Method for breeding selection of insecticidal fungus with relatvely stable pathogenicity

A technology of pathogenicity and entomogenic fungi, which is applied in the field of biological control to achieve the effect of high pathogenicity and stable pathogenicity

Inactive Publication Date: 2007-10-10
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a method that is simpler than the protoplast fusion method and does not require physical and chemical mutagenesis of the parental strain has not been reported

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] (1) 32 monospore strains are isolated from 8 Paecilomyces cicadae (Miq.) Samson] bacterial strains collected from the whole country, and the monospores of each entomogenous fungus that are isolated are cultivated in chlorate potatoes Induce mutations on the base (KPS);

[0026] (2) From the chlorate potato medium (KPS), the chlorate-resistant horn was isolated by punching, and identified on the basal medium (MM) and the chlorate potato medium (KPS);

[0027] (3) Any angular change that grows well on the chlorate potato medium (KPS) and has only weak hyphae on the basal medium (MM) at the same time is a true nitrate utilization-deficient (nit) mutant;

[0028] (4) The selected nit strains were pair-cultured on basal medium (MM) with an interval of 3-5 cm at 25°C for about 21 days in dark to observe the affinity between the strains. If it is compatible, a vigorous mating cluster will be formed between the bacterial colonies; if it is not compatible, there will be no spec...

Embodiment 2

[0050] (1) Pour into sterile water containing 0.05% Toween-80, on the plate colony of Paecilomyces farinosus (Holm.ex Gray) Brown & Smith] that has formed conidia, and gently press it with a glass scraper Spores were washed off the colony surface. Transfer the spore suspension into a small Erlenmeyer flask with several glass beads, shake it well to disperse it, and then filter it with sterile glass wool to remove mycelium. Dilute the spore suspension to about 300 spores per ml, suck 0.1ml of this solution onto a base medium (MM) plate with a diameter of 9cm, spread it evenly with a glass scraper, incubate at 25°C, observe under a microscope to confirm and mark the concentricity Radiation-grown monospore colonies, which are visible to the naked eye, are isolated and cultured as monospore strains;

[0051] (2) isolate and obtain 26 monospore strains from the collected Paecilomyces farinosus (Holm.ex Gray) Brown & Smith] bacterial strain, each single spore that separates is obta...

Embodiment 3

[0075] (1) Pour sterile water containing 0.05% Toween-80 on the colony of Paecilomyces fumosoroseus (Wize) Brown & Smith] that has formed conidia, and gently press the surface of the colony with a glass scraper to wash it off spore. Dry the spore suspension into a small Erlenmeyer flask with several glass beads, shake it fully to disperse it, and then filter it with sterile glass wool to remove mycelium. Dilute the spore suspension to about 300 spores per ml, suck 0.1ml of this solution onto a base medium (MM) plate with a diameter of 9cm, spread it evenly with a glass scraper, incubate at 25°C, observe under a microscope to confirm and mark the concentricity Radiation-grown monospore colonies, which are visible to the naked eye, are isolated and cultured as monospore strains;

[0076](2) isolate and obtain 20 single-spore bacterial strains from collected Paecilomyces fumosoroseus (Wize) Brown & Smith] bacterial strains, and induce each single-spore strain obtained by separat...

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Abstract

This invention discloses a method for breeding pesticidal fungi with stable pathogenicity. The method comprises: (1) separating monospores of pesticidal fungi, mutagenizing on chlorate potato medium (KPS), punching to separate anti-chlorate mutants, and identifying with basic medium and KPS medium to obtain nitrate-utilization auxotrophic mutants (nit); (2) selecting nit mutants with high vegetative compatibility to obtain parental strains, collecting zygospores, and culturing to obtain heterocaryon; (3) staining the heterocaryon with Hoechest 33258, and observing to identify heterocaryosis. The pesticidal fungi have stable pathogenicity, and can prevent and treat pests in agriculture, forestry and stockbreeding.

Description

technical field [0001] The invention relates to a method for breeding insect-growing fungi in the field of biological control. Background technique [0002] The use of fungal natural enemies of insects to develop fungal preparations to control insect pests is an important content of biological pest control. However, it always faces two difficult problems, one of which is the instability of fungal strains' pathogenicity to pests. Generally, the pathogenicity of a production strain will decrease after three consecutive generations. This is because the nuclei of different genotypes are easy to separate during the quasi-sexual recombination process, resulting in the so-called "strain degeneration", which brings a lot of trouble to production and application. [0003] Theoretically speaking, physical and chemical mutagenesis, heterokaryon synthesis and quasi-recombination are the three basic ways for most entomopathogenic fungi of the subphylum Agnostica to cultivate highly viru...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01N53/04A01P5/00A01P7/00
Inventor 刘爱英赵杰宏邹晓赵德刚韩洁文庭池
Owner GUIZHOU UNIV
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