CN103931665A - Mixed formulation preparation for prevention and control of nilaparvata lugens - Google Patents
Mixed formulation preparation for prevention and control of nilaparvata lugens Download PDFInfo
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- CN103931665A CN103931665A CN201410191332.2A CN201410191332A CN103931665A CN 103931665 A CN103931665 A CN 103931665A CN 201410191332 A CN201410191332 A CN 201410191332A CN 103931665 A CN103931665 A CN 103931665A
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Abstract
The invention relates to the technical field of biological prevention and control, and particularly relates to a (i) Trichodermabrevicompactum (/i) 0248 metabolite and imidacloprid mixed formulation preparation. The Trichodermabrevicompactum disclosed by the invention is named Trichodermabrevicompactum 0248, and the strain with a Preservation No. of CGMCCNo.6985 is preserved in China General Microbiological Culture Collection Center (CGMCC) on December 12, 2012. Through carrying out fermenting and extracting on the Trichodermabrevicompactum 0248 disclosed by the invention, metabolites having an inhibiting effect on nilaparvata lugens-like yeast symbiotes can be obtained. The mixed formulation preparation disclosed by the invention has the most obvious characteristics that metabolites of Trichodermabrevicompactum 0248 and a conventional pesticide imidacloprid are mixed in certain proportion so as to prepare a new formulation, and in the case of preventing and controlling nilaparvata lugens by using the mixed formulation, the application amount of pesticides can be significantly reduced, the prevention and control effect on nilaparvata lugens can be effectively improved, and the development of insecticide resistance of nilaparvata lugens can be delayed.
Description
Technical field
The present invention relates to biological prevention field, particularly relate to the application of navel spore Trichoderma 0248 metabolite and Imidacloprid mixing formula preparation.
Background technology
Brown planthopper (
nilaparvata lugensst is l) one of the important pests of paddy rice, the mode of mainly inhaling phloem sap with thorn and the propagating Rice Virus rice plant of causing harm.When 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), gang pyrenomycetes (
pyrenomycetes),
candidabelong 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 brown planthopper and take food the imbalance of trophic structure, ensured 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, controlling by the means of " indirectly preventing and treating brown planthopper thereby suppress class yeast fungal component " 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.
Summary of the invention
The present invention by navel spore Trichoderma (
trichoderma brevicompactum) 0248 metabolite is mixed with into new formula with common insecticides Imidacloprid according to certain ratio, and studied the inhibitory action of this formula to class yeast fungal component in brown planthopper body and the insecticidal effect to brown planthopper.Effective control that this invention is Brown Planthopper provides new method.
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 one to utilize navel spore Trichoderma 0248 metabolite to suppress class yeast fungal component growth in brown planthopper body, and then cause the new approaches of brown planthopper death; Another object of the present invention has been to provide a kind of new method and new medicament of preventing and treating brown planthopper.
Navel spore Trichoderma of the present invention (
trichoderma brevicompactum), name as navel spore Trichoderma 0248.This bacterial strain is 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 December 12nd, 2012, preservation registration number CGMCC No. 6985.
Navel spore Trichoderma 0248 of the present invention, 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:
The application in control brown planthopper of navel spore Trichoderma 0248 metabolite and Imidacloprid mixing formula preparation, carry out according to the following steps:
(1) activation culture of bacterial classification: move on potato glucose medium (PDA) flat board separating navel spore Trichoderma 0248 bacterial classification of preserving, under 28 DEG C ± 1 DEG C condition, be cultured to culture dish and cover with green spores, add 10ml sterile water, scrape gently with spreading rod, spore liquid under washing is filtered with aseptic filter paper, obtains spore suspension, and to make spore suspension concentration with haemocyte technology plate counting be 1.0 × 10
6individual/ml;
(2) fermented and cultured: fermentation culture is to contain glucose 8g in every 1000ml, beef extract 10g, NaCl 2g, the bottled fermentation culture 60ml of every 300ml triangle; 121 DEG C of sterilizing 20min immediately after preparation, to be cooled to 40 DEG C under aseptic condition inoculum density be 1.0 × 10
6the spore suspension 1ml of individual/ml, under 28 DEG C ± 2 DEG C conditions, cultivates 3 days;
(3) fermentating metabolism product is collected: ferment complete, and centrifugal, abandon mycelium, collect zymotic fluid, this zymotic fluid is navel spore Trichoderma 0248 metabolite;
(4) liquid preparation: navel spore Trichoderma 0248 metabolite and common insecticides proportioning that step (4) is obtained, for the control of brown planthopper.
Beneficial effect of the present invention: the invention provides a kind of new method of utilizing navel spore Trichoderma 0248 metabolite and Imidacloprid mixing formula control brown planthopper.
Embodiment
The preparation of embodiment 1:(navel spore Trichoderma 0248 metabolite)
Carry out according to the following steps:
(1) activation culture of bacterial classification: move on potato glucose medium (PDA) flat board separating navel spore Trichoderma 0248 bacterial classification of preserving, under 28 DEG C ± 1 DEG C condition, cultivate 96h, cover with green spores to culture dish, add 10ml sterile water, scrape gently with spreading rod, the spore liquid under washing is filtered with aseptic filter paper, obtain without mycelium spore suspension, and to make spore suspension concentration with haemocyte technology plate counting be 1.0 × 10
6individual/ml;
(3) fermented and cultured: fermentation culture is to contain glucose 8g in every 1000ml, beef extract 10g, NaCl 2g, the bottled fermentation culture 60ml of every 300ml triangle; 121 DEG C of sterilizing 20min immediately after preparation, to be cooled to 40 DEG C under aseptic condition inoculum density be 1.0 × 10
6the spore suspension 1ml of individual/ml, under 28 DEG C ± 2 DEG C conditions, cultivates 3 days;
(4) fermentation product metabolite is collected: ferment complete, collect zymotic fluid, this zymotic fluid is navel spore Trichoderma 0248 metabolite;
The preparation of embodiment 2:(navel spore Trichoderma 0248 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 is dissolved with running water, being made into concentration is 0.01g/L Imidacloprid liquid, then mix with navel spore Trichoderma 0248 metabolite equal-volume, be prepared into Imidacloprid and navel spore Trichoderma 0248 metabolite mixing formula (in following content, claiming " product A ").Concentration is that 0.01g/L Imidacloprid liquid mixes with 1:2 volume ratio with navel spore Trichoderma 0248 metabolite, is prepared into another kind of Imidacloprid and navel spore Trichoderma 0248 metabolite mixing formula (in following content, claiming " product B ").
Embodiment 3:(navel spore Trichoderma 0248 metabolite and the Imidacloprid mixing formula application in control brown planthopper nymph)
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) compare with clear water, spray respectively " product A " and " product B " prepared by embodiment 2, require liquid droplet to be evenly distributed, coverage rate is high, and amount of liquid medicine is suitable, does not produce to run off be advisable with moistening object surface, naturally dries;
(3) put into the Boiling tube of prior injection 20ml paddy rice culture fluid through step (2) rice seedling after treatment, 3 rice seedlings of every pipe, access 3 age 30 of brown planthopper nymphs, process as negative control using the rice seedlings spraying after clear water, spray 0.04g/L Imidacloprid liquid and be treated to positive control, each processing repeats for 3 times;
(4) respectively in processing latter 1 day, within 2 days and 4 days, measure brown planthopper parachorium bacterium quantity, and add up brown planthopper lethality.Wherein brown planthopper parachorium bacterium quantitative measurement method is: get respectively 3 of the brown planthoppers that survive in each processing, with 75% alcohol disinfecting, 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.
Embodiment 4:(navel spore Trichoderma 0248 metabolite and the Imidacloprid mixing formula application in the female adult of control brown planthopper)
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) compare with clear water, spray respectively " product A " and " product B " prepared by embodiment 2,, require liquid droplet to be evenly distributed, coverage rate is high, and amount of liquid medicine is suitable, does not produce to run off be advisable with moistening object surface, naturally dries;
(3) put into the Boiling tube of prior injection 20ml paddy rice culture fluid through step (2) rice seedling after treatment, 4 rice seedlings of every pipe, 30 of the female adults of brachypterism that access has just been sprouted wings, process as negative control using the rice seedlings spraying after clear water, spray 0.08g/L Imidacloprid liquid and be treated to positive control, each processing repeats for 3 times;
(4) respectively in processing latter 1 day, within 2 days and 4 days, measure the female worm brown planthopper of brachypterism parachorium bacterium quantity, and add up brown planthopper lethality.Wherein brown planthopper parachorium bacterium quantitative measurement method is: get respectively 3 of the female worms of the brachypterism of surviving in each processing, with 75% alcohol disinfecting, 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: (product of the present invention and conventional pesticide control efficiency comparative trial)
Result of the test is in table 1 and table 2.As seen from Table 1 and Table 2, utilize product A and product B all obviously to suppress the quantity of brown planthopper parachorium bacterium, thereby cause that brown planthopper lethality rises.
Table 1 navel spore Trichoderma 0248 metabolite and the control efficiency of Imidacloprid admixing medical solutions to brown planthopper nymph
Table 2 navel spore Trichoderma 0248 metabolite and the control efficiency of Imidacloprid admixing medical solutions to the female adult of brown planthopper
Navel spore Trichoderma 0248 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 navel spore Trichoderma of the present invention 0248 metabolite 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 (4)
1. prevent and treat a mixing formula preparation for brown planthopper, this mixing formula preparation comprises navel spore Trichoderma 0248 metabolite and Imidacloprid.
2. mixing formula preparation claimed in claim 1, wherein said navel spore Trichoderma 0248 metabolite is made by strain activation and culture, fermented and cultured, fermentating metabolism product collection method.
3. mixing formula preparation claimed in claim 1, wherein said navel spore Trichoderma 0248 metabolite and the volume ratio of 0.01g/L Imidacloprid liquid are 1:1-2:1.
4. mixing formula preparation claimed in claim 1, is obtained by following methods:
(1) activation culture of bacterial classification: move on potato glucose medium (PDA) flat board separating navel spore Trichoderma 0248 bacterial classification of preserving, under 28 DEG C ± 1 DEG C condition, activation culture to culture dish covers with green spores, add 10ml sterile water, scrape gently with spreading rod, spore liquid under washing is filtered with aseptic filter paper, obtains spore suspension, and to make spore suspension concentration with haemocyte technology plate counting be 1.0 × 10
6individual/ml, described navel spore Trichoderma 0248 is in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, preservation date on December 12nd, 2012, preservation registration number CGMCC No. 6985;
(2) fermented and cultured: fermentation culture is to contain glucose 8g in every 1000ml, beef extract 10g, NaCl 2g, the bottled fermentation culture 60ml of every 300ml triangle; 121 DEG C of sterilizing 20min immediately after preparation, to be cooled to 40 DEG C under aseptic condition inoculum density be 1.0 × 10
6the spore suspension 1ml of individual/ml, under 28 DEG C ± 2 DEG C conditions, cultivates 3 days;
(3) fermentating metabolism product is collected: ferment complete, and centrifugal, abandon mycelium, collect zymotic fluid, this zymotic fluid is navel spore Trichoderma 0248 metabolite;
(4) liquid preparation: navel spore Trichoderma 0248 metabolite and common insecticides proportioning that step (4) is obtained, for the control of brown planthopper.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104342461A (en) * | 2014-10-15 | 2015-02-11 | 中国计量学院 | Method for improving synthesis of trichodermin from trichoderma brevicompactum |
| CN104357471A (en) * | 2014-10-15 | 2015-02-18 | 中国计量学院 | Application of gene knock-out carrier in gene function research of trichoderma brevicompactum |
| CN110269067A (en) * | 2018-03-16 | 2019-09-24 | 成都特普生物科技股份有限公司 | A kind of prevention and treatment agricultural has the combination preparation of pest type |
-
2014
- 2014-05-08 CN CN201410191332.2A patent/CN103931665B/en active Active
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104342461A (en) * | 2014-10-15 | 2015-02-11 | 中国计量学院 | Method for improving synthesis of trichodermin from trichoderma brevicompactum |
| CN104357471A (en) * | 2014-10-15 | 2015-02-18 | 中国计量学院 | Application of gene knock-out carrier in gene function research of trichoderma brevicompactum |
| CN110269067A (en) * | 2018-03-16 | 2019-09-24 | 成都特普生物科技股份有限公司 | A kind of prevention and treatment agricultural has the combination preparation of pest type |
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| CN103931665B (en) | 2015-10-07 |
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Address after: Hangzhou City, Zhejiang province 310018 Xiasha Higher Education Park source Street No. 258 Patentee after: CHINA JILIANG UNIVERSITY Address before: 310018 School of life science, No. 258, Xiasha High Education Park, Hangzhou, Zhejiang Province, School of life science, China Jiliang University Patentee before: China Jiliang University |