Beauveria bassiana strain with pathogenicity on alissonotum impressicolle arrow and application thereof
A technology of pathogenicity and Beauveria bassiana is applied in the field of biotechnology control to achieve the effects of rapid growth, high spore germination rate, and large spore production
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Embodiment 1
[0021] Isolation and identification of pathogenic bacteria.
[0022] Submerge the collected naturally infected corn borer worms in 75% alcohol for 3-5 seconds, then sterilize the surface with 0.1% mercury chloride for 3 minutes, rinse with sterilized water for 3 times, and under aseptic conditions, use a sterilized knife to kill the infected Cut worms into 3mm square pieces and inoculate them on a PDA plate, put them in an incubator at 28°C and cultivate them. After the mycelium grows, perform single-spore isolation and culture according to conventional methods to obtain pure Beauveria bassiana strain Bb-YMM, and transfer Cultivate on PDA slant for 7 days and store in refrigerator at 4°C.
[0023] Morphological identification:
[0024] Cultivate the isolated and purified Beauveria bassiana strain on a flat PDA medium with a diameter of 90mm at 27±1°C, pick white hyphae after 4-5 days, and observe the hyphae and distribution of the strain under a microscope. Morphology of spo...
Embodiment 2
[0029] Biological characteristics of Beauveria bassiana Bb-YMM.
[0030] Determination of colony growth rate and spore production: inoculate Beauveria bassiana strain Bb-YMM on the center of PDA plate medium by drop method, culture in an incubator at 27±1°C, and use cross method The colony diameter was measured once a day, and three replicates were set for each treatment, and the total observation time was 15 days. At the same time, record the start time of spore production, use a puncher with a diameter of 1.5cm, and take a small plate from the center of the colony to the edge 1 / 2, take out the plate and place it in a container containing 20mL of 0.1% Tween-80 sterile water. In the Erlenmeyer flask, oscillate to fully disperse the spores and dilute them appropriately, measure the spore concentration with a hemocytometer and convert it into the sporulation per unit area.
[0031] Determination of spore germination rate: culture the strain for 10 days, collect conidia with ste...
Embodiment 3
[0040] The pathogenicity of Beauveria bassiana (Beauveria bassiana) Bb-YMM to beetle larvae was determined by dipping method.
[0041] Scrape a certain amount of Beauveria bassiana (Beauveria bassiana) Bb-YMM spores from the PDA plate and place them in sterilized 0.1% Tween-80 solution to prepare 10 7 Spores / mL of spore suspension; impregnate the healthy and active 3rd instar larvae of the cane beetle, wherein the larvae of the beetle are shown in figure 2 As shown, at the same time gently flip the worm body, take it out after 10s and put it in a sterilized can bottle (inner diameter 6cm, height 11cm), put a small section of sugarcane stem for it to eat, and then fill the bottle with soil (soil passed through the oven High temperature sterilization, humidity about 20%). For each treatment, 20 larvae were reared and repeated 4 times, and treated with 0.1% Tween-80 sterile aqueous solution as a control. After treatment, place it in a dark 26±1°C constant temperature incubator...
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