A preparing method of a hyperparasitic fungal inoculant for sclerotinia sclerotiorum
A technology of fungal agent and sclerotinia, applied in botany equipment and methods, biocides, fungicides, etc., can solve the problems of food safety impact, unsatisfactory control effect, high cost, etc.
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Embodiment 1
[0023] Isolation culture:
[0024] Collect rhizosphere soil samples, add sclerotia to 30 grains / 100 g soil samples and mix well, trap parasites at a constant temperature of 25°C; take them out after 3 weeks, rinse with tap water, sterilize the surface with 1% NaCIO for 1 minute, and fully sterilize with sterile water. After rinsing, put it on a filter paper sheet placed in a petri dish, and after 2 weeks of moisturizing culture at 25°C, check the hyperparasites on the surface of the sclerotium under a stereoscope, pick out monospores and culture them on a PDA plate, and purify and separate them.
[0025] Liquid fermentation:
[0026] Test tube culture: use solid potato grape agar (PDA) medium, inoculate the hyperparasitic fungal strain on the test tube medium, and culture at 26°C for 4 days;
[0027] Expansion culture: use liquid potato dextrose (PD) medium to inoculate the hyperparasitic fungal strains in the test tubes into Erlenmeyer flasks, place them on a shaker at 26°C ...
Embodiment 2
[0032] The process steps are as in Example 1, the rhizosphere soil sample is collected in the separation and cultivation step, sclerotia is added in 40 grains / 100 g of the soil sample and mixed evenly, parasitic bacteria are trapped at a constant temperature of 25°C, and taken out after 3.5 weeks. In the liquid fermentation step, the fermentation culture uses glucose and urea as carbon and nitrogen sources respectively, and the C / N ratio is 12:1. In the liquid fermentation step, the culture in the Erlenmeyer flask is added to the fermentation broth at a ratio of 7%. In the preparation step of the bacterial agent, the bacterial slurry obtained by liquid fermentation is mixed with diatomaceous earth at a ratio of 1.2:1 (V / W). The effective amount of live bacteria is 4-5 billion / g.
Embodiment 3
[0034] The process steps are as in Example 1, the rhizosphere soil sample is collected in the separation and cultivation step, 50 grains / 100 grams of soil sample are added to the sclerotia and mixed evenly, the parasitic bacteria are trapped at a constant temperature of 25°C, and taken out after 4 weeks. In the liquid fermentation step, the fermentation culture uses glucose and urea as carbon and nitrogen sources respectively, and the C / N ratio is 15:1. In the liquid fermentation step, the culture in the Erlenmeyer flask was added to the fermentation broth at a ratio of 8%. In the preparation step of the microbial agent, the bacterial slurry obtained from liquid fermentation is mixed with diatomaceous earth at a ratio of 1.5:1 (V / W). The effective amount of live bacteria is 3-4 billion / gram.
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