CN103960289B - A kind of preparation method preventing and treating the mixing formula preparation of brown planthopper - Google Patents
A kind of preparation method preventing and treating the mixing formula preparation of brown planthopper Download PDFInfo
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- CN103960289B CN103960289B CN201410128675.4A CN201410128675A CN103960289B CN 103960289 B CN103960289 B CN 103960289B CN 201410128675 A CN201410128675 A CN 201410128675A CN 103960289 B CN103960289 B CN 103960289B
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- brown planthopper
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Abstract
The present invention relates to microbial technology field, particularly relate to a kind of preparation method preventing and treating the mixing formula preparation of brown planthopper.Streptomyces diastatochromogenes (Streptomyces of the present invention? diastatochromogenes), name as streptomyces diastatochromogenes D.This bacterial strain in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, preservation date on May 25th, 2007, preservation registration number CGMCC? No.2060.Streptomyces diastatochromogenes D of the present invention, extracts through fermentation and can obtain the inhibited metabolite of brown planthopper Yeast-like symbionts.The most significant feature of the present invention is that streptomyces diastatochromogenes D metabolite is mixed with into new formula with common insecticides Imidacloprid according to certain ratio, utilization of pesticides amount can be obviously reduced with this mixing formula control brown planthopper, the control efficiency of effective raising brown planthopper, delays the drug-fast development of brown planthopper.
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 (Nilaparvatalugens
) be one of important pests of paddy rice, mainly to cause harm rice plant to sting to inhale phloem sap and propagate the mode of Rice Virus.There will be large-area rice strain withered lodging during the large generation of insect pest, cause grain drop in production, even have no harvest.In production, chemical pesticide is the Main Means of control brown planthopper.But because a large amount of frequent of chemical insecticide uses, brown planthopper creates pesticide resistance to many chemical insecticides, causes control efficiency obviously to decline.Therefore, investigation and application reduce Brown Planthopper occur and the new tool that causes harm extremely urgent.
The symbiosis of insect and microorganism is a kind of ubiquitous phenomenon, and both define inseparable relation during evolution.Insect is that fungal component provides stable microhabitat and nutrition, and insect relies on some the important nutrient component lacked in fungal component purified diet, the metabolic detoxification of fungal component to insect also has certain effect simultaneously, as synthesized important nutrient component for insect, participate in nitrogen cycle, allogenic material etc. in degraded insect bodies.Disappearance fungal component can directly cause insect growth to breed to be obstructed, lethality rising etc.Research shows, also there is multiple Yeast-like symbionts in brown planthopper body in stomach fat body, Parasaccharomyces Ascomycotina (Ascomycotina), gang pyrenomycetes (Pyrenomycetes), and Candida belongs to.Brown planthopper Yeast-like symbionts can be host's synthesis and provides the multiple nutritional components such as essential amino acid, steroid substance and protein, compensate for the imbalance that brown planthopper takes food trophic structure, ensure that normal growth and the breeding of brown planthopper; Yeast-like endosymbiote disappearance can cause brown planthopper growth retardation, arrest of development, even death.Since brown planthopper and Yeast-like symbionts relation in growth and breeding is so tight, controls brown planthopper by the means of " suppress Yeast-like symbionts thus indirectly prevent and treat brown planthopper " and occur and reduce that it causes harm will be a kind of important means of preventing and treating brown planthopper from now on.Because that suppresses that the fungal component in brown planthopper body not only can be direct or indirect reaches the object controlling brown planthopper, but also can effectively improve current insecticide control efficiency, delay the drug-fast development of brown planthopper.
Streptomyces diastatochromogenes D metabolite is mixed with into new formula with common insecticides Imidacloprid according to certain ratio by the present invention, and have studied this formula to the inhibitory action of Yeast-like symbionts in brown planthopper body and the insecticidal effect to brown planthopper.This invention is that the control of Brown Planthopper provides new more efficiently method.
Summary of the invention
The object of the invention is to develop immunity to drugs to conventional chemical pesticides for brown planthopper, cause the deficiency that chemopreventive effects obviously declines, there is provided one to utilize streptomyces diastatochromogenes D metabolite to suppress Yeast-like symbionts growth in brown planthopper body, and then cause the weak even dead new approaches of brown planthopper; Another object of the present invention there is provided a kind of new method and the new medicament of preventing and treating brown planthopper.
Streptomyces diastatochromogenes of the present invention (Streptomycesdiastatochromogenes), names as streptomyces diastatochromogenes D.This bacterial strain in the center preservation of 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 CGMCCNo.2060.Streptomyces diastatochromogenes D of the present invention, extracts through fermentation and can obtain the inhibited metabolite of brown planthopper Yeast-like symbionts.
The object of the invention is achieved through the following technical solutions:
Prevent and treat a mixing formula preparation for brown planthopper, carry out according to the following steps:
(1) activation culture of bacterial classification: moving to being separated the streptomyces diastatochromogenes D bacterial classification preserved in vitro on Gause I inclined-plane, under 28 DEG C ± 1 DEG C condition, being cultured to test tube slant and covering with spore;
(2) fermented and cultured: fermentation culture is containing soybean cake powder 20g, corn flour 15g in every 1000mL, wheat bran 10g, NH
4nO
33g, (NH
4)
2sO
45g, CaCO
35g, MgSO
41g, sterilizing, to be cooled to 40 DEG C, be aseptically seeded in fermentation culture, under 28 DEG C ± 1 DEG C condition, cultivate 5 days;
(3) fermentation product extracts: ferment complete, collected by centrifugation fermented supernatant fluid, obtains medicinal extract with conventional organic solvent extraction after Vacuum Concentration;
(4) drug solution preparing: the medicinal extract obtain step (3) and common insecticides proportioning, for the control of brown planthopper.
Beneficial effect of the present invention: the invention provides a kind of new method 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: moving to being separated the streptomyces diastatochromogenes D bacterial classification preserved in vitro on Gause I inclined-plane, under 28 DEG C ± 1 DEG C condition, being cultured to test tube slant and covering with spore;
(2) fermented and cultured: fermentation culture is containing soybean cake powder 20g, corn flour 15g in every 1000mL, wheat bran 10g, NH
4nO
33g, (NH
4)
2sO
45g, CaCO
35g, MgSO
41g, sterilizing, to be cooled to 40 DEG C, aseptically inoculate a metal platinum filament ring streptomyces diastatochromogenes D spore in fermentation culture, under 28 DEG C ± 1 DEG C condition, cultivate 5 days;
(3) fermentation product extracts: collected by centrifugation zymotic fluid after fermentation, adds equal-volume extraction into ethyl acetate, take off layer, use equal-volume extracting n-butyl alcohol again, get upper strata, be concentrated into medicinal extract under vacuum, 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, mixing, being made into final concentration is 0.2g/L streptomyces diastatochromogenes D metabolite and 0.01g/L Imidacloprid mixing formula.
Embodiment 3:(streptomyces diastatochromogenes D metabolite and the application of Imidacloprid mixing formula in control brown planthopper nymph)
Carry out according to the following steps:
(1) tillering and jointing stage TN1 paddy rice, removes part root and base portion leaf sheath afterwash, naturally dries;
(2) compare with clear water, spraying final concentration is 0.2g/L streptomyces diastatochromogenes D metabolite and 0.01g/L Imidacloprid confection, requires that medicament Pcnten-1 yne-4 is even, coverage rate is high, pharmaceutical quantities is suitable, does not produce loss and is advisable, naturally dry with moistening target surface;
(3) rice seedling after step (2) process puts into the Boiling tube injecting 20ml paddy rice culture fluid in advance, often pipe 3 rice seedlings, access 3 age brown planthopper nymph 30, to spray the rice seedlings process after clear water as negative control, spray 0.04g/L Imidacloprid liquid and be treated to positive control, each process repeats for 3 times;
(4) within 1 day, 2 days and 4 days, measure brown planthopper parachorium bacterium quantity after treatment respectively, and add up brown planthopper lethality.Wherein brown planthopper parachorium bacterium quantitative measurement method is: get the brown planthopper 3 survived in each process respectively, with the alcohol disinfecting of 75%, rinsed with sterile water, remove wing, head and chest, belly is put into 1.5ml centrifuge tube, add the physiological saline 50 μ l of 0.9%, homogenate, then get 2 μ l homogenate drops on blood cell counting plate, close the lid fragmentation, under microscope (16 × 40), add up fungal component quantity.
Embodiment 4:(streptomyces diastatochromogenes D metabolite and the application of Imidacloprid mixing formula in control brown planthopper female adult pest)
Carry out according to the following steps:
(1) tillering and jointing stage TN1 paddy rice, removes part root and base portion leaf sheath afterwash, naturally dries;
(2) compare with clear water, spraying final concentration is 0.2g/L streptomyces diastatochromogenes D metabolite and 0.01g/L Imidacloprid confection, requires that medicament Pcnten-1 yne-4 is even, coverage rate is high, pharmaceutical quantities is suitable, does not produce loss and is advisable, naturally dry with moistening target surface;
(3) rice seedling after step (2) process puts into the Boiling tube injecting 20ml culture fluid in advance, often pipe 4 rice seedlings, the brachypterism female adult pest 30 that access has just been sprouted wings, to spray the rice seedlings process clearly as negative control, spray 0.04g/L Imidacloprid liquid and be treated to positive control, each process repeats for 3 times;
(4) within 1 day, 2 days and 4 days, measure brachypterism female worm brown planthopper parachorium bacterium quantity after treatment respectively, and add up brown planthopper lethality.Wherein brown planthopper parachorium bacterium quantitative measurement method is: get the female worm of the brachypterism of surviving in each process 3 respectively, with the alcohol disinfecting of 75%, rinsed with sterile water, remove wing, head and chest, belly is put into 1.5ml centrifuge tube, add the physiological saline 50 μ l of 0.9%, homogenate, then get 2 μ l homogenate drops on blood cell counting plate, close the lid fragmentation, under microscope (16 × 40), add up 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 inhibit the quantity of brown planthopper parachorium bacterium, thus cause brown planthopper lethality to rise.
Table 1 streptomyces diastatochromogenes D metabolite and Imidacloprid confection are to the control efficiency of brown planthopper nymph
Table 2 streptomyces diastatochromogenes D metabolite and Imidacloprid confection are to the control efficiency of brown planthopper female adult pest
Streptomyces diastatochromogenes D metabolite and the control efficiency of Imidacloprid decrement formula to brown planthopper nymph and female adult pest are better than conventional chemical liquid or similar.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 (1)
1. prevent and treat a preparation method for the mixing formula preparation of brown planthopper, this mixing formula preparation comprise streptomyces diastatochromogenes (
streptomycesdiastatochromogenes) D metabolite and Imidacloprid, described mixing formula preparation is obtained by following methods:
(1) activation culture of bacterial classification: move in vitro on Gause I inclined-plane by being separated the streptomyces diastatochromogenes D bacterial classification preserved, under 28 DEG C ± 1 DEG C condition, be cultured to test tube slant and cover with spore, described streptomyces diastatochromogenes D is in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, and preservation registration number is CGMCCNo.2060;
(2) fermented and cultured: fermentation culture is containing soybean cake powder 20g, corn flour 15g in every 1000mL, wheat bran 10g, NH
4nO
33g, (NH
4)
2sO
45g, CaCO
35g, MgSO
41g, sterilizing, to be cooled to 40 DEG C, aseptically inoculate a metal platinum filament ring streptomyces diastatochromogenes D spore in fermentation culture, under 28 DEG C ± 1 DEG C condition, cultivate 5 days;
(3) fermentation product extracts: ferment complete, collected by centrifugation fermented supernatant fluid, adds equal-volume extraction into ethyl acetate, take off layer, then use equal-volume extracting n-butyl alcohol, get upper strata, be concentrated into medicinal extract under vacuum, this medicinal extract is streptomyces diastatochromogenes D metabolite;
(4) mixing formula preparation: imidacloprid water dispersible granule, active ingredient mass percent is 70%, produced by Beyer Co., Ltd, imidacloprid water dispersible granule and streptomyces diastatochromogenes D metabolite are dissolved with running water respectively, mixing, being made into final concentration is 0.2g/L streptomyces diastatochromogenes D metabolite and 0.01g/L Imidacloprid mixing formula.
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Citations (3)
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| US4056615A (en) * | 1975-05-28 | 1977-11-01 | U C B Societe Anonyme | Lucknomycin and process for producing same |
| CN101225368A (en) * | 2007-11-19 | 2008-07-23 | 中国计量学院 | Biocontrol actinomyces-streptomyces diastatochromogenes D |
| CN102911227A (en) * | 2008-09-27 | 2013-02-06 | 上海市农药研究所 | Streptomyces diastatochromogenes and fermentation product as well as application thereof |
-
2014
- 2014-03-31 CN CN201410128675.4A patent/CN103960289B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4056615A (en) * | 1975-05-28 | 1977-11-01 | U C B Societe Anonyme | Lucknomycin and process for producing same |
| CN101225368A (en) * | 2007-11-19 | 2008-07-23 | 中国计量学院 | Biocontrol actinomyces-streptomyces diastatochromogenes D |
| CN102911227A (en) * | 2008-09-27 | 2013-02-06 | 上海市农药研究所 | Streptomyces diastatochromogenes and fermentation product as well as application thereof |
Non-Patent Citations (5)
| Title |
|---|
| Formation，regeneration，and transformation of protoplasts of Streptomces diastatochromogenes 1628;Zheng ma et al.,;《Folia Microbiologica》;20130731;第59卷;第93-97页 * |
| 共生菌在褐飞虱防治中的应用;陈列忠等;《农药》;20061130;第45卷(第11期);第726-729页 * |
| 常用杀虫剂和杀菌剂对褐飞虱类酵母共生菌生长的影响;陈建明等;《植物保护》;20091231;第35卷(第6期);第47-51页 * |
| 拮抗链霉菌B28的分类鉴定及其生防作用研究;申屠旭萍等;《植物病理学报》;20121231;第42卷(第1期);第105-109页 * |
| 淀粉酶产色链霉菌杀灭钉螺研究;胡代炎等;《中国血吸虫病防治杂志》;19951231;第7卷(第1期);第16-19页 * |
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