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A method for protecting mushroom strains by using uracil auxotroph

An auxotrophic, uracil technology, applied in the directions of microorganism-based methods, biochemical equipment and methods, fungi, etc., can solve problems such as being easily counterfeited, and achieve the effect of low cost and simple method

Active Publication Date: 2018-11-20
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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  • A method for protecting mushroom strains by using uracil auxotroph
  • A method for protecting mushroom strains by using uracil auxotroph
  • A method for protecting mushroom strains by using uracil auxotroph

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Obtaining Monospores with Uracil Auxotrophy in Lentinus edodes

[0035] The auxotrophic strain 423-9 of Lentinus edodes was crossed with monokaryotic strain W66 1 of Lentinus edodes to obtain dikaryotic strain Z2 for fruiting. After fruiting, select two single fruiting bodies with complete mushroom shape, large size, thick flesh, and no pests and diseases, and use a flat plate to collect naturally ejected spores in a sterile bell jar 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. Germinated spores are generally visible within 2 weeks. 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 association...

Embodiment 2

[0037] Preparation of uracil auxotrophic dikaryotic strain from Lentinus edodes

[0038] The monokaryotic hyphae obtained from the germination of the uracil auxotrophic monospore strain were hybridized on the MMU medium. After about two weeks of cultivation, the mycelia of the parents were found to have fused, and the hyphae at the fusion point were picked out for microscopic examination. The mycelia with lock-like joints were uracil auxotrophic dikaryotic mycelia. (UH9 is obtained by crossing Z2-14 and Z2-97, and UH21 is obtained by crossing Z2-18 and Z2-97. Z2-14, Z2-18, and Z2-97 are collection sheets of fruiting bodies obtained by crossing W66 1 and 423-9 spores obtained).

[0039] The UH9 preservation number is: CGMCC NO.11916, and the taxonomy name: Lentinula edodes is preserved on January 29, 2016. Deposit unit: CGMCC, China General Microorganism Culture Collection and Management Center. Preservation address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, B...

Embodiment 3

[0042] Validation of 5'-fluoroorotic acid in uracil auxotrophic dikaryotic strain

[0043] The obtained uracil auxotrophic dikaryotic strains of Lentinus edodes 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 The indicated uracil auxotrophic dikaryotic 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 protection of shiitake mushroom strains, in particular to a method for protecting shiitake mushroom strains by using uracil auxotrophs. Background technique [0002] Mushroom (Lentinula edodes) is one of the important cultivated edible fungi in my country, which has important edible and medicinal value. The development of the cultivation and production of shiitake mushrooms must depend on the strains 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 shiitake mushrooms reproduce asexually, the production process of their strains 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 sub...

Claims

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Application Information

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Patent Type & Authority Patents(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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