A Java Cordyceps strain bd01 and its application
A technology of Cordyceps and strains, which is applied in the field of agricultural microorganisms, can solve the problem of fewer microorganisms, and achieve the effect of reducing the impact and benefiting the yield and quality
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Embodiment 1
[0032] Isolation, Purification and Identification of Cordyceps Javanese Bd01 Strain
[0033] 1. Isolation and purification of bacterial strains
[0034] It was isolated from the dead beetle larvae of the coffee beetle larvae, which died naturally. microbial pesticides. Adopt the following steps to carry out separation and purification:
[0035] 1. First, rinse the larvae of the beetle longicorn beetle that died of the disease with 75% alcohol, then rinse 3 times with sterile water, and finally dry the water with sterilized filter paper for later use.
[0036] 2. Sterilization: All experimental equipment such as petri dishes, centrifuge tubes, test tubes, and pipette tips and sterile water are sterilized by high pressure at 0.1MPa, 121°C, and sterilized for 30 minutes.
[0037] 5. Preparation of culture medium
[0038] Medium for separation and purification: PDA medium, including: 200g of peeled potatoes, cut into pieces and boiled with water for 20min, filtered through dou...
Embodiment 2
[0067] The influence of embodiment 2 different environmental conditions on bacterial strain vegetative growth
[0068] The Cordyceps javanica strain was inoculated on PDA medium and cultured for 2 weeks for the optimization of the following conditions.
[0069] 1. Effects of different media on vegetative growth and sporulation of Cordyceps javanica strain
[0070] The strains were cultured in five mediums of PDA, PPDA, SDAY, SMAY, and Czapek, and the colony diameter was measured every 2 days for a total of 10 days. At the same time, the initial sporulation time recorded on different media was observed, and the sporulation was measured after 14 days. quantity. Use a hole puncher with a diameter of 5 mm to take 3 colonies (at the same position as the center of the petri dish) and put them in 2 mL of sterile water containing 0.1% Tween-80, shake them fully, measure the number of spores with a hemocytometer, and calculate the amount of sporulation. Select the most suitable mediu...
Embodiment 3
[0088] The influence of embodiment 3 different environmental conditions on bacterial strain pathogenicity
[0089] Preparation of spore suspension: according to the screening conditions in Example 2, the Cordyceps javanica strain Bd01 was inoculated in PPDA medium, and cultured in a constant temperature incubator at 26°C for 2 weeks;
[0090] The Cordyceps javanica strain Bd01 that has been cultured for 2 weeks and completely sporulated was washed out with sterile water containing 0.1% Tween-80, stirred evenly with a magnetic stirrer, counted by a hemocytometer, and prepared into a spore suspension of the required concentration.
[0091] 1. Pathogenicity of strains at different temperatures
[0092] Select 20 healthy long-tailed beetle larvae with the same shape and size, and soak them at a concentration of 1×10 8 After spores / mL of spore suspension for 30 s, they were inserted into finger tubes (1.2×5 cm), and each treatment was repeated 3 times. Put them into the incubator...
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