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Method for breeding high-temperature resistant strain of stropharia rugosoannulata

A technology of high-temperature resistant strains and Oleurotus giganta, applied in the methods of using microorganisms, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of mycelium not resistant to high temperature, and the fruit bodies are easy to open umbrellas, etc.

Active Publication Date: 2018-11-06
SHANDONG AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0004] In view of the above-mentioned prior art, in order to solve the problem that the mycelia of early sowing in early autumn are not resistant to high temperature, and the fruiting bodies of late autumn sowing in the next year are easy to open umbrellas, the purpose of the present invention is to provide a selection of high temperature resistant bacterial strains of Stropharia stropharcoides. Breeding method, the present invention selects and breeds high-temperature-resistant strains of Stropharia stropharia through mycelium high-temperature-resistant domestication and protoplast breeding, and verifies its stability and high-temperature-resistant quality in spring warm sheds, winter warm sheds, and forest environments, and screens Produce high-yield and high-quality high-temperature-resistant strains

Method used

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  • Method for breeding high-temperature resistant strain of stropharia rugosoannulata
  • Method for breeding high-temperature resistant strain of stropharia rugosoannulata
  • Method for breeding high-temperature resistant strain of stropharia rugosoannulata

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preparation example Construction

[0038] The preparation and regeneration of protoplasts is the basis of protoplast breeding. The treatment conditions of lytic enzyme are the key to the preparation of protoplasts. If the concentration of lytic enzyme is too low, the release efficiency of protoplasts will be low; if the concentration of enzyme is too high, the bacteria will The silk cell wall is completely dissolved, and the cells are damaged and easily ruptured, which affects the regeneration ability of the protoplast. It has been found through many experiments that the optimal concentration of lysozyme for protoplast preparation is 1.5% (mass concentration).

[0039] The enzymatic hydrolysis temperature has a great influence on the yield of protoplasts, mainly in two aspects. On the one hand, with the increase of temperature, the reaction speed is accelerated, and the release of protoplasts is continuously increasing; on the other hand, the protein is gradually denatured as the temperature increases. The react...

Embodiment 1

[0051] Embodiment 1: Breeding of Stropharia stropharia high temperature resistant strain

[0052] 1. Collect large stropharia from major cultivation areas of Shanghai Academy of Agricultural Sciences, Shandong Academy of Agricultural Sciences, Taian Academy of Agricultural Sciences, Sanming Fungi Research Institute, Sichuan Soil and Fertilizer Research Institute, Dalian in Liaoning, Suihua in Heilongjiang, Yuxi in Yunnan, and Jining in Shandong. A total of 23 Stropharia strains or fruiting bodies were obtained (Table 1). Among them, Mingda 128 (SM) and Stropharia 1 (TF) were the varieties certified by the state in 2008.

[0053] Table 1: Strain names and sources

[0054]

[0055] 2. Analysis of strain genetic diversity and kinship:

[0056] 2.1 Method:

[0057] 2.1.1 Strain activation, culture and DNA extraction

[0058] The collected strains and fruit bodies collected from the cultivation site were separated and purified by tissue culture, and then stored in a 4°C refr...

Embodiment 2

[0089] Embodiment 2: Breeding of Stropharia stropharia high temperature resistant strain

[0090] The difference with Example 1 is: after the protoplasts are prepared, the protoplasts are diluted to 10 5 cells / mL, gradient mutagenesis was carried out at a distance of 30cm with a stable 15W ultraviolet lamp, and the mutagenesis time was set to 30s. Then spread the protoplasts irradiated by ultraviolet rays on the regeneration medium, and recover at 26°C for 2 days, then adjust the temperature to 34°C to continue culturing. Eliminate the monokaryotic strains among them, and number the remaining strains according to 1st to 20th, and cultivate them at 26°C for 5 days, then adjust the temperature to 34°C, continue to cultivate for 5 days, and then adjust the temperature to 26°C to resume the culture for 5 days. The growth rate of each strain was recorded and compared, and 3 strains with fast mycelium growth, good growth and low heterogeneity after being cultured at 34°C were selec...

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Abstract

The invention discloses a method for breeding a high-temperature resistant strain of stropharia rugosoannulata. The method comprises the following steps: (1) collecting stropharia rugosoannulata strains or sporocarp, carrying out genetic diversity and genetic relationship analysis, rejecting strains in close genetic relationships, and collecting rest strains as test strains; (2) carrying out hyphahigh-temperature resistant progressive circulation domestication on the test strains, and screening high-temperature domesticated strains of stropharia rugosoannulata; (3) inoculating the high-temperature domesticated strains of the stropharia rugosoannulata into a liquid medium, leaving to stand and culture, collecting hyphae, and carrying out enzymolysis with lywallzyme so as to obtain protoplast; (4) carrying out high-temperature regeneration testing on the protoplast, and screening a high-temperature resistant strain. A high-quality high-yield high-temperature resistant strain is screenedby using the method, the problems that early sowing hyphae in early autumn are not capable of enduring high temperatures and the cap of sporocarp is liable to open at the end of spring and at the beginning of summer if seeds are sown in late autumn in production can be solved, and high-quality high-yield strains can be provided for cultivation in early autumn and harvesting in early winter and prolonging of mushroom harvesting periods in spring in the Eastern China.

Description

technical field [0001] The invention relates to the technical field of fungal breeding, in particular to a method for breeding high-temperature-resistant strains of Stropharia grandis. Background technique [0002] Stropharia rugosoannulata is an important edible and medicinal fungus. It is delicious in taste, rich in nutrition, and has health care and medicinal values ​​such as enhancing human immunity and lowering blood sugar. Stropharia is a medium-low temperature edible fungus. Its mycelial growth temperature range is 5-32°C, and the temperature range for fruiting body formation and growth is 10-20°C. When the temperature rises to 20°C, the fruiting body is easy to open. , the quality deteriorates, and the original base does not differentiate at 25°C. Therefore, in the open-air cultivation in East China, if sowing in August-September, the product quality is good when fruiting in November-December, and it is durable to storage, but after sowing, the mycelia meet 30 High ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/36C12N1/14C12N13/00C12R1/645
CPCC12N1/14C12N1/36C12N13/00
Inventor 姜淑霞朱静娴孟丽王庆佶李丽君任纪帆
Owner SHANDONG AGRICULTURAL UNIVERSITY
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