Forestry University's force literary composition of presenting oneself before (a monarch) is given; Original number: 8601, control dendrolimus tabulaeformis.(3) bright giving is wished by Paecilomyceslilacinus (Thom) Samson paecilomyces lilacinus ACCC---tobacco research institute of 30677 ← Yunnan Province; Original number: ZML086; Locality: Yunnan; Separation source: root-knot nematode egg; Study as biological control.
11, bacterium, is respectively: (1) Bacillussubtilis (Enrenberg1835) cohn1872 bacillus subtilis ACCC-11088 ← Institute of Microorganism, Academia Sinica; AS1.0933; Control peanut nematodosis.(2) BacillusSphaericusmeyerandNeide1904 Bacillus sphaericus ACCC-11096 ← Institute of Microorganism, Academia Sinica ← U.S.;=AS1.1672; Original number: 2115; Kill mosquito.(3) BacilluslaterosporusLaubach bacillus laterosporus ACCC-11079 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute ← Microbe Inst., Chinese Academy of Sciences AS1.0864; Larvae can be killed; Produce lysozyme.(4) Bacillusthuringienissubsp.AisawaiACCC-10016 ← Wuhan microbial pesticide factory; Original number: 096; Prepare antigen reference culture, poisoning lepidopterous insects.(5) Bacillusthuringienissubsp.merrisoniACCC-10031 ← Wuhan microbial pesticide factory; Original number: 012; Poisoning lepidopterous insects.(6) Bacillusthuringiensissubsp.galleriae bacillus thuringiensis,Bt galleria mellonella waxmoth subspecies ACCC-10027 ← Microbe Inst., Chinese Academy of Sciences; Poisoning cabbage caterpillar.(7) Bacillusthuringiensissubsp.Israelensis Bacillus thuringiensis subsp. israelensis ACCC-10038 ← Chengdu Hua Hong industrial corporation ← Taian Shandong biological study institute ← World Health Organization (WHO); Kill mosquitoes larva, Jue
effect is better.(8) Bacillusthuringienissubsp.thuringienisACCC-10035 ← China Agriculture Academe Fertilizer Institute; Original number: D25; Antiphagin poisoning corn borer; Separation source: spontaneons screening from HD-187 bacterial strain and obtaining.(9) Bacillusthuringiensissuhsp.berliner bacillus thuringiensis,Bt Bai Shi mutation ACCC-10022 ← China Agriculture Academe Fertilizer Institute; Original number: D26; Control lepidopterous insects; Separation source: bacterium powder.(10) Bacillusthuringiensissubsp.Dendrolimus bacillus thuringiensis,Bt pine another name for Sichuan Province subspecies ACCC-10062 ← Soviet Union; Poisoning pine caterpillars.(11) Bacillusthuringienissubsp.sottoACCC-10033 ← Wuhan microbial pesticide factory ← Britain; Original number: 016; Prepare antigen reference culture, poisoning lepidopterous insects.
Preparation method is with embodiment 1, and difference is second step: the expansion of bacterium liquid is cultivated: first mixed in the ratio of 1: 1 respectively by similar for bacterial classification pure liquid, i.e. fungi, bacterium class obtained mixed liquor respectively; Mix by the weight ratio of this three major types type mixed liquor fungi 50, bacterium 30 again.
Embodiment 7
Embodiment 1 finished product of preparation is sprayed on pest body epidermis by the present invention, the imbibition of bacterium spore, about can sprout through 8-12 hour and grow the germination of 1-2 bar, secretion chitin lyase, invade to sprout germ tube, later elongation is mycelia, directly absorbs mixed polypide liquid nutrient and arrives and fill the air in polypide blood, hindering blood circulation.The metabolite Oxalates of mycelia accumulates in blood in a large number, and basicity declines, and loses original transparency and stagnant muddiness, causes the change of physicochemical property, makes metabolic disorders and dead.
Embodiment 8
By Finished Product Examples 2 preparation of the present invention, be sprayed on plant, enter enteron aisle through insect population, within 48 hours, enteron aisle miospore germination rate is 37%.In 60-72 hour, mycelia invades intestinal walls, and makes digestive juice enter haemocoele, and within 84 hours, digestive juice pH value is down to 5.0 by 7.0, and mycelia stretches out from polypide intersegmental membrane, valve, sense organ duct, and lethality reaches 95%.
Embodiment 9
By Finished Product Examples 3 preparation of the present invention, be sprinkled upon on polypide, fungi is poisonous to insect, but directly do not kill insect, and destroy polypide epidermis, be conducive to insecticidal bacteria and invade, cause syndrome, do not treat the mycotic slow generating process of onset, and dead rapidly by bacterium system disease.Just prove that the present invention utilizes the correct and preparation as insecticiding Be very effective of the innovative idea of multiclass bacterium mixed insecticidal.
Embodiment 10
Antimicrobial composition embodiment 6 insecticide of the present invention and market single culture insecticide are to a variety of Chinese cabbage field cabbage caterpillar poison test.Spray insecticide of the present invention after five days, group 32 cabbage caterpillars in test region have 25 to occur pathology or death; Group 31 cabbage caterpillars in check plot have 12 pathology or death occur.Prove that this insecticide is higher by 51% than contrast insecticidal effect.
Claims (10)
1. an antimicrobial composition insecticide is by the concentrated cenobium preparation being selected from fungi, 2 classes of actinomycetes and/or bacterium or 3 class bacterial classifications and being prepared into; Described parts by weight of component ratio is: fungi 40-60, actinomycetes 15-25 and/or bacterium 20-40.
2. antimicrobial composition insecticide according to claim 1, described parts by weight of component ratio is: fungi 50, actinomycetes 20 and/or bacterium 30.
3., according to the antimicrobial composition insecticide of claim 1 or 2, described fungi, is selected from following 2 kinds or two or more bacterial classification:
(1) stiff bacterium ACCC-30101 ← clay fertilizer institute of the Chinese Academy of Agricultural Sciences ← sand control institute of AS3.3675 ← Shan Xi Yu forest land district of Institute of Microorganism, Academia Sinica of metarhiziumanisopliae (metschn) sorok chafer edge;
(2) Paecilomycesfarinosus (Dicks.) Brown turns ceramic route ACCC-30190 ← Chinese Academy of Agricultural Sciences's clay fertilizer institute ← Beijing Forestry University's force literary composition of presenting oneself before (a monarch) is given, original number: 8601;
(3) bright giving is wished by Paecilomyceslilacinus (Thom) Samson paecilomyces lilacinus ACCC-30677 ← tobacco research institute of Yunnan Province; Original number: ZML086;
Described actinomycetes, are selected from following 2 kinds or two or more bacterial classification:
(1) Streptomycesanuiatus (Beijingrinck1912) wadksman1953 garden ring streptomycete ACCC-41456 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00797;
(2) raw institute is planted in " Streptomycesculicidicus " Yanetal. streptomyces culicidicus ACCC-41132 ← institute of microbiology ← Shanghai, Guangzhou, Guangdong Province; Original number: GIM4.12MIK4.1;
(3) StreptomycesgriseolussubsphangzhouensisACCC-40075 ← Zhejiang Agriculture academy of sciences; Original number: S26;
(4) StreptomycescaiVusBackusetal., 1957 bald naked streptomycete ACCC-41451 ← Hubei Province biological pesticide Engineering Research Center; Original number: HBERC00743;
Described bacterium, is selected from following 2 kinds or two or more bacterial classification:
(1) Bacillussubtilis (Enrenberg1835) cohn1872 bacillus subtilis ACCC-11088 ← Institute of Microorganism, Academia Sinica; AS1.0933;
(2) BacillusSphaericusmeyerandNeide1904 Bacillus sphaericus ACCC-11096 ← Institute of Microorganism, Academia Sinica ← U.S.;=AS1.1672; Original number: 2115;
(3) BacilluslaterosporusLaubach bacillus laterosporus ACCC-11079 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute ← Microbe Inst., Chinese Academy of Sciences AS1.0864;
(4) Bacillusthuringienissubsp.AisawaiACCC-10016 ← Wuhan microbial pesticide factory; Original number: 096;
(5) Bacillusthuringienissubsp.merrisoniACCC-10031 ← Wuhan microbial pesticide factory; Original number: 012;
(6) Bacillusthuringiensissusp.galleriae bacillus thuringiensis,Bt galleria mellonella waxmoth subspecies ACCC-10027 ← Microbe Inst., Chinese Academy of Sciences;
(7) Bacillusthuringiensissusp.israelensis Bacillus thuringiensis subsp. israelensis ACCC---10038 ← Chengdu Hua Hong industrial corporation ← Taian Shandong biological study institute ← World Health Organization (WHO);
(8) Bacillusthuringienissubsp.thuringienisACCC-10035 ← China Agriculture Academe Fertilizer Institute; Original number: D25;
(9) Bacillusthuringiensissuhsp.berliner bacillus thuringiensis,Bt Bai Shi mutation ACCC-10022 ← China Agriculture Academe Fertilizer Institute; Original number?
(10) Bacillusthuringiensissubsp.Dendrolimus bacillus thuringiensis,Bt pine another name for Sichuan Province subspecies ACCC---10062 ← Soviet Union;
(11) Bacillusthuringienissubsp.sottoACCC-10033 ← Wuhan microbial pesticide factory ← Britain; Original number: 016;
According to (0.5-1.5) between the pure liquid of each Spawn incubation of selected every class bacterial classification: (0.5-1.5) ratio mixes.
4. antimicrobial composition insecticide according to claim 3, according to 1: 1 ratio mixing between the pure liquid of each Spawn incubation of selected every class bacterial classification.
5. the antimicrobial composition insecticide according to claim 3 or 4, described preparation is liquid or solid preparation.
6. the antimicrobial composition method for producing insecticide of one of claim 1-5, comprises the following steps:
(1) by various bacterial classification respectively cellar culture be the pure liquid of Spawn incubation:
(2) expansion of bacterium liquid is cultivated: carry out being mixed and made into every class mixed bacteria liquid according to described ratio by between the pure liquid of each Spawn incubation of every class fungi, actinomycetes and/or bacterium; Again described every class Mixed Microbes is mixed in described parts by weight of component ratio; Culture fluid is cultivated, and fermentation obtains finished product preparation.
7. antimicrobial composition method for producing insecticide according to claim 6, described step (2) be by mixed bacteria liquid and culture fluid in 1: the ratio of (8-10) is cultivated, described culture fluid furnish component weight ratio is: analysis for soybean powder 1-1.5, dextrin 0.3-0.9, peptone 0.2-0.4, dipotassium hydrogen phosphate 0.1-0.3, calcium carbonate 0.3-0.7, magnesium sulfate 0.2-0.6, ammonium sulfate 0.03-0.07, soya-bean oil 0.1-0.3; PH value is 7.2.
8. according to the antimicrobial composition method for producing insecticide of claim 6 or 7, described mixed bacteria liquid and culture fluid are cultivated in the ratio of 1: 9, described culture fluid furnish component weight ratio is: analysis for soybean powder 1.2, dextrin 0.6, peptone 0.3, dipotassium hydrogen phosphate 0.2, calcium carbonate 0.5, magnesium sulfate 0.4, ammonium sulfate 0.05, soya-bean oil 0.2.
9. antimicrobial composition method for producing insecticide according to claim 7, described mixed bacteria liquid adds in the liquid medium after purification; Put into seeding tank, at temperature 30 DEG C ± 1 DEG C, stirring per hour 200 times, cultivates 36-48 hour; Microscopy every gram bacteria containing amount reaches 2,000,000,000/gram time, stop cultivating, check and be registered seed bacterium liquid without miscellaneous bacteria.By qualified bacterium liquid in the ratio of 5%, add liquid medium in fermentation tank, cultivate 30-45 hour, on inspection without miscellaneous bacteria, bacterium liquid obviously becomes gemma and crystal, and content reaches 2,000,000,000/gram, namely can be finished product fermentation product.
10. according to the antimicrobial composition method for producing insecticide of claim 6 or 7, also comprise the preparation of step (3) solid fungicide: the calcium carbonate adding 8-10% in finished product bacterium liquid, stir 30 minutes; Plate-frame filtering; , stir 30 minutes, furnishing pulpous state; Spraying dry; Be pulverized and mixed evenly, obtain final product.
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