Method for protection of pleurotus eryngii strain by using uracil auxotrophy
A technology of auxotrophy and uracil is applied in the field of protection of Pleurotus eryngii strains using uracil auxotrophy, which can solve the problems of being easily counterfeited and achieve the effect of low cost and simple method
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Embodiment 1
[0032] Acquisition of Uracil Auxotrophic Monospores of Pleurotus eryngii
[0033] The auxotrophic strain NX2-0 of Pleurotus eryngii was crossed with monokaryotic strain DX8 of Pleurotus eryngii to obtain dikaryotic strain SXN1 for fruiting. After fruiting, select two single fruiting bodies with complete mushroom shape, large size, thick flesh, and no pests and diseases, and collect naturally ejected spores in a sterile plate in an ultra-clean bench. Soak the collected spores in an EP tube filled with sterile water, and then use sterile water to gradiently dilute until there are 2-3 single spores in each field of view during microscopic examination. The spore liquid was spread on the PDA medium and cultured in a biochemical incubator at 25°C. Generally, germinated spores can be seen within 1 week. Transfer the germinated spores to the PDA medium, and after 10 days of cultivation, put them on a microscope for inspection. The hyphae without lock-like associations are those that...
Embodiment 2
[0035] Preparation of Pleurotus eryngii Uracil Auxotrophic Dikaryon Strain
[0036] The monokaryotic hyphae obtained from the germination of the uracil auxotrophic monospore strain were hybridized on the MMU medium. Cultivate for about two weeks, see that the parental hyphae have fused, pick out the mycelium at the fusion point for microscopic examination, and the mycelia with lock-like joints are uracil auxotrophic dikaryotic mycelia (UX1-5 (CGMCC NO.13693, the preservation date is 2017 On March 13, 2009, the preservation unit CGMCC (General Microorganism Center of China Microbiological Culture Collection Management Committee, No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing), suggested that the classification be named: Pleurotus eryngii (Pleurotus eryngii)) as 1 -6 is obtained by crossing 1-43, and 1-6 and 1-43 are obtained by collecting single spores from fruiting bodies obtained by crossing NX2-0 and DX8).
Embodiment 3
[0038] Validation of 5'-fluoroorotic acid in uracil auxotrophic dikaryotic strain
[0039] The obtained Pleurotus eryngii uracil auxotrophic dikaryotic strains were inoculated on different mediums MM, MMU, and MMUF, and these homozygotes could grow on MMU medium and MMUF medium but could not grow on MM medium. Such as image 3As shown, the uracil auxotrophic dikaryotic strain UX1-5 could grow on MMU and MMUF but not on MM.
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