A kind of black fungus for preventing and treating fungal diseases and application thereof
A black fungus and strain technology, applied in the application, fungi, pest control and other directions, to achieve the effect of good growth promotion, low cost, inhibition of occurrence and spread
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Embodiment 1
[0055] Isolation and Identification of DK6 Strain
[0056] 1. Isolation location, isolation method (morphological identification of traditional microbiology)
[0057] Microbial characteristics
[0058] Isolation of DK6 strain: The applicant isolated and screened a basidiomycete strain with strong control effect on Botrytis cinerea from corn leaves in Shiyan City, Hubei Province, China, and the applicant named it DK6. The separation and screening of the DK6 bacterial strain of the present invention adopt methods such as tissue separation method, dilution plate method, plate antagonism, in vitro test, strain identification work with reference to " fungus identification handbook " (Wei Jingchao, 1979), " morphology and classification of fungi " (Dai Fanglan, 1987) and other monographs in the test method. The DK6 strain was cultured at 25°C for 10 days, the genomic DNA was extracted by the CTAB method (Taylor et al, 1993), and the rDNA-ITS region was amplified using the general ...
Embodiment 2
[0070] DK6 strain and Botrytis cinerea plate confrontation test:
[0071] Use a hole punch with an inner diameter of 9mm to punch the activated Botrytis cinerea and connect it to one side of the PDA plate, and the other side to connect the DK6 strain (with an inner diameter of 9mm). Botrytis cinerea was inoculated and the other side was not inoculated as a control, and repeated 3 times. After 3 to 4 days, observe the confrontation result. See the test results Figure 4 .
[0072] The results of the confrontation test show that the DK6 strain can cover Botrytis cinerea and continue to grow, and it grows more densely on Botrytis cinerea. It is preliminarily estimated that the DK6 strain can compete with Botrytis cinerea for nutrients or can continue to grow by using Botrytis cinerea itself or its metabolites. The mode of action is nutrient competition, and it is a strain with strong inhibitory effect on Botrytis cinerea.
Embodiment 3
[0074] Preliminary Analysis of the Antagonistic Mechanism of Strain DK6
[0075] (1) Effect of DK6 fermentation product on the growth rate of Botrytis cinerea
[0076] Use a hole punch with an inner diameter of 9mm to take out the DK6 bacterial block, transfer it to 100mL PDB, and shake the bacteria for 5 days at 25°C and 160rpm. The prepared fermentation liquid was centrifuged at 8,000 r / min for 10 min, and the supernatant was suction-filtered with a 0.22 μm microporous membrane to obtain a sterile fermentation liquid, namely the fermentation supernatant. Fermentation supernatant was added in PDA by 1% (volume ratio), 5%, 10%, poured plate, each plate was connected with gray mold mycelia pieces with a diameter of 9mm, each treatment was repeated 3 times, 3 parallel experiments , cultivated at 20°C, measured the colony diameter of Botrytis cinerea on 3d, 5d, and 7d and recorded relevant data. The test result is Figure 5 , Figure 6 .
[0077] Figure 5 The results showe...
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