Metarhizium rileyi CDTLJ1 and application thereof
A technology of Metarhizium anisopliae and Layleigh, which is applied in the field of biotechnology control, can solve the problems of the short invasion time of Spodoptera frugiperda in my country and the limited research on biological control.
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Embodiment 1
[0027] Isolation and identification of pathogenic bacteria.
[0028] Naturally infected larvae of Spodoptera frugiperda were collected from the Lijian Research Base of Guangxi Academy of Agricultural Sciences, Wuming District, Nanning City, Guangxi Zhuang Autonomous Region. Under sterile conditions, a small amount of spores were picked up with an inoculation needle and inoculated on Sabouraud's by streaking method. Maltose agar yeast medium (SMAY) (maltose 40g, peptone 10g, yeast extract 10g, agar 18g, distilled water 1000mL), at temperature (26±1)℃, photoperiod L:D=12h:12h, relative humidity (80 ±5)% in a constant temperature incubator. After 7 days of culture, pick a small amount of spores to a new medium for purification and culture for 15 days, place the isolated and purified conidia of the strain in 20% glycerol, and store them in a -80°C refrigerator for later use.
[0029] Morphological identification:
[0030] The isolated and purified strains were cultured on a flat...
Embodiment 2
[0038] Biological characteristics of Metarhizium rileyi CDTLJ1.
[0039] Determination of bacterial colony growth rate and spore production: Inoculate Metarhizium rileyi CDTLJ1 on the center of PDA, SDAY, PPDA, SMAY, and Czapek plate culture medium by spot method, and place it at a temperature of (26±1)°C. Photoperiod L:D=12h:12h, cultivate in a constant temperature incubator with relative humidity (80±5%), measure the diameter of the colony every day with the cross method, set 3 repetitions for each treatment, and observe for 15 days in total. Simultaneously record the start time of spore production. From the start of spore production, take out the whole bacterial colony plate every day and place it in an Erlenmeyer flask containing 20mL of sterilized 0.05% Tween-80 solution, shake to make the spores fully disperse, measure the spore concentration with a hemocytometer Converted into sporulation per unit area.
[0040] The mensuration of spore germination rate: bacterial stra...
Embodiment 3
[0056] The pathogenicity of different spore concentrations of Metarhizium rileyi (Metarhizium rileyi) CDTLJ1 to the third instar larvae of Spodoptera frugiperda was determined by soaking insects.
[0057] A certain amount of Metarhizium rileyi CDTLJ1 spores were scraped from the SMAY plate and placed in sterilized 0.05% Tween-80 solution to prepare 5 concentration gradient spore suspensions (1×10 4 , 1×10 5 , 1×10 6 , 1×10 7 , 1×10 8 spores / mL); select healthy and lively 3rd instar larvae of Spodoptera frugiperda with the same size, put them into the prepared spore suspension, take them out after soaking for 5s, put them on sterile filter paper to absorb excess water on the surface of the worms, and place It is placed in a sterilized petri dish (d=7.5 cm) with wet filter paper on the bottom of the dish, and fresh corn leaves are placed in the dish for feeding. Each treatment of 20 larvae, single rearing, repeated 3 times, treated with 0.05% Tween-80 sterile aqueous solutio...
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