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A method for protecting Flammulina velutipes strains by using uracil auxotroph

An auxotrophic and uracil technology, which is applied in the preparation of fungi and hybrid cells, can solve problems such as being easily counterfeited, and achieve the effects of low cost and simple method.

Active Publication Date: 2020-08-04
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the breeding of strains can only be achieved by relying on long-term accumulation, which involves a lot of manpower, material resources and time costs.
However, because Flammulina velutipes can be reproduced asexually, the production process of its strains is reproducible, making it easy to be counterfeited

Method used

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  • A method for protecting Flammulina velutipes strains by using uracil auxotroph
  • A method for protecting Flammulina velutipes strains by using uracil auxotroph
  • A method for protecting Flammulina velutipes strains by using uracil auxotroph

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Experimental program
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Effect test

Embodiment 1

[0033] Acquisition of Uracil Auxotrophic Monospores of Flammulina velutipes

[0034] The auxotrophic strain NG1-92 of Flammulina velutipes was crossed with monokaryotic strain DG1-29 of Flammulina velutipes to obtain dikaryotic strain SGN2 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 a...

Embodiment 2

[0036] Preparation of uracil auxotrophic dikaryotic strain of Flammulina velutipes

[0037] 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 mycelium of the parents has fused, pick out the mycelium at the fusion point for microscopic examination, and the mycelia with lock-like joints are uracil auxotrophic dikaryotic mycelia (VG1-2 (CGMCC NO.12877, the preservation date is 2016 On September 19, 2009, the depository unit, China Microbiological Culture Collection Management Committee General Microbiology Center, the preservation address was No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, and the proposed classification was named: Flammulina velutipes) for 2-6 and 2-33 is obtained by crossing, and 2-6 and 2-33 are obtained by collecting single spores from fruit bodies obtained by crossing DG1-29 and NG1-92).

Embodiment 3

[0039] Validation of 5'-fluoroorotic acid in uracil auxotrophic dikaryotic strain

[0040] The obtained uracil auxotrophic dikaryotic strains of Flammulina velutipes were inoculated on different mediums MM, MMU, and MMUF, and these homozygous strains could grow on MMU medium and MMUF medium but could not grow on MM medium. Such as image 3 As shown, the uracil auxotrophic dikaryotic strain VG1-2 could grow on MMU and MMUF but not on MM.

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Abstract

The invention relates to a method for protecting the flammulina velutipes strain by utilizing uracil auxotroph. The method comprises the following steps: hybridizing uracil auxotroph monokaryon NG1-92 of flammulina velutipes with a monokaryon wild type strain of flammulina velutipes, and after fruiting, carrying out single spore isolation, thus obtaining a monokaryon strain; inoculating the monokaryon strain to 5-fluoroorotic acid screening culture medium for culturing, carrying out identification, thus obtaining the uracil auxotroph monokaryon strain, and carrying out mon-mon crossing to obtain a uracil auxotroph dikaryon strain. The method provided by the invention is simple and is low in cost, the strain obtained by adopting the conventional strain separation method can not grow and propagate normally, and thus the flammulina velutipes strain is effectively protected.

Description

technical field [0001] The invention belongs to the field of protection of Flammulina velutipes strains, in particular to a method for protecting Flammulina velutipes strains by using uracil auxotrophs. Background technique [0002] Flammulina velutipes is one of the widely cultivated edible fungi in my country and has important food and medicinal value. The development of cultivation and production of Flammulina velutipes must depend on bacterial classification to a large extent. However, the breeding of strains can only be gained by relying on long-term accumulation, which involves a lot of manpower, material resources and time costs. However, because Flammulina velutipes can be asexually reproduced, its strain production process is reproducible, making it easy to be counterfeited. [0003] Auxotrophic strains have lost the ability to synthesize certain substances due to gene mutations, and cannot grow normally in the basic medium, and must be supplemented with certain s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/04C12N1/14
CPCC12N1/14C12N15/04
Inventor 鲍大鹏茅文俊卢绪志张光忠吴莹莹李燕唐利华高英女杨瑞恒龚明
Owner SHANGHAI ACAD OF AGRI SCI
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