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A kind of primer and method for rapid screening of high temperature resistant strain of Agaricus bisporus w192 strain
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A technology for high temperature resistant strains, Agaricus bisporus, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as limiting Agaricus bisporus
Active Publication Date: 2022-08-02
SHANGHAI ACAD OF AGRI SCI
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Problems solved by technology
Therefore, the lack of industrial strains of Agaricus bisporus in China limits the development of industrialization of Agaricus bisporus in China
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Embodiment 1
[0023] Embodiment 1: a kind of primer and method for rapid screening of thermotolerant strain of Agaricus bisporus W192 strain
[0024] Primers 4140-1F and 4140-1R were designed according to gene 4140, Seq. No. 1 and Seq. No. 2, respectively, and were used as primers for screening the thermotolerant strain of Agaricus bisporus W192 strain. The sequence of gene 4140 is Seq.No.3. According to the primers, the specific steps of the detection method are as follows:
[0026] The collected W192 hybrid strain or spore strain is cultivated on a PDA plate, and the diameter of the colony on the plate is about 2cm, pick mycelia, and use a DNA lysate to crack the picked mycelium, and the cracked liquid is used as a PCR template . DNA lysate was purchased from Shanghai Sangon Bioengineering Co., Ltd.
[0027] (2) PCR amplification
[0028] Using the mycelial lysate as the PCR template, general PCR amplification was performed using primers 414...
Embodiment 2
[0038] Embodiment 2: a kind of primer and method for rapid screening of thermostable strain of Agaricus bisporus W192 strain
[0039] 1. Collection of spore strains of strain W192
[0040] Put the fruiting bodies of the open umbrella W192 strain into the spore collector to collect the spores, spread the spores on the PDA plate, and the coated spore concentration is 10 months. 3 / mL, each plate with a diameter of 9cm was coated with 100ul of spore suspension, and placed in a 25°C incubator for cultivation.
[0041] 2. Treatment of W192 strain spore strain temperature
[0042] The germinated spore strains were treated at 40°C for 24 hours, and then transferred to 25°C for further cultivation. The spore strains that could continue to grow were used as the alternatives of the thermotolerant strains.
[0044] Pick 50 mg of the mycelium of the spore strain into a 1.5 mL centrifuge tube, add 50 ul of DNA lysis solution, and incubate ...
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Abstract
A primer and method for rapidly screening high temperature resistant strains of Agaricus bisporus W192 strain, the primer sequences are: 4140-1F: TTCCGTTCAGTGCTACC; 4140-1R: TGCAAAGAGAGCTGGTACA. The invention can quickly screen the high temperature resistant strains of the Agaricus bisporus W192 strain, and can predict the high temperature resistant characteristics of the W192 spore strain in advance for fruiting experiments, which greatly saves the time for breeding the high temperature resistant strains and promotes the breeding process. process.
Description
technical field [0001] The invention belongs to the technical field of edible fungus breeding, and relates to a primer and a method for rapidly screening high temperature resistant strains of Agaricus bisporus W192 strain. Background technique [0002] Agaricus bisporus, Chinese alias is mushroom, foreign mushroom, white mushroom. It is rich in protein, polysaccharides, vitamins, nucleotides and unsaturated fatty acids. It is not only nutritious, plump and delicious, but also low in calorie energy. It has a high health care effect and is increasingly loved by people all over the world. In foreign countries, Agaricus bisporus still dominates the consumption of edible fungi. Taking the world's second largest edible fungi production country - the United States (accounting for 16% of the world's total output) as an example, Agaricus bisporus accounts for more than 90% of its total output. In China, the consumption market of Agaricus bisporus is developing rapidly, and the sales...
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