Production of insect-proof sick wooden mold and its use

A technology of Trichoderma and Trichoderma harzianum, which is applied in the field of preparation of insect-resistant and disease-resistant Trichoderma, can solve the problems of difficult action of pests and short validity period, and achieve the effects of fast growth, thick hyphae, and shortened time course

Inactive Publication Date: 2006-05-31
杨谦
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AI Technical Summary

Problems solved by technology

Bacillus thuringiensis is the most productive and widely used microbial insecticide in the world, but its short-t

Method used

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Experimental program
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Effect test

Embodiment 1

[0034] The preparation method of insect-resistant and disease-resistant Trichoderma comprises: obtaining conventionally preserved Trichoderma harizianum, Agrobacterium tumefaciens AGL-1 and plasmid pPBH:

[0035] E. prepare conidia suspension by described Trichoderma harzianum;

[0036] f. transforming and activating the obtained Agrobacterium and] pPBH plasmid;

[0037] g. co-cultivate and transform the activated bacterium liquid of the above-mentioned conidia suspension, Agrobacterium and] pPBH plasmid;

[0038] h. Screen and purify transformants.

Embodiment 2

[0040] In the preparation method of the above-mentioned insect-resistant and disease-resistant Trichoderma, the preparation of the conidia suspension by Trichoderma harzianum is to wash the spores of Trichoderma harzianum with 5ml sterilized distilled water from the PDA plate of cultivating 5-7 days. Spores were counted with a hemocytometer, and then diluted with MM liquid medium to a spore concentration of 10 5 --10 6 pieces / ml.

Embodiment 3

[0042] In the preparation method of the above-mentioned insect-resistant and disease-resistant Trichoderma, the transformation and activation culture of the obtained Agrobacterium is to directly transform the pPBH plasmid into the Agrobacterium AGL-1 using a conventional freeze-thaw method, Pick a single Agrobacterium colony from the LB plate containing 50 μg / ml kanamycin and 50 μg / ml streptomycin, and inoculate it in 7 ml of 50 μg / ml kanamycin and 50 μg / ml streptomycin On LB liquid medium, cultivate overnight at 250rpm, 29°C, and dilute to OD with MM liquid medium containing 200μM vinyl syringone (AS) the next day 660 0.15, according to temperature and environment, continue to culture for 3-5 hours to OD 660 It is 0.6-0.8.

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Abstract

Production of anti-insect and disease-proof mold and its use are disclosed. The process includes taking hamycin, Agrobacterium tumefaciens AGL-1 and gene plasmid pPBH with thuricin bacillosporin stored in normal way, preparing conidiospore suspension, converting the obtained tumefaciens and pPBH plasmid, activation culturing, culturing the activated fungus liquid of conidiospore suspension, tumefaciens and pPBH plasmid mutually, converting, screening and purifying. It can be used to prepare anti-insect and disease-proof mold and insect prevention of plant.

Description

Technical field: [0001] The invention relates to a preparation method of an insect-resistant and disease-resistant Trichoderma, and an application of the Trichoderma to plant insect-resistant and disease-resistant. Background technique: [0002] Microorganisms are playing an increasingly important role in the field of biological control of plant diseases. Abroad, the United States has successfully transferred the Bt toxin gene to Pseudomonas bacteria (Pf), creating genetically engineered bacteria; Australian scientists have used biotechnology to , Agrobacterium actinomyces (K84) was improved into K1026 strain with persistent efficacy. In China, the genetic transformation system of bacteria such as Bacillus thuringiensis (Bt), Pseudomonas fluorescens (Pf), and Bacillus subtilis (Bs) has been established by using modern genetic engineering technology, and a set of strain isolation and screening suitable for my country's national conditions has been created. , gene identificati...

Claims

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Application Information

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IPC IPC(8): C12N1/00A01N63/02C12N1/14C12N1/20
Inventor 杨谦
Owner 杨谦
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