Phellinus linteus mother strain tissue isolation transfer-tube culture method
A technology of tissue separation and cultivation method, which is applied in the field of tissue separation and transfer tube cultivation of Phellinus japonica mother species, can solve the problems of slow germination speed of mycelia, small one-time inoculation amount, high pollution rate, etc., to improve robustness, increase The effect of one inoculation amount and low contamination rate
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Embodiment 1
[0029] The method for cultivating the tissue separation of Phellinus japonica female species by transferring tubes, comprising the following steps:
[0030] (1) Cut the bulk tissue from the Phellinus fruiting body, inoculate the bulk tissue into a test tube culture medium, and cultivate it in a constant temperature dark room until yellow fluffy mycelium grows;
[0031] That is, in the sterilized inoculation box, the surface-sterilized wild Phellinus fruiting body is divided into two halves with both hands, and a small square tissue of about 2*2mm is cut out. Use an inoculation needle to transfer the bacterial flesh tissue to the center of the test tube culture medium. Among them, the utensils and the cut tissue must not be in contact with other parts of the Phellinus thallus to avoid contamination. After inoculation Place it in a constant temperature darkroom at 25°C for cultivation, and after 2 days, yellow fluffy hyphae can be seen growing from the tissue block. At this time...
Embodiment 2
[0039] The difference between this embodiment and embodiment 1 is: in the present embodiment, the components of the test tube culture medium and the expansion medium are different, and the specific composition is as follows:
[0040] 200g peeled potatoes, 20g glucose, KH 2 P0 4 2g, MgSO 4 1g, 1000ml of mulberry juice. The specific production process of mulberry juice is as follows: crush biennial wild mulberry branches with a crusher and pass through a 20-mesh sieve to obtain mulberry branch powder. Weigh 15g of mulberry branch powder and put it into 1000ml water, boil for 15 minutes, and filter it with gauze. Mulberry juice.
[0041] The mother Phellinus japonica produced in this example was stored under the condition of 4-6°C, and the vigor of the Phellinus japonica mother seed was detected respectively at the storage time of 60d, 80d, 100d, and 120d. After testing, the above-mentioned 60d, 80d At 100 d, the mycelium germination speed is even slower, and the mycelium can...
Embodiment 3
[0043] The difference between this embodiment and embodiment 1 is: in the present embodiment, the components of the test tube culture medium and the expansion medium are different, and the specific composition is as follows:
[0044] Peeled potato 190g, glucose 20g, KH 2 P0 4 2g, MgSO 4 1g, VB110mg, mulberry branch powder 15g, water 1000ml. The preparation method of the mulberry branch powder is as follows: the annual wild mulberry branch is dried until the water content is lower than 15%, then crushed with a pulverizer and passed through a 40-mesh sieve to obtain the mulberry branch powder.
[0045] The mother Phellinus japonica produced in this example was stored under the condition of 4-6°C, and the vigor of the Phellinus japonica mother seed was detected respectively at the storage time of 60d, 80d, 100d, and 120d. After testing, the above-mentioned 60d, 80d 、 At 100 days, the Phellinus japonica parent species can germinate mycelium normally; at 120 days, the Phellinus ...
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