CN103975957B - A kind of mixing formula formulation preparation method preventing and treating brown planthopper - Google Patents
A kind of mixing formula formulation preparation method preventing and treating brown planthopper Download PDFInfo
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Abstract
The present invention relates to technical field of biological control, particularly relate to a hilum spore Trichoderma (<i>Trichoderma? brevicompactum</i>) 0248 metabolite and Imidacloprid mixing formula preparation.Navel spore Trichoderma of the present invention, names as navel spore Trichoderma 0248.This bacterial strain 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.Navel spore Trichoderma 0248 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 navel spore Trichoderma 0248 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 technical field of biological control, particularly relate to the application of navel spore Trichoderma 0248 metabolite and Imidacloprid mixing formula preparation.
Background technology
Brown planthopper (
nilaparvatalugensst l) 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),
candidabelong 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.
Summary of the invention
The present invention by navel spore Trichoderma (
trichodermabrevicompactum) 0248 metabolite is mixed with into new formula with common insecticides Imidacloprid according to certain ratio, 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 effective control of Brown Planthopper provides new method.
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 navel spore Trichoderma 0248 metabolite to suppress Yeast-like symbionts growth in brown planthopper body, and then cause the new approaches of brown planthopper death; Another object of the present invention there is provided a kind of new method and the new medicament of preventing and treating brown planthopper.
Navel spore Trichoderma of the present invention (
trichodermabrevicompactum), name as navel spore Trichoderma 0248.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 December 12nd, 2012, preservation registration number CGMCCNo.6985.
Navel spore Trichoderma 0248 of the present invention, can obtain the inhibited metabolite of brown planthopper Yeast-like symbionts through fermentation.
The object of the invention is achieved through the following technical solutions:
Navel spore Trichoderma 0248 metabolite and the application of Imidacloprid mixing formula preparation in control brown planthopper, carry out according to the following steps:
(1) activation culture of bacterial classification: move on potato dextrose medium (PDA) flat board by being separated navel spore Trichoderma 0248 bacterial classification preserved, 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 aseptic filter paper under washing filters, and obtains spore suspension, and makes spore suspension concentration be 1.0 × 10 with haemocyte technology plate counting
6individual/ml;
(2) fermented and cultured: fermentation culture is containing glucose 8g in every 1000ml, beef extract 10g, NaCl2g, 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 aseptically 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, centrifugal, abandons mycelium, and collect zymotic fluid, this zymotic fluid is navel spore Trichoderma 0248 metabolite;
(4) drug solution preparing: navel spore Trichoderma 0248 metabolite step (4) obtained 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 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 dextrose medium (PDA) flat board by being separated navel spore Trichoderma 0248 bacterial classification preserved, under 28 DEG C ± 1 DEG C condition, cultivate 96h, green spores is covered with to culture dish, add 10ml sterile water, scrape gently with spreading rod, the spore liquid aseptic filter paper under washing filters, obtain without mycelium spore suspension, and make spore suspension concentration be 1.0 × 10 with haemocyte technology plate counting
6individual/ml;
(3) fermented and cultured: fermentation culture is containing glucose 8g in every 1000ml, beef extract 10g, NaCl2g, 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 aseptically 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, and 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 running water is dissolved, 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 (claiming " product A " in following content).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 (claiming " product B " in following content).
Embodiment 3:(navel spore Trichoderma 0248 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, spray respectively " product A " and " product B " prepared by embodiment 2, require that liquid Pcnten-1 yne-4 is even, coverage rate is high, and amount of liquid medicine 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) difference 1 day after treatment, measures brown planthopper parachorium bacterium quantity for 2 days and 4 days, and adds 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:(navel spore Trichoderma 0248 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, spray respectively " product A " and " product B " prepared by embodiment 2, require that liquid Pcnten-1 yne-4 is even, coverage rate is high, and amount of liquid medicine 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 4 rice seedlings, the brachypterism female adult pest 30 that access has just been sprouted wings, to spray the rice seedlings process after clear water as negative control, spray 0.08g/L Imidacloprid liquid and be treated to positive control, each process repeats for 3 times;
(4) difference 1 day after treatment, measures brachypterism female worm brown planthopper parachorium bacterium quantity for 2 days and 4 days, and adds 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: (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 inhibit the quantity of brown planthopper parachorium bacterium, thus cause brown planthopper lethality to rise.
Table 1 navel spore Trichoderma 0248 metabolite and Imidacloprid admixing medical solutions are to the control efficiency of brown planthopper nymph
Table 2 navel spore Trichoderma 0248 metabolite and Imidacloprid admixing medical solutions are to the control efficiency of brown planthopper female adult pest
Navel spore Trichoderma 0248 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 navel spore Trichoderma 0248 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 mixing formula formulation preparation method for brown planthopper, this mixing formula preparation comprise navel spore Trichoderma (
trichodermabrevicompactum) 0248 metabolite and Imidacloprid liquid, the volume ratio of described navel spore Trichoderma 0248 metabolite and 0.01g/L Imidacloprid liquid is 1:1-2:1; Described mixing formula preparation is obtained by following methods:
(1) activation culture of bacterial classification: move on potato dextrose medium flat board by being separated navel spore Trichoderma 0248 bacterial classification preserved, 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 aseptic filter paper under washing filters, and obtains spore suspension, and makes spore suspension concentration be 1.0 × 10 with blood cell counting plate counting
6individual/ml; Described navel spore Trichoderma 0248 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 CGMCCNo.6985;
(2) fermented and cultured: fermentation culture is containing glucose 8g in every 1000ml, beef extract 10g, NaCl2g, 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 aseptically 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, centrifugal, abandons mycelium, and collect zymotic fluid, this zymotic fluid is navel spore Trichoderma 0248 metabolite;
(4) mixing formula preparation: navel spore Trichoderma 0248 metabolite and 0.01g/L Imidacloprid liquid are pressed the mixing of 1:1-2:1 volume ratio, obtain mixing formula, prevent and treat for brown planthopper.
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| CN104342461A (en) * | 2014-10-15 | 2015-02-11 | 中国计量学院 | Method for improving synthesis of trichodermin from trichoderma brevicompactum |
| CN112273390B (en) * | 2020-06-30 | 2021-12-10 | 中国农业科学院生物技术研究所 | Herbicidal activity of compounds harzianums and producing bacteria thereof |
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| 脐孢木霉菌Trichoderma brevicompactum 0248 tri5基因片段的克隆及表达分析;詹晓奂等;《中国生物防治学报》;20131130;第29卷(第4期);第555-560页 * |
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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, China Jiliang University, Xiasha High Education Park, Hangzhou, Zhejiang Patentee before: China Jiliang University |