CN105385600A - Cotton bollworm microsporidia strain and application of cotton bollworm microsporidia strain in agriculture pest control - Google Patents
Cotton bollworm microsporidia strain and application of cotton bollworm microsporidia strain in agriculture pest control Download PDFInfo
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- CN105385600A CN105385600A CN201510717755.8A CN201510717755A CN105385600A CN 105385600 A CN105385600 A CN 105385600A CN 201510717755 A CN201510717755 A CN 201510717755A CN 105385600 A CN105385600 A CN 105385600A
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Abstract
The invention discloses a cotton bollworm microsporidia strain. The cotton bollworm microsporidia strain FDK001 is preserved in the China General Microbiological Culture Collection Center. The preservation serial number is CGMCC No.11289. A preparation method of the cotton bollworm microsporidia strain includes the following steps of firstly, spraying spore liquid of the cotton bollworm microsporidia strain FDK001 on cotton bollworm feed to feed cotton bollworms; secondly, making cotton bollworms died of infection of cotton bollworm microsporidia after cotton bollworms eat the feed with the cotton bollworm microsporidia strain FDK001; thirdly, taking spores of the cotton bollworm microsporidia strain FDK001 in dead cotton bollworm bodies to serve as active ingredients for agriculture pest control. The cotton bollworm microsporidia strain can be used for controlling various Lepidoptera agriculture pests such as cotton bollworms, plutella xylostella, beet armyworm, cabbage worms, mamestra brassicae, chilo suppressalis and cnaphalocrocis medinalis.
Description
Technical field
The present invention relates to biological control of insect pests technical field, specifically a kind of bollworm microsporidium strain and the application in agricultural insect pests control thereof.
Background technology
Genus heliothis, in lepidopteran Noctuidae, is polyphagous pest-insect.The harm of larva moth food cotton, corn, tomato, the capsicum etc. of bollworm, being China primary pest agriculturally, is also the primary pest on world agriculture.China is since nineteen ninety, and bollworm is great outburst year after year, causes huge financial loss.Afterwards, along with the big area of the Insect Resistant Cotton of trans Bt gene uses, the harm of bollworm in cotton field obtained effective control, but, report that bollworm has created serious resistance to genetically modified Bt albumen in recent years, Brazil in South America, bollworm great outburst once more in recent years.
The method of current control bollworm mainly adopts chemical pesticide control, although chemical prevention can force down the population density of bollworm rapidly, but often cause environmental pollution, kill and wound a large amount of useful animals, person poultry poisoning and insect such as to develop immunity to drugs at the adverse consequences.Biological pesticide control bollworm not only effectively can control the harm of bollworm, and not killed natural enemies, environmentally friendly, to person poultry safety, there is very wide application prospect.Therefore, the effective biological control resource developing other is prevented and treated bollworm, is conducive to improving prevention effect, slows down resistance development, significant.
Microsporidium is entomopathogen, belongs to the entozoic single-celled protozoal of special sexual cell biological.It is once invade in host insect body, just can pass through food, pathogen propagation gives other individual by contact etc., and can be transmitted to offspring by ovum, thus cause the generation of insect microsporidiosis and popular, be one of important factor of nature restriction Insect Population Density.
Although there has been report heliothis armigera nuclear polyhedrosis virus to may be used for biological control bollworm at present, the control of microsporidium to bollworm has had multiple advantage.The first, heliothis armigera nuclear polyhedrosis virus is comparatively harsh to field environment, to ultraviolet-sensitive, and the more viral more UV resistant of microsporidium, the requirement that land for growing field crops uses can be met.The second, single-minded to infect bollworm insecticidal spectrum narrow for heliothis armigera nuclear polyhedrosis virus, and bollworm microsporidium can infect multiple lepidoptera pests such as comprising bollworm, small cabbage moth, beet armyworm, cabbage caterpillar, lopper worm, striped rice borer, Cnaphalocrocis medinali(rice leaf roller), its insecticidal spectrum is wide.Three, heliothis armigera nuclear polyhedrosis virus can only infect bollworm by eating propagation, and vertical transmission can not give the ovum in lower generation.The micro-spore of bollworm vertical transmission can infect the ovum in lower generation, causes follow-on larva with microsporidium, and can infect other insects by contact transmission, control overall worm group radix greatly.
Thus, develop bollworm microsporidium, for agricultural insect pests control, there is realistic meaning.
Summary of the invention
The object of the present invention is to provide a kind of bollworm microsporidium strain, and prevent and treat the application comprised in the multiple lepidoptera pests such as bollworm, small cabbage moth, beet armyworm, cabbage caterpillar, lopper worm, striped rice borer, Cnaphalocrocis medinali(rice leaf roller).
For achieving the above object, the invention provides following technical scheme:
A kind of bollworm microsporidium strain, bollworm microsporidium FDK001 strain, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCCNo.11289.
The preparation method of described bollworm microsporidium strain, comprises the following steps:
(1) spore liquid of bollworm microsporidium FDK001 strain is sprayed on bollworm feed, feeding bollworm;
(2), after bollworm eats the feed with bollworm microsporidium FDK001 strain, to be infected by bollworm microsporidium and dead;
(3) get the spore of the bollworm microsporidium FDK001 strain in dead bollworm polypide, be the activeconstituents of control Agricultural pests.
As the further scheme of the present invention: in step (1), the ratio of described spore liquid and bollworm feed is that 50 microlitre spore liquid are sprayed on the feed of 1 cubic centimetre.
As the present invention's further scheme: the concentration of described spore liquid miospore is 1 × 10
4spore/ml-1 × 10
8spore/ml.
As the present invention's further scheme: the rearing conditions of described bollworm is 25-30 DEG C, relative humidity 60-70%, every day light application time 16 hours.
As the present invention's further scheme: in step (3), specifically can obtain the micro-spore of bollworm microsporidium FDK001 strain described in dead polypide by the following method: by bollworm dead described in step (2) grinding, use 120 object screen filtration again, the tissue juice all filtrations obtained is with after isopyknic water dilution, carry out following differential centrifugation: within centrifugal 15 minutes, get supernatant for 500-1000 rev/min, get precipitation by centrifugal for supernatant liquor 3000 revs/min 30 minutes, be the spore of described FDK001 bollworm microsporidium strain.
The application of described bollworm microsporidium strain in control Agricultural pests.
As the present invention's further scheme: described insect is lepidoptera pest, comprise bollworm, small cabbage moth, beet armyworm, cabbage caterpillar, lopper worm, striped rice borer and Cnaphalocrocis medinali(rice leaf roller).
As the present invention's further scheme: comprise the agricultural insect pests control agent containing described bollworm microsporidium FDK001 strain spore components.
As the present invention's further scheme: described control agent comprises wetting agent, for tween 80, the mixture of one or more in Sodium dodecylbenzene sulfonate, sodium lauryl sulphate, sodium alkyl naphthalene sulfonate, fatty alcohol polyglycol ether vitriol, fatty alcohol-polyoxyethylene ether, polyoxyethylene nonylphenol ether, Span Soxylat A 25-7; Comprising dispersion agent, is one or more mixtures in NNO, sodium lignosulfonate, calcium lignin sulphonate, sodium polyacrylate, methylene-bis naphthalenesulfonate, alkylamino taurate, sodium methylene bis-naphthalene sulfonate formaldehyde condensation products, polyoxyethylene polyoxypropylene block copolymer, carboxymethyl cellulose, polyvinyl alcohol, polyvinylpyrrolidone, tripoly phosphate sodium STPP, sulfonated alkyl naphathalene etc.; Comprising frostproofer, is the mixture of one or more in sucrose, ethylene glycol, propylene glycol, glycerine etc.; Comprising ultraviolet protective agent, is the mixture of one or more in phenylalanine, tyrosine, tryptophane, benzophenone, dimethoxy-benzophenone, octyl methoxycinnamate, octyl salicylate etc.
Compared with prior art, the invention has the beneficial effects as follows: bollworm microsporidium provided by the invention may be used for the multiple lepidopteran Agricultural pests such as control bollworm, small cabbage moth, beet armyworm, cabbage caterpillar, lopper worm, striped rice borer, Cnaphalocrocis medinali(rice leaf roller).This product belongs to biological pesticide, to environment and people, animal, fowl toxicological harmless, to natural enemies security, has a extensive future.
Embodiment
Following examples of implementation are convenient to better understand the present invention, but do not limit the present invention.Experimental technique in following examples of implementation, if no special instructions, is ordinary method.Experiment material used in following examples of implementation, if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative experiment in following examples, all arranges and repeats experiment for three times, results averaged.
Embodiment 1
The acquisition of bollworm microsporidium FDK001 strain
(the dead worm of bollworm mostly is black death, and do not change water, polypide is soft field to be gathered ill bollworm, epidermis easily breaks, and it is thicker that middle phleboedesis is swollen, fragile easily broken), grinding, filtration, microscopy, microsporidium is colourless under the microscope, and refractivity is strong, long avette, minority is kidney shape, and size is about 4.5um × 2.5um, dyes with Giemsa, core is colored as light red, and tenuigenin dyeing is for blue.Each wild cotton earworm microsporidium strain is spread upon on bollworm feed with a series of concentration, bollworm of feeding afterwards, the dead polypide (representing the strain that virulence is strong) comparatively early of results, through grinding, filter, centrifugal etc.Obtain the bollworm microsporidium of purifying.Step sizing 10 generation, obtain bollworm microsporidium (
nosemaheliothidis) FDK001 strain.
On October 19th, 2015 by bollworm microsporidium (
nosemaheliothidis) FDK001 strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (be called for short CGMCC, No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, address), deposit number CGMCCNo.11289.Bollworm microsporidium (
nosemaheliothidis) FDK001 strain CGMCCNo.11289 abbreviation FDK001 strain.
Embodiment 2
The pathogenic qualification of FDK001 strain
Bollworm microsporidium FDK001 strain blood counting chamber determination concentration.The infection experiment of bollworm microsporidium FDK001 strain to bollworm, small cabbage moth, beet armyworm, cabbage caterpillar, lopper worm, striped rice borer, Cnaphalocrocis medinali(rice leaf roller) adopts micro-spore FDK001 strain suspension (10
6spore/ml) method of Direct spraying on ovum grain.To the infection experiment of bollworm, small cabbage moth, beet armyworm, cabbage caterpillar, lopper worm, striped rice borer, Cnaphalocrocis medinali(rice leaf roller) 2 instar larvae, by micro-spore FDK001 strain suspension (10
6spore/ml) 50 microlitres are sprayed on the artificial diet of 1 cubic centimetre, in 26 DEG C, relative humidity 60%, every day light application time 16 hours/day.Infect and raise.Record larval mortality.Each process repeats for 3 times, each repetition 360 cephalonts.
Arrange blank group, the mortality ratio of blank group larva is efficiency test below 1% simultaneously.
What pathogenic infection was tested the results are shown in Table 1.
Pathogenic infection result shows that bollworm microsporidium FDK001 strain all has strong pathogenic infectious to bollworm, small cabbage moth, beet armyworm, cabbage caterpillar, lopper worm, striped rice borer, Cnaphalocrocis medinali(rice leaf roller).
Table 1 bollworm microsporidium FDK001 strain 10
6spore/ml is to the infection experiment of several insect
Examples of implementation 3
The preparation of bollworm microsporidium FDK001 strain spore
The preparation of bollworm microsporidium FDK001 strain spore, comprises the steps:
(1) by the spore liquid concentration 1 × 10 of bollworm microsporidium FDK001 strain
6spore/ml is sprayed at (50 microlitre spore liquid are sprayed on the feed of 1 cubic centimetre) on bollworm feed, feeding bollworm;
(2) after bollworm eats the feed with bollworm microsporidium FDK001 strain, bollworm is dead by the infection of bollworm microsporidium FDK001 strain, the rearing conditions of described bollworm is 25-30 DEG C, relative humidity 60-70%, every day light application time 16 hours;
(3) bollworm dead described in step (2) is ground, uses 120 object screen filtration again.The tissue juice all filtrations obtained is with after isopyknic water (being preferably distilled water) dilution, carry out following differential centrifugation: within centrifugal 15 minutes, get supernatant for 500-1000 rev/min, get precipitation by centrifugal for supernatant liquor 3000 revs/min 30 minutes, be the spore of described FDK001 bollworm microsporidium strain.Be separated the spore of the bollworm microsporidium FDK001 strain in the bollworm polypide of the death obtained, be the activeconstituents of control Agricultural pests.
Examples of implementation 4
The Synthesis and applications of bollworm microsporidium FDK001 strain spore preparation
By the bollworm microsporidium FDK001 strain spore obtained in examples of implementation 3, add 5% wetting agent tween 80,5% dispersion agent sodium lignosulfonate, 5% frostproofer sucrose, 1% ultraviolet protective agent tryptophane is prepared as suspension agent.Wherein the content of bollworm microsporidium FDK001 strain spore is 2 × 10
9spore/ml.
Above-mentioned suspension agent is watered 500 times bollworm in a 2 Sheng phase in ages and carries out land for growing field crops spraying, every mu uses 100 ml of formulation.Arrange 3 to repeat and blank group.Spray and within latter 7 days, add up the bollworm quantity in the experiment land for growing field crops of carrying out spraying, calculate preventive effect.Microsporidium FDK001 strain spore preparation preventive effect is 81%.
Above the better embodiment of this patent is explained in detail, but this patent is not limited to above-mentioned embodiment, in the ken that one skilled in the relevant art possesses, can also makes a variety of changes under the prerequisite not departing from this patent aim.
Claims (10)
1. a bollworm microsporidium strain, is characterized in that, bollworm microsporidium FDK001 strain, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCCNo.11289.
2. a preparation method for bollworm microsporidium strain as claimed in claim 1, is characterized in that, comprise the following steps:
(1) spore liquid of bollworm microsporidium FDK001 strain is sprayed on bollworm feed, feeding bollworm;
(2), after bollworm eats the feed with bollworm microsporidium FDK001 strain, to be infected by bollworm microsporidium and dead;
(3) get the spore of the bollworm microsporidium FDK001 strain in dead bollworm polypide, be the activeconstituents of control Agricultural pests.
3. the preparation method of bollworm microsporidium strain according to claim 2, is characterized in that, in step (1), the ratio of described spore liquid and bollworm feed is that 50 microlitre spore liquid are sprayed on the feed of 1 cubic centimetre.
4. the preparation method of bollworm microsporidium strain according to claim 2, is characterized in that, the concentration of described spore liquid miospore is 1 × 10
4spore/ml-1 × 10
8spore/ml.
5. the preparation method of bollworm microsporidium strain according to claim 2, is characterized in that, the rearing conditions of described bollworm is 25-30 DEG C, relative humidity 60-70%, every day light application time 16 hours.
6. the preparation method of bollworm microsporidium strain according to claim 2, it is characterized in that, in step (3), specifically can obtain the micro-spore of bollworm microsporidium FDK001 strain described in dead polypide by the following method: by bollworm dead described in step (2) grinding, use 120 object screen filtration again, the tissue juice all filtrations obtained is with after isopyknic water dilution, carry out following differential centrifugation: within centrifugal 15 minutes, get supernatant for 500-1000 rev/min, precipitation is got by centrifugal for supernatant liquor 3000 revs/min 30 minutes, be the spore of described FDK001 bollworm microsporidium strain.
7. the application of bollworm microsporidium strain according to claim 1 in control Agricultural pests.
8. the application of bollworm microsporidium strain according to claim 7 in control Agricultural pests, it is characterized in that, described insect is lepidoptera pest, comprises bollworm, small cabbage moth, beet armyworm, cabbage caterpillar, lopper worm, striped rice borer and Cnaphalocrocis medinali(rice leaf roller).
9. the application of bollworm microsporidium strain according to claim 7 in control Agricultural pests, is characterized in that, comprise the agricultural insect pests control agent containing described bollworm microsporidium FDK001 strain spore components.
10. the application of bollworm microsporidium strain according to claim 9 in control Agricultural pests, it is characterized in that, described control agent comprises wetting agent, for tween 80, the mixture of one or more in Sodium dodecylbenzene sulfonate, sodium lauryl sulphate, sodium alkyl naphthalene sulfonate, fatty alcohol polyglycol ether vitriol, fatty alcohol-polyoxyethylene ether, polyoxyethylene nonylphenol ether, Span Soxylat A 25-7; Comprising dispersion agent, is one or more mixtures in NNO, sodium lignosulfonate, calcium lignin sulphonate, sodium polyacrylate, methylene-bis naphthalenesulfonate, alkylamino taurate, sodium methylene bis-naphthalene sulfonate formaldehyde condensation products, polyoxyethylene polyoxypropylene block copolymer, carboxymethyl cellulose, polyvinyl alcohol, polyvinylpyrrolidone, tripoly phosphate sodium STPP, sulfonated alkyl naphathalene etc.; Comprising frostproofer, is the mixture of one or more in sucrose, ethylene glycol, propylene glycol, glycerine etc.; Comprising ultraviolet protective agent, is the mixture of one or more in phenylalanine, tyrosine, tryptophane, benzophenone, dimethoxy-benzophenone, octyl methoxycinnamate, octyl salicylate etc.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115147837A (en) * | 2022-08-16 | 2022-10-04 | 河北省农林科学院植物保护研究所 | Athetis lepigone breeding method and system based on optical image recognition |
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| CN104232487A (en) * | 2014-08-27 | 2014-12-24 | 河北省农林科学院植物保护研究所 | Nosema bombycis for controlling Athetis lepigone and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104232487A (en) * | 2014-08-27 | 2014-12-24 | 河北省农林科学院植物保护研究所 | Nosema bombycis for controlling Athetis lepigone and application thereof |
Non-Patent Citations (2)
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| 冯书亮等: "棉铃虫微孢子虫疾病流行对其发生危害的控制作用", 《全国生物防治学术研讨会》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115147837A (en) * | 2022-08-16 | 2022-10-04 | 河北省农林科学院植物保护研究所 | Athetis lepigone breeding method and system based on optical image recognition |
| CN115147837B (en) * | 2022-08-16 | 2023-10-27 | 河北省农林科学院植物保护研究所 | Athetis lepigone feeding method and system based on optical image recognition |
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