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Novel method for producing steinernema carpocapsae on large scale

The technology of a new method of St. chinensis and a new method can be applied in the field of biological control and can solve the problems of affecting the quality of nematodes, limiting the large-scale cultivation of entomopathogenic nematodes and the like.

Inactive Publication Date: 2017-08-04
HUNAN UNIV OF TECH
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Problems solved by technology

The above and other reasons seriously limit the large-scale cultivation of entomopathogenic nematodes and affect the quality of nematodes

Method used

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  • Novel method for producing steinernema carpocapsae on large scale

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Experimental program
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Embodiment 1

[0007] Embodiment 1 selects 1000 healthy last-instar larvae of S. mellonella, puts the sponge with the small tortrix nematode into deionized water, stirs for several times, picks out the sponge, and detects with a microscope that the density of nematodes is 1,000,000 tails / ml. Inoculate the surface of 1000 wax moths. One day later, when the wax moth dies, select the dead wax moth and culture it for 10 days under a constant temperature of 25 degrees Celsius and a humidity of 90% in a constant humidity environment, and the nematodes obtained are the F0 generation. The nematodes of the F0 generation were extracted from the dead body of Mellonella mellonella with sterile deionized water, and then the nematodes were washed with sterile physiological saline for 3-5 times, and then centrifuged. Inoculate 500,000 nematodes of the F0 generation with no miscellaneous bacteria into a 15mm nutrient agar culture dish that has been cultured for 2 days with Bacillus nematophila. The culture ...

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Abstract

The invention provides a novel method for stably producing a steinernema carpocapsae F3 generation based on the principle of mutualistic symbiosis of xenorhabdus and nematode and through combination of non-in-vitro culture and in-vitro culture. Healthy galleria mellonella final instar larvae are selected and inoculated with steinernema carpocapsae, and the obtained nematode is the Fo generation; a nutrient agar culture dish with xenorhabdus cultured for two days is inoculated with the nematode Fo generation, and the F1 generation is obtained; finally, an amplification sponge culture medium is inoculated with the F1 generation, culture is carried out for 14 days, and a large quantity of F3 generation is obtained to serve as commercial nematode. The commercial nematode produced through the method is the F3-generation nematode, the production period is short, and the quality is stable.

Description

technical field [0001] The invention belongs to the biological control technology, and in particular relates to a new method for stably producing the F3 generation of S. toricutellae by utilizing the principle of symbiosis between Bacillus nematophila and nematodes, and combining non-isolated culture and isolated culture. Background technique [0002] Steineria minor is an entomopathogenic nematode that takes insects as hosts. The entomopathogenic nematodes in the invasion stage can find hosts independently, and the nematodes that invade the host release entomopathogenic bacilli from the intestinal tract, and the bacilli multiply in the host in large numbers, resulting in the rapid death of the host. The entomopathogenic nematodes commonly used to control pests in the world mainly belong to the genus Steineria and Heterobradida. There are 10 species of Steiner's nematodes and 3 species of Heterobacterium nematodes. In the biopesticide market in industrialized countries, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/033
CPCA01K67/033
Inventor 李小龙张凤琴何农跃
Owner HUNAN UNIV OF TECH
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