A high-yield and high-quality cultivated strain of Cordyceps guangdong and its identification primers and identification methods
A technology of Cordyceps sinensis and strains in Guangdong is applied in the field of high-yield and high-quality Guangdong Cordyceps sinensis cultivation strains and identification primers, which can solve the problems of harming consumers' rights and interests, misleading consumers, etc., and achieves the effects of good fruiting body traits, protection of rights and interests, and short growth cycle.
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Embodiment 1
[0023] Example 1: Mycelium growth and characteristics of Guangdong Cordyceps strain GDGM 30035
[0024] Recipe for SPDA (Sprout Potato Dextrose Agar Medium): Cut peeled 200g potatoes into a volume of 1cm 3 Mix small pieces of soybean sprouts with 100g of fresh soybean sprouts, add 800mL of distilled water to cook for 12-15min, filter through 8 layers of gauze, add 20g of glucose and 15g of agar to the filtrate, add water to make up to 1000mL, sterilize at 115°C for 20min, and set aside.
[0025] Take the mycelial block of Cordyceps guangdong strain GDGM 30035 with a diameter of about 8mm and inoculate it to the central position of the medium on the SPDA plate. And the mycelial blocks inoculated with Cordyceps guangdong strains GDGM 33006 and GDGM 43414 were used as controls. During the growth stage of mycelia, the culture should be protected from light. The temperature of the cultivation room should be controlled at 24°C, the relative humidity of the air should be controlled ...
Embodiment 2
[0026]Example 2: Fruiting body growth and characteristics of Cordyceps guangdong strain GDGM 30035
[0027] After the mycelium is overgrown with the culture medium, increase the scattered light irradiation and use LED light strips for supplementary light. The light quality can be white light or blue light, the light intensity is 220lux, the light time is 12 hours a day, and the relative air humidity is kept at 75%. After 3-4 days, the hyphae turned from white to dark green, and after 9 days, the primordia appeared. Ventilate for the first time when the primordium emerges, that is, puncture the sealing film on the cultivation container to ventilate, and the CO in the cultivation room 2 The concentration is controlled at 0.4% v / v, CO in the container 2 The concentration was controlled at 4% v / v. The temperature is controlled at 23°C and the relative air humidity is 85%. Regularly turn on the internal circulation ventilation and external circulation fan every day to keep the a...
Embodiment 3
[0029] Example 3: Molecular biological characteristics of Cordyceps guangdong strain GDGM 30035
[0030] Genomic DNA extraction of fruiting bodies of Cordyceps guangdong strain GDGM 30035: Weigh 0.15 g of fruiting bodies of Cordyceps guangdong strain GDGM30035 and place them in a sterile EP tube, pour liquid nitrogen into them and grind them quickly. HiPure Fungal DNA Mini Kit from Magen Company ( Guangzhou Meiji Biotechnology Co., Ltd. (No. D3171-03) extracted genomic DNA from Cordyceps guangdong strain GDGM 30035.
[0031] Using ISSR nucleic acid molecule primer UBC822, the nucleotide sequence is (TC) 8 A, ie: 5'-TCTCTCTCTCTCTCTCCA-3' (synthesized by Shanghai Sangon Bioengineering Co., Ltd.), PCR amplification was performed using Genomic DNA of Cordyceps guangdongensis strain GDGM 30035 as a template. The details are as follows: PCR reaction was carried out by using Premix TaqTM (TaKaRa TaqTM Version 2.0) kit (Dalian Baosheng Biological Takara, No.: R004A). The reaction sy...
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