A method for protecting Pleurotus eryngii strains by using uracil auxotrophy

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

Active Publication Date: 2020-08-04
SHANGHAI ACAD OF AGRI SCI +1
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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, since Pleurotus eryngii 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 Pleurotus eryngii strains by using uracil auxotrophy
  • A method for protecting Pleurotus eryngii strains by using uracil auxotrophy
  • A method for protecting Pleurotus eryngii strains by using uracil auxotrophy

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Effect test

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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Abstract

The invention relates to a method for protection of a pleurotus eryngii strain by using uracil auxotrophy, which comprises the following steps of hybridizing a pleurotus eryngii uracil auxotrophy monokaryon NX2-0 with a pleurotus eryngii monokaryon strain, and after fruiting, performing single spore isolation to obtain a monokaryon strain; inoculating the monokaryon strain on a 5-fluororotic acidscreening medium for culture and identification, so as to obtain a uracil auxotrophy monokaryon strain, and then performing mon-mon crossing to obtain a uracil auxotrophy dikaryon strain. The method disclosed by the invention has the advantages of simpleness and low cost, and the strain obtained by the conventional strain isolation method cannot grow and multiply normally, and the pleurotus eryngii strain can be effectively protected.

Description

technical field [0001] The invention belongs to the technical field of Pleurotus eryngii strain protection, in particular to a method for protecting Pleurotus eryngii strains by using uracil auxotrophs. Background technique [0002] Pleurotus eryngii (Pleurotus eryngii), also known as Pleurotus eryngii, is a large-scale toadstool with high quality. It is high in protein, rich in dietary fiber, and has high edible value. Some tumors also have a certain inhibitory effect, which can enhance the body's immunity and reduce the risk of atherosclerosis. The development of the cultivation and production of Pleurotus eryngii must depend on strain 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, since Pleurotus eryngii can be reproduced asexually, its strain production process is reproducible, making it easy to be counterfeited. [0003] Auxotrop...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/04C12R1/645
Inventor 鲍大鹏周陈力万佳宁卢绪志余养朝张光忠茅文俊吴莹莹杨瑞恒李燕王莹汪滢唐利华高英女龚明
Owner SHANGHAI ACAD OF AGRI SCI
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