Method for protecting lentinus edodes strain through uracil auxotroph

An auxotrophic, uracil-based technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, fungi, etc., can solve the problems of being easy to be counterfeited, and achieve the effect of low cost and simple method

Active Publication Date: 2016-07-27
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 shiitake mushrooms reproduce asexually, the production process of the mushrooms is reproducible, making it easy to be counterfeited

Method used

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  • Method for protecting lentinus edodes strain through uracil auxotroph
  • Method for protecting lentinus edodes strain through uracil auxotroph
  • Method for protecting lentinus edodes strain through uracil auxotroph

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Obtaining of auxotrophic monospores of Lentinus edodes uracil

[0035] The auxotrophic strain 423-9 of lentinus edodes was crossed with the monokaryon strain W661 of lentinus edodes to obtain the binuclear strain Z2 for fruiting. After fruiting, choose two single fruiting bodies with complete mushroom shape, large size, thick flesh and no pests and diseases, and collect the naturally ejected spores with a flat plate in a sterile bell jar in an ultra-clean platform. Soak the collected spores in an EP tube filled with sterile water, and then gradually dilute with sterile water until there are 2-3 single spores in each field of view during microscopic examination. Spread the spore liquid on the PDA medium and place it in a 25°C biochemical incubator for cultivation. Germination of spores can generally be seen within 2 weeks. Transfer the germinated spores to the PDA medium, after 10 days of cultivation, check on the microscope, the hyphae with no lock-like association are t...

Embodiment 2

[0037] Preparation of Lentinus edodes uracil auxotrophic binuclear strain

[0038] The monocytic hyphae obtained from the germination of the uracil auxotrophic monospore strain were hybridized on MMU medium. After culturing for about two weeks, the parent hyphae have been fused. Pick out the hyphae at the fusion site for microscopic examination. The uracil auxotrophic binuclear hyphae with lock-like associations are found. (UH9 is the hybridization of Z2-14 and Z2-97, UH21 is the hybridization of Z2-18 and Z2-97. Z2-14, Z2-18, Z2-97 are the fruiting bodies of the hybridization of W661 and 423-9 to collect monospores get).

[0039] The UH9 preservation number is: CGMCCNO.11916, and the classification designation: Lentinulaedodes is preserved on January 29, 2016. Preservation unit: China Common Microbial Species Collection Management Center CGMCC. Preservation address: No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing.

[0040] The preservation number of UH21 is: CGMCCNO....

Embodiment 3

[0042] Verification of uracil auxotrophic binuclear strain 5'-fluoroorotic acid

[0043] The obtained Lentinus edodes uracil auxotrophic binucleosis strains were inoculated into different media MM, MMU, MMUF, and these homozygotes could grow on MMU medium and MMUF medium but not on MM medium. Such as image 3 The indicated uracil auxotrophic binuclear strain UH21 can grow on MMU and MMUF but not on MM.

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Abstract

The invention relates to a method for protecting a lentinus edodes strain through uracil auxotroph.The method comprises the steps that a lentinus edodes uracil auxotroph monokaryon 423-9 and a lentinus edodes monokaryon wild-type strain are crossbred, and single spore isolation is carried out after fruiting to obtain a monokaryon strain; a 5-fluororotic acid screening medium is inoculated with the monokaryon strain for culture, authentication is carried out to obtain a uracil auxotroph monokaryon strain, and then a uracil auxotroph dikaryon strain is obtained by means of mon-mon cross breeding.The method is simple, cost is low, a strain obtained through a conventional strain separation method cannot grow or reproduce normally, and the lentinus edodes strain can be effectively protected through the method.

Description

Technical field [0001] The invention belongs to the field of mushroom strain protection, and particularly relates to a method for protecting mushroom strains by using uracil auxotrophy. Background technique [0002] Lentinulaedodes is one of the important cultivated edible fungi in our country and has important edible and medicinal value. The cultivation and production of shiitake mushrooms must depend on the strains to a large extent. The selection and breeding of strains can only be harvested by long-term accumulation, which has invested a lot of manpower, material resources and time costs. However, because shiitake mushrooms reproduce vegetatively, the production process of the strain is reproducible, making it easy to be counterfeited. [0003] The auxotrophic strain loses the ability to synthesize a certain substance due to genetic mutations, and cannot grow normally in the basic medium, and must be supplemented with certain substances to grow normally. If it is not clear w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/04C12N1/14C12R1/645
CPCC12N1/14C12N15/04
Inventor 周陈力鲍大鹏李燕万佳宁奚莉萍冯爱萍茅文俊汪滢王莹高英女唐利华杨瑞恒龚明
Owner SHANGHAI ACAD OF AGRI SCI
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