Wolfiporia cocos YX1 and culture media and cultivation method thereof
A cultivation method and technology of original seed culture medium are applied in the field of Poria cocos YX1 and its culture medium, and can solve the problem of difficulty in realizing standardization, normalization, large-scale industrial cultivation, cultivation time limitation of Poria cultivation methods, wild resource collection and The protection power is not strong and other problems, to promote the growth of Poria mycelium, solve the shortage of raw material supply, and achieve the effect of good quality of Poria cocos
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Embodiment 1
[0053] Screening of Wolfiporia cocos YX1
[0054] From the pine forest area of Qingzhai Village, Zhongguan Town, Yuexi County, Anqing City, Anhui Province (30 ° 85' north latitude, 116 ° 35' east longitude), under the decayed pine stump, the dormant sclerotium of the brown rot fungus Poria cocos was collected, and its Mycelium, and then inoculated into different formulations of the mother seed culture medium in the ultra-clean workbench, cultured, and observed the growth of Poria cocos mycelium; then the well-screened mother species were inoculated into different formulations of the original seed culture medium , cultured, observed the growth of Poria cocos hyphae, and screened good original species; then continued to transfer for two generations to obtain pure Poria cocos mycelium, and preserved the above-mentioned Poria cocos (Wolfiporia cocos) YX1 in the Chinese Type Culture Collection in Wuhan Center (CCTCC), the deposit number is CCTCC NO: M 2021434.
Embodiment 2
[0056] The Poria cocos (Wolfiporia cocos) YX1 obtained in Example 1 was subjected to morphological identification of Poria cocos hyphae.
[0057] 1. Specific method: Inoculate the Poria cocos (Wolfiporia cocos) YX1 obtained in Example 1 in a petri dish equipped with medium, cultivate for 4-5 days, then insert the glass slide into the medium at an angle of 45°, and cultivate for 12 hours Take out the glass slide, put the cover glass on the optical microscope and electron microscope, observe the macroscopic and microscopic growth characteristics such as the formation, color, structure and whether there is separation of Poria cocos hyphae under the field of view of 10 times or 20 times of the objective lens, Such as figure 1 as shown, figure 1 It is the culture characteristic picture of Poria cocos (Wolfiporia cocos) YX1 mycelia, wherein, a is the front of the culture dish, b is the back of the culture dish, c is the picture of 10 times of objective lens, and d is the picture of...
Embodiment 3
[0063] Molecular biological identification of Wolfiporia cocos YX1 obtained in Example 1 was carried out.
[0064] 1. Extract the total genomic DNA of Wolfiporia cocos (Wolfiporia cocos) YX1: put 10-30 mg of mycelium or sclerotia of fresh Wolfiporia cocos YX1 into a centrifuge tube, and grind it fully into a sample preparation system cell disruptor. Powder until the wall is broken, crushed at 5000rpm for 30 seconds, and repeat 2-3 times to obtain a broken wall sample; then follow the CTAB method or UNIQ-10 column type fungal genome extraction kit (manufacturer: Sangon Bioengineering (Shanghai) Co., Ltd., No.: B511375) method to extract the total genomic DNA sample, and store it in a -20°C refrigerator for later use;
[0065] 2. Obtain the rDNA sequence, 28S sequence, and EF sequence of Wolfiporia cocos YX1: Use the total genomic DNA obtained in step 1 as a template, and then perform polymerase chain PCR amplification. The total system is 25 μl, using three pairs of Primers IT...
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