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Conidiobolus cytoplasm soluble extract with insecticidal killing activity and use thereof

A technology of insecticidal activity and cytoplasm of ear mold, which is applied in the field of soluble extract of ear mold cytoplasm, can solve the problems of limited practical application, difficulty in large-scale production, and limited number of conidia

Active Publication Date: 2019-05-03
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Entomogenous entomopathogenic fungi are different from hyphosporium fungi such as Beauveria bassiana. The number of conidia produced is limited, and it is difficult to carry out large-scale production, which limits the practical application of this kind of valuable fungal resources

Method used

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  • Conidiobolus cytoplasm soluble extract with insecticidal killing activity and use thereof
  • Conidiobolus cytoplasm soluble extract with insecticidal killing activity and use thereof
  • Conidiobolus cytoplasm soluble extract with insecticidal killing activity and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Otomycetes activated on enriched (v / v: 9% pure milk) Sabouraud medium (w / v: 3% glucose, 1.2% yeast powder, 1.2% peptone), cultivated in the dark at 23°C for 4 days , Pick about 100mg of fresh bacteria blocks (2-3mm×2-3mm several pieces) and add them to 100mL Sabouraud’s liquid medium (w / v, 2% glucose, 0.9% yeast powder, 0.9% peptone), at 24°C, Shake the bacteria at 190rpm for 2 days, passage 1 time (interval 2 days), fermented bacteria liquid with a mycelium concentration of about 16mg / mL (dry weight w / v), filter the liquid with filter paper, take 100mg of mycelia and Grind and crush the cells with a pulper, add 1 mL of phosphate buffer (pH7.7), vortex the mixture for 0.8 minutes, repeat three times, centrifuge at 12000g for 18 minutes at 5°C, and obtain the soluble protein extract of Atomyces cytoplasm in the supernatant , the protein concentration in the cytoplasmic extract was about 984 μg / ml.

Embodiment 2

[0026] Otomycetes activated on enriched (v / v: 10% pure milk) Sabouraud's medium (w / v: 4% glucose, 1% yeast powder, 1% peptone), cultivated in the dark at 24°C for 5 days , Pick about 100mg of fresh bacterial blocks (2-3mm×2-3mm several pieces) and add them to 100mL Sabouraud’s liquid medium (w / v, 4% glucose, 1% yeast powder, 1% peptone), at 24°C, Shake the bacteria at 200rpm for 3 days, subculture 1 time (interval 3 days), ferment the bacteria liquid with a mycelium concentration of about 25mg / mL (dry weight w / v), filter the liquid with filter paper, take 100mg of mycelia and Grind the cells with a pulper, add 1mL phosphate buffer (pH7.8), vortex the mixture for 1 minute, repeat three times, centrifuge at 12000g for 20 minutes at 4°C, and obtain the soluble protein extract of Atomyces cytoplasm from the supernatant , the protein concentration in the cytoplasmic extract was about 1579 μg / ml.

Embodiment 3

[0028]Otomycetes activated on enriched (v / v: 12% pure milk) Sabouraud medium (w / v: 3% glucose, 0.9% yeast powder, 0.9% peptone), cultured in the dark at 25°C for 4 days , Pick about 100mg of fresh bacterial blocks (2-3mm×2-3mm several pieces) and add them to 100mL Sabouraud’s liquid medium (w / v, 8% glucose, 1.1% yeast powder, 1.1% peptone), at 20°C, Shake the bacteria at 210rpm for 2 days, subculture twice (transfer to Sabouraud medium every 2 days), and filter the fermentation broth with mycelium concentration of about 20 mg / mL (dry weight w / v) with filter paper After liquid, take 100 mg of mycelia and crush the cells with a homogenizer, add 1 mL of phosphate buffer (pH7.9), vortex the mixture for 1.2 minutes, repeat 4 times, centrifuge at 12000 g for 25 minutes at 3 °C, and the supernatant The cytoplasmic soluble protein extract of Atomyces auriticum was obtained, and the protein concentration in the cytoplasmic extract was about 1255 μg / ml.

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Abstract

The invention relates to a conidiobolus cytoplasm soluble extract with insecticidal killing activity and use thereof, which belong to the technical field of a pesticide. A preparation method comprisesthe followings steps: 1) conidiobolus activation; 2) liquid shake culture; 3) mycelial cell wall-breaking; and 4) soluble cytoplasmic protein extraction. The above-mentioned conidiobolus cytoplasm soluble extract has excellent insecticidal killing activity, and the mortality rate of thegreater wax moth larva and the pine wood nematode imago reaches 100% and 96.4% under the optimal conditions. Thepreparation is simple, and the soluble extract can be applied to the control of agricultural and forestry pests.

Description

technical field [0001] The invention belongs to the technical field of pesticides, and in particular relates to a soluble extract of auricularia cytoplasm with nematicide and insecticide activity and its application. Background technique [0002] Agricultural and forestry crops are eroded by different kinds of diseases and insect pests all year round, resulting in huge loss of yield. Among them, there are many kinds of nematodes, most of which are parasites, with more than 10,000 species, including pathogens of major agricultural and forestry crops and animals, such as root-knot nematodes that harm crops and pine wood nematodes that cause the death of large areas of pine forests. Forestry industry development poses a huge threat. At present, chemical pesticides are generally used for prevention and control, but due to the requirements of environmental protection, some nematicides are gradually eliminated, such as highly toxic carbofuran. Therefore, it is necessary to devel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P1/02A01P5/00A01P7/04C12R1/645
Inventor 周湘郭恺王江宏陈莎妮
Owner ZHEJIANG FORESTRY UNIVERSITY
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