Disease-prevention and growth-promoting Trichoderma harzianum and application thereof
A technology of bacteria strains and microorganisms, applied in the field of microbial pesticides, can solve the problems of drug resistance of pathogenic bacteria, sharp decline of biodiversity, and environmental pollution of chemical agents
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Embodiment 1
[0080] Embodiment 1, the isolation and identification of Trichoderma afroharzianum (Trichoderma afroharzianum) TM2-4 in Africa
[0081] 1.1 Strain isolation
[0082] The strain was isolated from soil samples of special habitat in Wulingyuan virgin forest in Hunan, China. Bacterial strains were separated by the dilution plate method, and the specific operation method was as follows: Weigh 10 g of the soil sample and put it into 90 ml of sterile water to vibrate evenly to obtain 10 -1 Concentration of bacterial solution, use a 1ml sterile pipette to draw 1ml 10 -1 Concentration of bacterial solution in a tube of 9ml sterile water, shake well that is 10 -2 Concentration of bacterial solution, the same method, sequentially diluted to 10 -4 . the above 10 -2 、10 -3 and 10 -4 The soil bacteria solution was spread on the PDA plate respectively, dried and placed upside down in an incubator at 25°C for 3-5 days. Pick a single colony that resembles Trichoderma and transfer it to...
Embodiment 2
[0093] Embodiment 2, African Harz Trichoderma (Trichoderma afroharzianum) TM2-4 is to the inhibition or heavy parasitism of testing pathogenic fungus
[0094] Pick Trichoderma afroharzianum (Trichoderma afroharzianum) TM2-4 bacterial cake with a diameter of 7 mm grown on the plate and the bacterial cake of the pathogenic fungus to be tested, and inoculate them symmetrically on the PDA medium at a position 2 cm away from the center, and the control does not inoculate African Trichoderma afroharzianum (Trichoderma afroharzianum) TM2-4 bacteria cake, only inoculate the bacteria cake of the pathogenic fungus tested, each treatment was repeated three times, and cultured at 25°C for 4 days to observe the growth of the pathogenic fungus. The pathogenic fungi tested included: Botrytis cinerea, Monilinia fructicola, Colletotrichum capsici, Fusarium oxysporum f.sp.cucumerinum.
[0095] Under confrontation culture conditions, Trichoderma afroharzianum TM2-4 can inhibit and cover tomato g...
Embodiment 3
[0096] Embodiment 3, Trichoderma afroharzianum (Trichoderma afroharzianum) TM2-4 in vitro control effect on tomato gray mold
[0097] 3.1 Preparation of Trichoderma afroharzianum TM2-4 sterile fermentation filtrate
[0098] Pick the Trichoderma afroharzianum TM2-4 grown on the plate with the inoculation loop and inoculate it into a 300mL Erlenmeyer flask containing 100mL of liquid fermentation medium, and cultivate it with shaking at 28°C and 180r / min for 2 days to obtain the seed liquid. % inoculum size Transfer the seed liquid to a 500mL Erlenmeyer flask containing 100mL liquid fermentation medium, 28°C, 180r / min shaking fermentation culture for 7d, and collect the fermentation broth. The fermented liquid was centrifuged at 12000 rpm for 10 min to take the supernatant, which was sterilized by filtration with a 0.22 μm microporous membrane to obtain a sterile fermented filtrate of Trichoderma afroharzianum TM2-4. The aseptic fermentation filtrate of Trichoderma afroharzianum...
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