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Clanis bilineata tsingtauica cytoplasmic polyhedrosis virus strain as well as proliferation method and application thereof

A technology for cytoplasmic polyhedrosis and bean hawkmoth, applied in the field of microorganisms, can solve problems such as easy mixing of large impurities, inconvenience in collecting insect larvae corpses, and no research or application report on bean hawkmoth plasmic polyhedrosis virus, and avoid the problems of Pollution, the effect of strong insecticidal power

Active Publication Date: 2021-06-15
INST OF MICROBIOLOGY JIANGXI ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, so far, there is no related research or application report
[0006] The use of new bioinsecticides for hordea polyhedrosis virus will not only effectively overcome the problems of drug resistance and environmental pollution caused by the use of chemical pesticides to control bean hordea, but will also overcome the problem of collecting insects in the production of nuclear polyhedrosis virus. Disadvantages of inconvenient larval carcasses and easy mixing of large amounts of impurities

Method used

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  • Clanis bilineata tsingtauica cytoplasmic polyhedrosis virus strain as well as proliferation method and application thereof
  • Clanis bilineata tsingtauica cytoplasmic polyhedrosis virus strain as well as proliferation method and application thereof
  • Clanis bilineata tsingtauica cytoplasmic polyhedrosis virus strain as well as proliferation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: Separation and purification method of bean hornworm plastotype polyhedrosis virus

[0019] The virus of the present invention is isolated from the dead samples of bean hornworm larvae that died naturally from the disease in soybean planting fields in Hubei. The epidermis of the insect corpse is not liquefied and shriveled. The corpse has no obvious signs of fungal fruiting body growth, and the corpse has no foul smell. It presents a typical phenotype of cytoplasmic polyhedrosis virus infection and death. The phenotype of the corpse is as follows: figure 1 shown.

[0020] After the insect corpses are collected, put them into sterile 50mL plastic tubes and bring them back to the laboratory for identification. Use a high-pressure sterilized mortar to grind the insect corpses, add 0.1wt% SDS solution, mix well, pour into a funnel with four layers of gauze and filter to remove the tissue residues of the insect corpses, and use a 50mL plastic tube to collect an...

Embodiment 2

[0022] Example 2: Electron Microscopic Observation of the Microscopic Morphology of Bean Hornmoth Phytotype Polyhedrosis Virus

[0023] Scanning electron microscope sample preparation: take the purified polyhedron precipitate and mix it with a small amount of distilled water, negatively stain with 2wt% phosphotungstic acid with a pH of 10.6 for 2-3min, add it dropwise on the 400-mesh copper mesh Fomnvar membrane, and filter paper from the edge of the copper mesh Gently suck off the excess negative staining solution; electron microscope section preparation: take the purified polyhedron precipitate and add 2.5wt% glutaraldehyde-0.1mol / L phosphate buffer solution with a pH of 7.2, and fix it in a refrigerator at 4°C; Wash three times with 0.1mol / L PBS, 15min each time; then fix with 1wt% osmic acid at room temperature for 2-3h; wash three times with 0.1mol / L PBS with pH 7.2; , 100wt% series concentration of ethanol for dehydration; soak in acetone for 15 minutes, then soak in ace...

Embodiment 3

[0025] Example 3: Secondary Infection of Scutellaridae Larvae and Phenotypic Analysis

[0026] After initially confirming that the pathogen is a cytoplasmic polyhedrosis virus through phenotypic analysis and microscopic examination, in order to further verify that the pathogen is indeed our purified cytoplasmic polyhedrosis virus particles, we smeared these purified aqueous solution of polyhedron particles on fresh Phyllostachys japonica Peach leaves, the leaves were fed to healthy oleander hawkmoth larvae belonging to the family Sculptidae, and it was found that the grinding liquid of insect corpses could infect and kill oleander hawkmoth larvae 100% (N=120); in addition, we found that , the phenotypes of these oleander larvae after death were very similar to those of the dead larvae of the oleander larvae collected in the field: that is, the epidermis was intact, without liquefaction symptoms, shriveled, and the carcasses had no foul smell, and Zombie-like phenotype after no...

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Abstract

The invention discloses a clanis bilineata tsingtauica cytoplasmic polyhedrosis virus strain and a proliferation method thereof. The virus strain is clanis bilineata tsingtauica cytoplasmic polyhedrosis virus, and the preservation name is clanis bilineata tsingtauica cytoplasmic polyhedrosis virus CbCPV. The clanis bilineata tsingtauica cytoplasmic polyhedrosis virus can effectively control clanis bilineata tsingtauica harm, continuously acts after one-time application, can maintain no clanis bilineata tsingtauica harm for two to three years, is convenient to produce, reduces the use of chemical pesticides, is environment-friendly, and can greatly reduce the clanis bilineata tsingtauica harm in soybean production, which assists the work of grain battling with insects in soybean production in China.

Description

technical field [0001] The invention relates to the technical field of microbes, in particular to a strain of the hordea polyhedrosis virus strain and its propagation method and application. Background technique [0002] Bean hornworm, also known as soybean hornworm, is mainly distributed in the Huanghuai River Basin, Yangtze River basin and South China. Its larvae are commonly known as bean dan, bean worm, and bean cicada. , black locust, acacia, Lidou and other plants, one or two generations a year, with mature larvae overwintering in the soil as a soil chamber, pupation and emergence in June of the following year. The bean hornworm is an explosive pest in soybean production. The larvae eat the leaves too much, which can generally cause a 5-10% yield reduction. In severe cases, the bean sprouts are eaten into polished rods and the yield is reduced by more than 50%. For example, in 1990, the bean hornworm pest broke out in Xinxiang City, Henan Province, and the occurrence ...

Claims

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Application Information

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IPC IPC(8): C12N7/00A01N63/40A01P7/04C12R1/93
CPCC12N7/00A01N63/40C12N2710/14021C12N2710/14051C12N2710/14031Y02A50/30
Inventor 占智高关丽梅况文东
Owner INST OF MICROBIOLOGY JIANGXI ACADEMY OF SCI
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