CN103960288A - Application of mixed formula preparation to control on nilaparvata lugens stal - Google Patents
Application of mixed formula preparation to control on nilaparvata lugens stal Download PDFInfo
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- CN103960288A CN103960288A CN201410128645.3A CN201410128645A CN103960288A CN 103960288 A CN103960288 A CN 103960288A CN 201410128645 A CN201410128645 A CN 201410128645A CN 103960288 A CN103960288 A CN 103960288A
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- streptomyces diastatochromogenes
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- imidacloprid
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Abstract
The invention relates to the technical field of microbiology, particularly to an application of a mixed formula preparation to control on nilaparvata lugens stal. Streptomyces diastatochromogenes is named streptomyces diastatochromogenes D. The strain is preserved in the CGMCC (China General Microbiological Culture Collection Center) of CCCCM(China Committee for Culture Collection of Microorganisms)on May 15th, 2007, and the preservation registration number is CGMCC No.2060. Metabolic products having an inhibition effect on nilaparvata lugens stal yeast-likesymbiotic bacteria can be obtained throughfermentation and extraction of the streptomyces diastatochromogenes D.The application is most significantly characterized in that the metabolic products of the streptomyces diastatochromogenes D and imidacloprid are mixed in certainproportion to prepare a new formula, the usage amount of a pesticide can be obviously reduced when the mixed formula is used for controlling the nilaparvata lugens stal, the control effect on the nilaparvata lugens stal can be effectively improved, and the development of the drug resistance of the nilaparvata lugens stal can be delayed.
Description
Technical field
The present invention relates to microbial technology field, particularly relate to a kind of streptomyces diastatochromogenes D metabolite and Imidacloprid mixing formula preparation.
Background technology
Brown planthopper
one of important pests of paddy rice, the mode of mainly inhaling phloem sap with thorn and the propagating Rice Virus rice plant of causing harm.During the large generation of insect pest, there will be the withered lodging of large-area rice strain, cause grain drop in production, even total crop failure.In production, chemical pesticide is the Main Means of control brown planthopper.But due to a large amount of frequent use of chemical insecticide, brown planthopper has produced pesticide resistance to many chemical insecticides, causes control efficiency obviously to decline.Therefore, research and application reduce that Brown Planthopper occurs and the new tool that causes harm is extremely urgent.
The symbiosis of insect and microorganism is a kind of ubiquitous phenomenon, and both have formed inseparable relation during evolution.Insect provides stable microhabitat and nutrition for fungal component, and insect relies on some the important nutrient component lacking in fungal component purified diet, fungal component also has certain effect to the metabolic detoxification of insect simultaneously, as being the synthetic important nutrient component of insect, participate in nitrogen cycle, the interior allogenic material of degraded insect bodies etc.Disappearance fungal component can directly cause that insect growth breeding is obstructed, lethality rising etc.Research shows, in brown planthopper body, in stomach fat body, also has multiple types yeast fungal component, Parasaccharomyces Ascomycotina (Ascomycotina), and gang pyrenomycetes (Pyrenomycetes), Candida belongs to.Brown planthopper class yeast fungal component can be that host is synthetic provides the multiple nutritional components such as essential amino acid, steroid material and protein, made up the imbalance that brown planthopper takes food trophic structure, guaranteed normal growth and the breeding of brown planthopper; Yeast-like endosymbiote disappearance can cause brown planthopper growth retardation, arrest of development, even dead.Since brown planthopper and class yeast fungal component relation in growth and breeding are so tight, by the means of " indirectly preventing and treating brown planthopper thereby suppress class yeast fungal component ", controlling that brown planthopper occurs and reduce that it causes harm will be a kind of important means of preventing and treating from now on brown planthopper.Because suppress the object of controlling brown planthopper that reaches that fungal component in brown planthopper body not only can be direct or indirect, but also can effectively improve current insecticide control efficiency, delay the drug-fast development of brown planthopper.
The present invention is mixed with into new formula with common insecticides Imidacloprid according to certain ratio by streptomyces diastatochromogenes D metabolite, and has studied inhibitory action and the insecticidal effect to brown planthopper of this formula to class yeast fungal component in brown planthopper body.The control that this invention is Brown Planthopper provides new more efficiently method.
Summary of the invention
The object of the invention is for brown planthopper, conventional chemical insecticide to be developed immunity to drugs, the deficiency that causes chemical control successful to decline, provide a kind of streptomyces diastatochromogenes D metabolite that utilizes to suppress class yeast fungal component growth in brown planthopper body, and then cause the weak even dead new approaches of brown planthopper; Another object of the present invention has been to provide a kind of new method and new medicament of preventing and treating brown planthopper.
Streptomyces diastatochromogenes of the present invention (Streptomyces diastatochromogenes), names as streptomyces diastatochromogenes D.The center preservation of this bacterial strain Yi China Committee for Culture Collection of Microorganisms's common micro-organisms, No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, preservation address, preservation date on May 25th, 2007, preservation registration number CGMCC No.2060.Streptomyces diastatochromogenes D of the present invention, extracts and can obtain the inhibited metabolite of brown planthopper class yeast fungal component through fermentation.
The object of the invention is achieved through the following technical solutions:
A mixing formula preparation of preventing and treating brown planthopper, carries out according to the following steps:
(1) activation culture of bacterial classification: the streptomyces diastatochromogenes D bacterial classification that separation is preserved moves in vitro on Gause I inclined-plane, under 28 ℃ ± 1 ℃ condition, is cultured to test tube slant and covers with spore;
(2) fermented and cultured: fermentation culture is to contain soybean cake powder 20g in every 1000mL, corn flour 15g, wheat bran 10g, NH
4nO
33g, (NH
4)
2sO
45g, CaCO
35g, MgSO
41g, sterilizing, to be cooled to 40 ℃, under aseptic condition, be seeded in fermentation culture, under 28 ℃ ± 1 ℃ condition, cultivate 5 days;
(3) fermentation product extracts: ferment complete, centrifugal collection fermented supernatant fluid, learns from else's experience after Vacuum Concentration and obtain medicinal extract with conventional organic solvent extracting;
(4) liquid preparation: medicinal extract and common insecticides proportioning that step (3) is obtained, for the control of brown planthopper.
Beneficial effect of the present invention: the invention provides a kind of new method of utilizing streptomyces diastatochromogenes D metabolite and Imidacloprid mixing formula control brown planthopper.
Embodiment
The preparation method of embodiment 1:(streptomyces diastatochromogenes D metabolite)
Carry out according to the following steps:
(1) activation culture of bacterial classification: the streptomyces diastatochromogenes D bacterial classification that separation is preserved moves in vitro on Gause I inclined-plane, under 28 ℃ ± 1 ℃ condition, is cultured to test tube slant and covers with spore;
(2) fermented and cultured: fermentation culture is to contain soybean cake powder 20g in every 1000mL, corn flour 15g, wheat bran 10g, NH
4nO
33g, (NH
4)
2sO
45g, CaCO
35g, MgSO
41g, sterilizing, to be cooled to 40 ℃, under aseptic condition, inoculate a metal platinum filament ring streptomyces diastatochromogenes D spore to fermentation culture, under 28 ℃ ± 1 ℃ condition, cultivate 5 days;
(3) fermentation product extracts: centrifugal collection zymotic fluid after fermentation, add the extraction of equal-volume ethyl acetate, and take off layer, use again equal-volume extracting n-butyl alcohol, get upper strata, under vacuum condition, be concentrated into medicinal extract, this medicinal extract is streptomyces diastatochromogenes D metabolite, for control brown planthopper.
Streptomyces diastatochromogenes D metabolite described in following examples 2 is embodiment 1 products made thereby.
The preparation of embodiment 2:(streptomyces diastatochromogenes D metabolite and Imidacloprid mixing formula)
Imidacloprid water dispersible granule (active ingredient mass percent is 70%) is produced by Beyer Co., Ltd, imidacloprid water dispersible granule and streptomyces diastatochromogenes D metabolite are dissolved with running water respectively, mix, being made into final concentration is 0.2g/L streptomyces diastatochromogenes D metabolite and 0.01g/L Imidacloprid mixing formula.
The application in control brown planthopper nymph of embodiment 3:(streptomyces diastatochromogenes D metabolite and Imidacloprid mixing formula)
Carry out according to the following steps:
(1) the shooting stage TN1 paddy rice that tillers, removes part root and base portion leaf sheath afterwash, naturally dries;
(2) with clear water, compare, spraying final concentration is 0.2g/L streptomyces diastatochromogenes D metabolite and 0.01g/L Imidacloprid confection, requires medicament droplet to be evenly distributed, coverage rate is high, pharmaceutical quantities is suitable, does not produce to run off be advisable with moistening object surface, naturally dries;
(3) rice seedling after step (2) is processed is put into the Boiling tube of prior injection 20ml paddy rice culture fluid, 3 rice seedlings of every pipe, access 3 age 30 of brown planthopper nymphs, the rice seedlings spraying after clear water of usining is processed as negative control, spray 0.04g/L Imidacloprid liquid and be treated to positive control, each is processed 3 times and repeats;
(4) in processing, within latter 1 day, 2 days and 4 days, measure brown planthopper parachorium bacterium quantity respectively, and add up brown planthopper lethality.Wherein brown planthopper parachorium bacterium quantitative measurement method is: get respectively each process in 3 of the brown planthoppers of survival, alcohol disinfecting with 75%, rinsed with sterile water, remove wing, head and chest, belly is put into 1.5ml centrifuge tube, add 0.9% physiological saline 50 μ l, homogenate, then get 2 μ l homogenate drops on blood cell counting plate, the fragmentation that closes the lid, in the lower statistics of microscope (16 * 40) fungal component quantity.
The application in the female adult of control brown planthopper of embodiment 4:(streptomyces diastatochromogenes D metabolite and Imidacloprid mixing formula)
Carry out according to the following steps:
(1) the shooting stage TN1 paddy rice that tillers, removes part root and base portion leaf sheath afterwash, naturally dries;
(2) with clear water, compare, spraying final concentration is 0.2g/L streptomyces diastatochromogenes D metabolite and 0.01g/L Imidacloprid confection, requires medicament droplet to be evenly distributed, coverage rate is high, pharmaceutical quantities is suitable, does not produce to run off be advisable with moistening object surface, naturally dries;
(3) rice seedling after step (2) is processed is put into the Boiling tube of prior injection 20ml culture fluid, 4 rice seedlings of every pipe, 30 of the female adults of brachypterism that access has just been sprouted wings, the rice seedlings of usining after spraying is clearly processed as negative control, spray 0.04g/L Imidacloprid liquid and be treated to positive control, each is processed 3 times and repeats;
(4) in processing, within latter 1 day, 2 days and 4 days, measure the female worm brown planthopper of brachypterism parachorium bacterium quantity respectively, and add up brown planthopper lethality.Wherein brown planthopper parachorium bacterium quantitative measurement method is: get respectively each process in 3 of the female worms of brachypterism of survival, alcohol disinfecting with 75%, rinsed with sterile water, remove wing, head and chest, belly is put into 1.5ml centrifuge tube, add 0.9% physiological saline 50 μ l, homogenate, then get 2 μ l homogenate drops on blood cell counting plate, the fragmentation that closes the lid, in the lower statistics of microscope (16 * 40) fungal component quantity.
Test example: (streptomyces diastatochromogenes D metabolite of the present invention and Imidacloprid mixing formula and conventional chemical pesticide control Piglet s colibacillosis)
Result of the test is in Table 1 and table 2.As seen from Table 1 and Table 2, utilize streptomyces diastatochromogenes D metabolite and Imidacloprid mixing formula obviously to suppress the quantity of brown planthopper parachorium bacterium, thereby cause that brown planthopper lethality rises.
Table 1 streptomyces diastatochromogenes D metabolite and the control efficiency of Imidacloprid confection to brown planthopper nymph
Table 2 streptomyces diastatochromogenes D metabolite and the control efficiency of Imidacloprid confection to the female adult of brown planthopper
Streptomyces diastatochromogenes D metabolite and Imidacloprid decrement formula are better than conventional chemical liquid or similar to the control efficiency of brown planthopper nymph and female adult.Use streptomyces diastatochromogenes D metabolite of the present invention and Imidacloprid mixing formula control brown planthopper can obviously reduce utilization of pesticides amount, effectively improve the control efficiency of brown planthopper, delay the drug-fast development of brown planthopper.
Claims (5)
1. the application of mixing formula preparation in control brown planthopper, this mixing formula preparation comprises streptomyces diastatochromogenes D metabolite and Imidacloprid.
2. mixing formula preparation claimed in claim 1, wherein said streptomyces diastatochromogenes D metabolite is made by strain activation and culture, fermented and cultured, fermentation product extracting method.
3. mixing formula preparation claimed in claim 1, the percentage by weight of wherein said streptomyces diastatochromogenes D metabolite and Imidacloprid is 10: 1-20: 1.
4. mixing formula preparation claimed in claim 1, is obtained by following methods:
(1) activation culture of bacterial classification: the streptomyces diastatochromogenes D bacterial classification that separation is preserved moves to and is in vitro cultured to test tube slant and covers with spore, described streptomyces diastatochromogenes D Yi China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, preservation registration number is CGMCC No.2060;
(2) fermented and cultured: fermentation medium is soybean cake powder, corn flour, wheat bran, NH
4nO
33g, (NH
4)
2sO
4, CaCO
3, MgSO
4, sterilizing, treats coldly, inoculates streptomyces diastatochromogenes D spore under aseptic condition to fermentation medium, cultivates;
(3) fermentation product extracts: ferment complete, and centrifugal collection fermented supernatant fluid, extraction, takes off layer, then extracts, and gets upper strata, under vacuum condition, is concentrated into medicinal extract, and this medicinal extract is streptomyces diastatochromogenes D metabolite;
(4) mixing formula preparation: imidacloprid water dispersible granule and streptomyces diastatochromogenes D metabolite are dissolved with running water respectively, mix.
5. mixing formula preparation claimed in claim 4, is obtained by following methods:
(1) activation culture of bacterial classification: the streptomyces diastatochromogenes D bacterial classification that separation is preserved moves in vitro on Gause I inclined-plane, under 28 ℃ ± 1 ℃ condition, be cultured to test tube slant and cover with spore, described streptomyces diastatochromogenes D Yi China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, preservation registration number is CGMCC No.2060;
(2) fermented and cultured: fermentation culture is to contain soybean cake powder 20g in every 1000mL, corn flour 15g, wheat bran 10g, NH
4nO
33g, (NH
4)
2sO
45g, CaCO
35g, MgSO
4lg, sterilizing, to be cooled to 40 ℃, under aseptic condition, inoculate a metal platinum filament ring streptomyces diastatochromogenes D spore to fermentation culture, under 28 ℃ ± 1 ℃ condition, cultivate 5 days;
(3) fermentation product extracts: ferment complete, centrifugal collection fermented supernatant fluid, adds the extraction of equal-volume ethyl acetate, take off layer, then use equal-volume extracting n-butyl alcohol, get upper strata, under vacuum condition, be concentrated into medicinal extract, this medicinal extract is streptomyces diastatochromogenes D metabolite;
(4) mixing formula preparation: imidacloprid water dispersible granule (active ingredient mass percent is 70%) is produced by Beyer Co., Ltd, imidacloprid water dispersible granule and streptomyces diastatochromogenes D metabolite are dissolved with running water respectively, mix, being made into final concentration is 0.2g/L streptomyces diastatochromogenes D metabolite and 0.01g/L Imidacloprid mixing formula.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104782639A (en) * | 2015-04-23 | 2015-07-22 | 中国计量学院 | Novel method for preventing and controlling nilaparvata lugens filial generation by using fungicide |
| CN104823990A (en) * | 2015-04-23 | 2015-08-12 | 中国计量学院 | Use of fungicide in prevention and control of brown planthopper filial generation |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1187299A (en) * | 1997-01-08 | 1998-07-15 | 张天良 | Efficient pesticide |
| CN101961014A (en) * | 2010-03-19 | 2011-02-02 | 中国计量学院 | Method for preparing microbial pesticide for preventing and controlling fungal diseases of vegetables |
-
2014
- 2014-03-31 CN CN201410128645.3A patent/CN103960288B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1187299A (en) * | 1997-01-08 | 1998-07-15 | 张天良 | Efficient pesticide |
| CN101961014A (en) * | 2010-03-19 | 2011-02-02 | 中国计量学院 | Method for preparing microbial pesticide for preventing and controlling fungal diseases of vegetables |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104782639A (en) * | 2015-04-23 | 2015-07-22 | 中国计量学院 | Novel method for preventing and controlling nilaparvata lugens filial generation by using fungicide |
| CN104823990A (en) * | 2015-04-23 | 2015-08-12 | 中国计量学院 | Use of fungicide in prevention and control of brown planthopper filial generation |
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| CN103960288B (en) | 2015-10-14 |
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