CN103749545B - Rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder - Google Patents
Rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder Download PDFInfo
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Abstract
The invention discloses a kind of rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder.Described wetting powder is grouped into by the one-tenth of following mass percentage: 8% ~ 20% rose dark brown Isaria and bacillus thuringiensis,Bt, 1 ~ 8% wetting agent, 1 ~ 12% dispersant, 0.1 ~ 0.5% uv-protector, and surplus is carrier.Described rose dark brown Isaria and the weight part ratio of bacillus thuringiensis,Bt are 10:0.4 ~ 10:1.Described rose dark brown Isaria is rose dark brown Isaria bacterial strain SCAU-IFCF02, and be preserved in China typical culture collection center on November 1st, 2013, deposit number is CCTCC NO:M2013526.Described bacillus thuringiensis,Bt is Kurstaki.This wetting powder fast, efficient, non agricultural chemical residuum, delay pest resistance to insecticide and produce, with environmental friendliness, especially have good control efficiency to Cruciferous Vegetable Pests such as diamond-back moths.
Description
Technical field
The present invention relates to mixed biologic pesticide field.More specifically, a kind of rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder is related to.
Background technology
The resistance problem of pest is one of global hot issues, is also the emphasis problem that agricultural science and technology worker pays close attention to always.At present to agricultural insect management mainly based on chemical control, the increase of Pesticide use amount and not scientifical use, cause pest resistance to insecticide day by day serious, the pest species producing resistance gets more and more, to natural enemy generation adverse effect.Meanwhile, the use agricultural chemicals of high strength, makes Practice for Pesticide Residue in Agricultural Products exceed standard, peasant's drug cost increases, contaminated environment and produce harmful effect etc. to the mankind and domestic animal, is unfavorable for the sustainable development of agricultural.Therefore, research and development high-efficiency environment friendly, agricultural chemicals that is pollution-free, noresidue have great importance to agricultural sustainable development.
Rose dark brown Isaria (
isaria fumosorosea) be a kind of important insect pathogenic fungus, be distributed widely in the torrid zone, subtropics and Temperate Region in China; Host range is wide, can comprise various vegetables, fruit tree and the Pests of Tea-Plants such as Homoptera, diptera, Lepidoptera, Semiptera, coleoptera and Hymenoptera by parasitism.Rose dark brown Isaria as a kind of living body biological agricultural chemicals, have be different from existing chemical insecticide the brand-new mechanism of action, to pest efficient, environmentally friendly, not easily the advantage such as to develop immunity to drugs, therefore become the focus of domestic and international biological control research.And, be one of important means in the sustainable production of non-polluted farm product with fungi microbe pesticide control plant pest, have broad application prospects.But fungi preparation is the prerequisite of its microbial pesticide application, since the sixties in 20th century, 10 kinds of rose dark brown Isaria formulation products are had to be found out and to obtain registration abroad, the commercial preparation come out in recent years had conidium preparation Biocon(Holland), AgoBiocontrol Bassiana 50(Colombia), Pae-sin(Mexico), Bemisin(Venezuela), blastopore preparation PFR-97 and blastopore and mycelial mix preparation PreFeRal.Although these existing rose dark brown Isaria preparations achieve good control efficiency to insect, the same with other biocontrol fungis, these preparation as insecticiding speed this shortcoming comparatively slow seriously constrains its application in field.
Bacillus thuringiensis,Bt (
bacillus thuringiensisbe called for short Bt) be use the biological insecticides the widest, output is maximum in the world, its main active is one or several insecticidal crystal proteins (insecticidal crystal proteins, ICPs), also known as delta-endotoxin, good specificity cytotoxicity is had to arthropodss such as the insects such as Lepidoptera, coleoptera, Hymenoptera, Homoptera and animals and plants nematode, acarians, and to non-target organism without adverse effect.Biotrol BTV is efficient, single-minded, to person poultry safety, that the advantage such as environmentally friendly makes Su Yun gold bar commercial preparation bacterium become consumption is in the world maximum, and the microorganism insecticide that the scope of application is wide, the most respond well, enjoys people to pay close attention to.But also there is quick-acting difference in commodity thuricade-1 in production control, the shortcomings such as, field lasting period insensitive to insect at advanced age short, easy generation resistance and recombinant strain genetic character instability, have become the restraining factors affecting bacillus thuringiensis,Bt further genralrlization and use.
As everyone knows, diamond-back moth (
plutella xylostellal.) be one of the important pests of worldwide brassicaceous vegetable, its host reaches more than 40 kinds, main harm brassicaceous vegetable, and year occurs many from generation to generation, and reproduction rate is high, and generation overlap phenomenon is serious.The current control to diamond-back moth is still based on chemical control, the continuous use of a large amount of chemical pesticide, under making diamond-back moth be in stronger medicament Selective Pressure all the time, add that again its reproduction speed is fast, cause it more and more higher to the resistance level of medicament, now become pesticide resistance the most serious and one of the insect of the most difficult control, have a strong impact on the seed output and quality of vegetables.At present, diamond-back moth creates resistance in various degree to more than 50 kinds of insecticides, and comprehensive regulation situation is very severe.
Summary of the invention
The technical problem to be solved in the present invention is the technical deficiency overcoming the Cruciferous Vegetable Pests controls such as existing diamond-back moth, provides one novel built insecticide fast and efficiently.
The object of the invention is to provide a kind of rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder.
Another object of the present invention is to provide the application of described wetting powder.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
The invention provides a kind of rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder, described wetting powder is grouped into by the one-tenth of following mass percentage: 8 ~ 20% rose dark brown Isaria and bacillus thuringiensis,Bts, 1 ~ 8% wetting agent, 1 ~ 12% dispersant, 0.1 ~ 0.5% uv-protector, surplus is carrier;
Described rose dark brown Isaria and the weight part ratio of bacillus thuringiensis,Bt are 10:0.4 ~ 10:1.More preferably, described rose dark brown Isaria and the weight part ratio of bacillus thuringiensis,Bt are 10:0.6 ~ 10:0.9.
Preferably, described wetting powder is grouped into by the one-tenth of following mass percentage: 10 ~ 18% rose dark brown Isaria and bacillus thuringiensis,Bts, 2 ~ 6% wetting agents, 2 ~ 10% dispersants, 0.2 ~ 0.4% uv-protector, and surplus is carrier; Described rose dark brown Isaria and the weight part ratio of bacillus thuringiensis,Bt are 10:0.6 ~ 10:0.9.
Described rose dark brown Isaria preferably adopt rose dark brown Isaria (
isaria fumosorosea(Wize) Brown & Smith) bacterial strain SCAU-IFCF02, this bacterial strain is preserved in China typical culture collection center on November 1st, 2013, deposit number is CCTCC NO:M 2013526, and preservation address is Wuhan University of Wuhan, China city.
The rose dark brown Isaria conidial powder of described rose dark brown Isaria to be spore content be 12,400,000,000/g.
Described bacillus thuringiensis,Bt is bacillus thuringiensis,Bt Kurstaki, and quality is 50000IU/mg, commercial.
Described wetting agent is octyl phenol APEO-10(OP-10), wetting agent Morwet EFW(alkylnaphthalene sulfonate and anion humectant mixture, Akzo Nobel N.V. produces), the mixture of one or more in fatty alcohol-polyoxyethylene ether (AEO-9).
Described dispersant is the mixture of one or more in fatty alcohol-polyoxyethylene ether, methylene two bitter edible plant sodium sulfonate, alkylnaphthalene sulfonate condensation polymer (Morwet D-425, Akzo Nobel N.V. produces).
Described uv-protector is the mixture of one or more in fluorescein sodium, xanthans, ascorbic acid.
Described carrier is one or both the mixture in talcum powder, diatomite.
Preferably, described wetting agent is one or both the mixture in octyl phenol APEO-10 or Morwet EFW.
Preferably described dispersant is one or both the mixture in methylene two bitter edible plant sodium sulfonate or Morwet D-425.
Preferably described uv-protector is ascorbic acid.
Preferably described carrier is talcum powder.
Present invention also offers described rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder and prepare the application in the preparation preventing and treating Homoptera, diptera, Lepidoptera, Semiptera, coleoptera or Hymenoptera insect.
Preferably, described insect is Cruciferous Vegetable Pests.
More preferably, described insect is diamond-back moth, red fire ant or Bemisia tabaci.
Preferably, described Cruciferous Vegetable Pests is diamond-back moth, cabbage caterpillar, aphid, Bemisia tabaci.
Preferably, the preparation method of rose dark brown Isaria conidial powder of the present invention is as follows:
S1. one-level solid culture, rose dark brown Isaria bacterial strain SCAU-IFCF02 is coated SDAY solid culture medium (SDAY medium is optimum medium), and cultivate in 24 DEG C, best incubation time is 15 days;
S2. secondary liquid shaken cultivation, use containing the spore on above-mentioned solid culture medium under the aseptic washing of 0.03% Tween-80, then is filtered with sterile gauze, makes 1 × 10
7the suspension of individual spore/mL, the above-mentioned inoculation of suspension liquid of 100uL in the Sa Shi culture fluid (being called for short SDB, is the suitableeest inoculation liquid) of 200mL, 24 DEG C, 170r/min shaken cultivation 72 hours;
S3. three grades are produced spore cultivation, low value old rice (common rice, be purchased from the Agricultural University Of South China market of farm produce) the quality distilled water such as to add, cool for subsequent use after 121 DEG C of moist heat sterilization 30min, the access of the bacterium liquid containing a large amount of mycelia obtained by S2 is on the rice of steam sterilizing, bacterium liquid is laid in after fully mixing with low value old rice and is lined with in the pallet of gauze, and covers with preservative film, 24 DEG C of dark culturing 6 days.When media surface covers with mycelia, throw off preservative film continue cultivation 4 days, make the large volume production spore of mycelia, results conidial powder, vacuum cooling drying 24 hours, dried Conidial storage is in sterile chamber, and 4 DEG C save backup.The conidial powder spore content obtained is 12,400,000,000/g, and spore rate of living is 97.8%, and water content is 2.56%, and outward appearance is light rose dark brown powder.
Wherein, described in S1, SDAY solid culture medium compound method is as follows:
Glucose 40g/L, peptone 10g/L, yeast extract 20g/L, agar 15 ~ 20g/L, H
20 1L, stirring and evenly mixing, then packing, sterilizing, for subsequent use.
Described in S2, Sa Shi culture fluid compound method is as follows:
Glucose 40g/L, peptone 10g/L, yeast extract 20g/L, H
201L, stirring and evenly mixing, then packing, sterilizing, for subsequent use.
Inventor herein is separated the rose dark brown Isaria bacterial strain SCAU-IFCF02 obtained, and has low-down lethal concentration and the fastest lethal speed to diamond-back moth, overcomes common fungi microbe agricultural insecticide speed these restraining factors slow.Therefore, the present invention infects body conidium for active component with the common of rose dark brown Isaria bacterial strain SCAU-IFCF02, be prepared into rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder, can lay the foundation for the industrialization of rose dark brown Isaria bacterial strain SCAU-IFCF02, for the Field information of this bacterial strain provides theoretical direction, have a good application prospect.
The collocation of rose dark brown Isaria and bacillus thuringiensis,Bt uses, and is used for preventing and treating the Cruciferous Vegetable Pests such as diamond-back moth, there is no report and application at present.The collocation of different pesticide species and different insect killing substance uses, and effectively can improve control efficiency, reduces dosage, reduces costs, and delaying the generation of pest resistance to insecticide, extend the service life of medicament, is the important means of integrated pest management.The present inventor is on the basis of a large amount of indoor bioassay and field medicine effect test, find that bacillus thuringiensis,Bt and rose dark brown Isaria SCAU-IFCF02 bacterial strain have good compatibility, and rose dark brown Isaria provided by the invention and bacillus thuringiensis,Bt wetting powder are as a kind of biological insecticides, alone rose dark brown Isaria and bacillus thuringiensis,Bt are better than to the desinsection speed of target pest and successful, there is different desinsection modes and desinsection object, the two carries out checking the novel compounded pesticides of acquisition, expand insecticidal spectrum, improve insecticide efficiency, slow down pest resistance to insecticide to produce, reduce the use of chemical pesticide.Prepared insecticide has the brand-new mechanism of action being different from existing insecticide, and insecticidal effect is remarkable, and the toxic action especially for Cruciferous Vegetable Pests diamond-back moth is remarkable, has environmental friendliness, to the feature of person poultry safety simultaneously.Therefore, the present invention infects body conidium and bacillus thuringiensis,Bt for active component with rose dark brown Isaria bacterial strain SCAU-IFCF02 common, be prepared into rose dark brown cluster bacillus thuringiensis,Bt spore wetting powder, achieve extraordinary pest control effect, for the industrialization of rose dark brown Isaria preparation lays the foundation, for the Field information of said preparation provides theoretical direction.
The present invention has following beneficial effect:
In rose dark brown Isaria provided by the invention and bacillus thuringiensis,Bt wetting powder, bacillus thuringiensis,Bt and rose dark brown Isaria spore have good compatibility, synergistic effect is obvious, alone rose dark brown Isaria and bacillus thuringiensis,Bt are better than to the desinsection speed of target pest and successful, reach that a medicine is controlled, the effect of quick-acting guarantor's liquid more.Desinsection collection of illustrative plates expands, and except control diamond-back moth, can also prevent and treat other Cruciferous Vegetable Pests such as cabbage caterpillar, aphid, Bemisia tabaci, and this wetting powder can superinfection, successful on the non-environmental pollution control and lasting integrated control of insect.
In addition, wettable powder agent producing process of the present invention is simple, significantly can reduce the usage amount of agricultural chemicals, delay pest resistance to insecticide, reduce residue of pesticide, to person poultry safety, environmentally friendly, to agricultural production and income, improve the ecological environment, developing environmental protection agricultural, having great importance to realizing the strategy of sustainable development.
Embodiment
Further illustrate the present invention below in conjunction with specific embodiment, but embodiment does not limit in any form to the present invention.Unless stated otherwise, the present invention adopts reagent, method and apparatus are the art conventional reagent, method and apparatus.
Unless stated otherwise, following examples agents useful for same and raw material are conventional method preparation or commercial.
The rose dark brown Isaria conidial powder of embodiment of the present invention rose dark brown used Isaria to be spore content be 12,400,000,000/g.
The assay method of the present invention to spore germination rate is as follows: spore suspension is inoculated into fungi germination fluid (glucose 20g/L, peptone 5g/L, H
20 1L) in, 24 DEG C, 170 r/min, microscopy statistics spore rate alive after 18h.Wetting time measures according to GB/T5451-2001 standard method.Suspensibility is pressed GB/T 14825-2006 standard method and is measured.PH value is pressed GB/T1600-93 standard method and is measured.Fineness is measured by laser particle distribution instrument.Data statistic analysis: all experimental data DPS data handling systems are analyzed, computation of mean values and standard error, the average significance of difference is carried out multiple analysis by Duncan duncan's new multiple range method and is compared.
Sterile water containing 0.03% Tween-80 used in embodiment, described 0.03% refers to volume ratio.
the preparation of embodiment 1 rose dark brown Isaria conidial powder
The present embodiment adopts solid-liquid double-phase fermented and cultured, and prepare rose dark brown Isaria conidial powder, preparation process is as follows:
S1. one-level solid culture, get 50uL rose dark brown Isaria bacterial strain SCAU-IFCF02 original bacteria liquid and coat (SDAY medium is optimum medium) on SDAY solid culture medium, cultivate in 24 DEG C, best incubation time is 15 days;
S2. secondary liquid shaken cultivation, use containing the spore on SDAY solid culture medium under the aseptic washing of 0.03% Tween-80, then is filtered with double gauze, makes 1 × 10
7the suspension of individual spore/mL, the above-mentioned inoculation of suspension liquid of 100uL in the Sa Shi culture fluid (being called for short SDB, is the suitableeest inoculation liquid) of 200mL, 24 DEG C, 170r/min shaken cultivation 72 hours;
S3. three grades are produced spore cultivation, low value old rice (common rice, be purchased from the Agricultural University Of South China market of farm produce) the quality distilled water such as to add, cool for subsequent use after 121 DEG C of moist heat sterilization 30min, on the rice of the access of the bacterium liquid containing a large amount of mycelia sterilizing that S2 is obtained, bacterium liquid is laid in after fully mixing with low value old rice and is lined with in the pallet of gauze, and covers with preservative film, 24 DEG C of dark culturing 6 days.When media surface covers with mycelia, throw off preservative film continue cultivation 4 days, make the large volume production spore of mycelia, results conidial powder, vacuum cooling drying 24 hours, dried Conidial storage is in sterile chamber, and 4 DEG C save backup.
The conidial powder spore content obtained is 12,400,000,000/g, and spore rate of living is 97.8%, and water content is 2.56%, and outward appearance is light rose dark brown powder.
Described in step S1, SDAY solid culture medium compound method is as follows:
Glucose 40g/L, peptone 10g/L, yeast extract 20g/L, agar 15 ~ 20g/L, H
20 1L, stirring and evenly mixing, then packing, sterilizing, for subsequent use.
Described in step S2, Sa Shi culture fluid compound method is as follows:
Glucose 40g/L, peptone 10g/L, yeast extract 20g/L, H
2o 1L, stirring and evenly mixing, then packing, sterilizing, for subsequent use.
the preparation of embodiment 2 rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder
1, preparation method:
S1. rose dark brown Isaria conidial powder and the Bacillus thuringiensis (commercial) of embodiment 1 preparation is accurately taken;
S2. various auxiliary agent is joined in low-temperature airflow cracker, control the temperature of air-flow crushing below 10 DEG C, for subsequent use after pulverizing; Preferably according to 325 order finess disintegratings,
S3. proportionally S2 being pulverized the various agent mixture obtained joins in carrier, stirs, then adds S1 load weighted rose dark brown Isaria conidial powder and Bacillus thuringiensis, namely obtain the wetting powder of mixture after mixing.
2, the wetting powder prepared by the present embodiment comprises the constituent of following percentage by weight: rose dark brown Isaria 9.2%, bacillus thuringiensis,Bt 0.8%, OP-10 3%, Morwet EFW 2%, methylene two bitter edible plant sodium sulfonate 3%, Morwet D-425 5%, ascorbic acid 0.3%, talcum powder 76.7%.
The performance indications of the wetting powder 3, prepared by the present embodiment measure
Testing result shows, product performance index is excellent, rose dark brown Isaria spore germination rate >=85; Bacillus thuringiensis Spore germination rate is 84.17%, wetability <1min; Quality suspensibility is 72.3; The suspensibility of spore is 85.5, and moisture is 2.56; Fineness (crossing 325 mesh standard sieves) >=98; PH value is 6.2.The performance indications of the wetting powder prepared meet the requirement of pesticidal preparations commodity.
the preparation of embodiment 3 rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder
1, preparation method is with embodiment 2.
2, the wetting powder prepared by the present embodiment comprises the constituent of following percentage by weight: rose dark brown Isaria 11.3%, bacillus thuringiensis,Bt 0.7%, AEO-9 4%, Morwet EFW 2%, fatty alcohol-polyoxyethylene ether 2%, Morwet D-425 7%, fluorescein sodium 0.3%, diatomite 45.9%, talcum powder 26.8%.
The performance indications of the wetting powder 3, prepared by the present embodiment measure
Testing result shows, product performance index is excellent, rose dark brown Isaria spore germination rate >=85; Bt gemma germination rate is 82.17%, wetability <1min; Quality suspensibility is 71.9; The suspensibility of spore is 86.7, and moisture is 2.96; Fineness (crossing 325 mesh standard sieves) >=98; PH value is 6.5.The performance indications of the wetting powder prepared meet the requirement of pesticidal preparations commodity.
the preparation of embodiment 4 rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder
1, preparation method is with embodiment 2.
2, the wetting powder prepared by the present embodiment comprises the constituent of following percentage by weight: rose dark brown Isaria 13.8%, bacillus thuringiensis,Bt 1.2%, OP-10 2%, Morwet EFW 2%, fatty alcohol-polyoxyethylene ether 2%, methylene two bitter edible plant sodium sulfonate 3%, Morwet D-425 6%, xanthans 0.5%, talcum powder 69.5%.
The performance indications of the wetting powder 3, prepared by the present embodiment measure
Testing result shows, product performance index is excellent, rose dark brown Isaria spore germination rate >=85; Bt gemma germination rate is 86.17%, wetability <1min; Quality suspensibility is 74.3; The suspensibility of spore is 87.5, and moisture is 3.06; Fineness (crossing 325 mesh standard sieves) >=98; PH value is 6.4.The performance indications of the wetting powder prepared meet the requirement of pesticidal preparations commodity.
the preparation of embodiment 5 rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder
1, preparation method is with embodiment 2.
2, the wetting powder prepared by the present embodiment comprises the constituent of following percentage by weight: rose dark brown Isaria 17%, bacillus thuringiensis,Bt 1%, OP-10 5%, Morwet EFW 3%, fatty alcohol-polyoxyethylene ether 2%, methylene two bitter edible plant sodium sulfonate 4%, Morwet D-425 5%, ascorbic acid 0.5%, talcum powder 30%, diatomite 32.5%.
The performance indications of the wetting powder 3, prepared by the present embodiment measure
Testing result shows, product performance index is excellent, rose dark brown Isaria spore germination rate >=85; Bt gemma germination rate is 84.17%, wetability <1min; Quality suspensibility is 72.3; The suspensibility of spore is 85.5, and moisture is 2.56; Fineness (crossing 325 mesh standard sieves) >=98; PH value is 6.2.The performance indications of the wetting powder prepared meet the requirement of pesticidal preparations commodity.
the preparation of embodiment 6 rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder
1, preparation method is with embodiment 2.
2, the wetting powder prepared by the present embodiment comprises the constituent of following percentage by weight: rose dark brown Isaria 18%, bacillus thuringiensis,Bt 2%, OP-10 4%, Morwet EFW 4%, fatty alcohol-polyoxyethylene ether 2%, methylene two bitter edible plant sodium sulfonate 5%, Morwet D-425 5%, ascorbic acid 0.4%, talcum powder 59.6%.
The performance indications of the wetting powder 3, prepared by the present embodiment measure
Testing result shows, product performance index is excellent, rose dark brown Isaria spore germination rate >=85; Bt gemma germination rate is 85.37%, wetability <1min; Quality suspensibility is 74.3; The suspensibility of spore is 85.5, and moisture is 2.78; Fineness (crossing 325 mesh standard sieves) >=98; PH value is 6.5.The performance indications of the wetting powder prepared meet the requirement of pesticidal preparations commodity.
the evaluating drug effect of embodiment 7 rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder
1, for examination material
(1) for examination insect: diamond-back moth
plutella xylostella(L.) 2 instar larvaes.
(2) reagent agent: rose dark brown Isaria prepared by embodiment 2 and bacillus thuringiensis,Bt wetting powder.
Inventor tests through Toxicity Determination, result shows, rose dark brown Isaria prepared by above-described embodiment 2 ~ 6 and bacillus thuringiensis,Bt wetting powder all have extraordinary control efficiency to diamond-back moth, when wettable powder agent concentration is dilution 500, from connect the 1st day diamond-back moth after bacterium just death, the corrected mortality of the 4th day diamond-back moth is all between 90% ~ 100%.
Because data are too many, the data of the wetting powder control diamond-back moth that the present invention only provides embodiment 2 to prepare, are enough to illustrate that rose dark brown Isaria provided by the invention and bacillus thuringiensis,Bt wetting powder have good control efficiency to insect.
(3) medicament is contrasted: the rose dark brown Isaria wetting powder single dose prepared with embodiment 2 preparation method and bacillus thuringiensis,Bt wetting powder single dose.
(4) for studying thing: cabbage mustard (
b. alboglabrabailey).
2, rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder are to the Toxicity Determination of diamond-back moth
(1) experimental technique
Rose dark brown Isaria and bacillus thuringiensis,Bt wettable powder dilution agent 500 times, 1000 times, 2000 times and 5000 times of four concentration.
Dip method is adopted to carry out biologicall test: to choose Individual Size, diamond-back moth 2 instar larvae that vigor is consistent, to choose on clean cabbage mustard blade for examination larva with banister bruss, then blade and larva are put into the wetting powder of different extension rate, choose after dipping 10s, after filter paper larva being placed in sterilizing blots excessive moisture, move on blade, put into the culture dish (diameter is 9cm) that bottom is lined with moistening filter paper, culture dish preservative film seals moisturizing, film is pricked hole ventilation, be placed in artificial intelligence case, 24 DEG C, relative moisture (RH) 90%, periodicity of illumination is illumination: dark=14:10.Often process 20 larvas, repeat 5 times, totally 100 larvas.With wetting powder basic components (not comprising the wetting powder of rose dark brown Isaria conidial powder and bacillus thuringiensis,Bt) for blank, dilute 500 times for medicament contrast with rose dark brown Isaria wetting powder single dose and bacillus thuringiensis,Bt wetting powder single dose.Start after 24 hours to observe, changed fresh cabbage leaves depending on ware intra vane by the situation of taking food, record susceptible symptom and dead borer population every day, Continuous Observation 6 days, calculate lethality and corrected mortality.
Lethality (%)=dead borer population/each concentration process examination worm sum × 100
Corrected mortality (%)=(process lethality-contrast lethality)/(1-contrasts lethality) × 100
(2) Toxicity Determination result
Rose dark brown Isaria and the Toxicity Determination result of bacillus thuringiensis,Bt wetting powder to diamond-back moth as shown in table 1.
Table 1 rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder are to diamond-back moth 2 instar larvae toxicity test result
As shown in table 1, namely after medication 24h there is obvious death in diamond-back moth, after 24h, 48h, 72h, 96h and 120h, rose dark brown Isaria of the present invention and bacillus thuringiensis,Bt wetting powder are all significantly higher than single dose and blank to diamond-back moth 2 instar larvae lethality, and corrected mortality is up to 100%, indicate rose dark brown Isaria of the present invention and bacillus thuringiensis,Bt wetting powder has extraordinary control efficiency to diamond-back moth.
3, the field trial of diamond-back moth prevented and treated by rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder
(1) test method
Experiment arranges 15 communities, each plot area 16 square metres altogether, and random district group arranges.Rose dark brown Isaria and bacillus thuringiensis,Bt wettable powder dilution agent 500 times, 1000 times, 2000 times three concentration, 500 times are diluted for medicament contrast with bacillus thuringiensis,Bt wetting powder and rose dark brown Isaria wetting powder single dose, be treated to blank with clear water, each process repeats 3 times.Community even spraying is divided with commercially available knapsack hand sprayer.Per hectare amount of liquid medicine 900kg, every community water 1.45kg.
Test employing 5 methods are investigated, the sampling of 5, every community, often 2 strain fixed point investigation, investigation insect population radix before medicine, 1,3,7 day residual borer population of investigation diamondback moth larvae after medicine, calculate Revision insect recluced rate and relative control effect, the analysis of all data all adopts DPS software to complete.
Borer population × 100 alive before Revision insect recluced rate (%)=(borer population of living after borer population-medicine of living before medicine)/medicine
Control efficiency (%)=(PT-CK)/(100-CK) × 100
In formula: CK is blank district Revision insect recluced rate, PT is chemicals treatment district Revision insect recluced rate.
(2) result of the test
Rose dark brown Isaria and the field efficacy of bacillus thuringiensis,Bt wetting powder to diamond-back moth as shown in table 2.
Table 2 rose dark brown Isaria bacillus thuringiensis,Bt wetting powder control diamond-back moth field control effectiveness test result
As shown in table 2, rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder have significant control efficiency to diamond-back moth, and when using dosage is dilution 500 times, after medicine, the preventive effect of the 1st day, 3 days and 7 days is respectively 52.47%, 69.72%, 89.93%; When its using dosage is dilution 1000 times, after medicine, the preventive effect of the 1st day, 3 days and 7 days is respectively 52.86%, 64.94%, 85.40%, all good than contrast effect of chemical control.As the environmentally safe microbial pesticide of one, rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder have great exploitation prospect, have to Cruciferous Vegetable Pests such as diamond-back moths the persistence that extraordinary preventive effect becomes reconciled.
In addition, according to duration of test field observation, each treatment region is normal for examination plant growth, does not find any harmful effect, also has no adverse effects to other non-target organisms.And found by data survey, each treatment region has cabbage caterpillar, Bemisia tabaci mixing harm, after dispenser, the quantity of cabbage caterpillar and Bemisia tabaci also significantly declines, illustrate that rose dark brown Isaria of the present invention and bacillus thuringiensis,Bt wetting powder have good control efficiency to other insects such as cabbage caterpillar and Bemisia tabaci simultaneously, desinsection collection of illustrative plates is larger, can control by a medicine, and this wetting powder can superinfection, successful on the non-environmental pollution control and lasting integrated control of insect more.
Claims (4)
1. a rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder, it is characterized in that, described wetting powder is grouped into by the one-tenth of following mass percentage: 8 ~ 20% rose dark brown Isaria and bacillus thuringiensis,Bts, 1 ~ 8% wetting agent, 1 ~ 12% dispersant, 0.1 ~ 0.5% uv-protector, surplus is carrier;
Described rose dark brown Isaria and the weight ratio of bacillus thuringiensis,Bt are 10:0.4 ~ 10:1;
Described rose dark brown Isaria is rose dark brown Isaria bacterial strain SCAU-IFCF02, and this bacterial strain is preserved in China typical culture collection center on November 1st, 2013, and deposit number is CCTCC NO:M 2013526;
Described bacillus thuringiensis,Bt is bacillus thuringiensis,Bt Kurstaki;
Described wetting agent is octyl phenol APEO-10, and described dispersant is fatty alcohol-polyoxyethylene ether, and described uv-protector is ascorbic acid, and described carrier is talcum powder.
2. rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder according to claim 1, it is characterized in that, described wetting powder is grouped into by the one-tenth of following mass percentage: 10 ~ 18% rose dark brown Isaria and bacillus thuringiensis,Bts, 2 ~ 6% wetting agents, 2 ~ 10% dispersants, 0.2 ~ 0.4% uv-protector, surplus is carrier;
Described rose dark brown Isaria and the weight ratio of bacillus thuringiensis,Bt are 10:0.6 ~ 10:0.9.
3. rose dark brown Isaria and bacillus thuringiensis,Bt wetting powder according to claim 1 or 2, is characterized in that, the rose dark brown Isaria conidial powder of described rose dark brown Isaria to be spore content be 12,400,000,000/g.
4. the application in the preparation preventing and treating diamond-back moth prepared by the arbitrary described rose dark brown Isaria of claims 1 to 3 and bacillus thuringiensis,Bt wetting powder.
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| CN104642392B (en) * | 2015-01-13 | 2018-05-01 | 华南农业大学 | A kind of fumosorosea bacterium conidium wettable powder and its application |
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