Production of artificial hybrid lingzhi and uses thereof
A kind of Ganoderma lucidum, artificial technology, applied in the production and application field of artificial hybrid Ganoderma lucidum, can solve the problems of low seed setting rate, inability to develop nutritional mutant commercial products, hybridization failure and the like
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[0029] Artificial hybridization of Ganoderma lucidum and Ganoderma lucidum by protoplast fusion.
[0030] (a) The two parents are vegetative mycelia grown in a complete medium (CM) broth consisting of: MgSO 4 ·7H 2 O, 0.5; KH 2 PO 4 , 0.46; K 2 HPO 4 , 1; peptone, 2; D-glucose, 20 (g / L), cultured in the dark, at a temperature of 28° C. and a rotational speed of 120 rpm, and recorded the time. Actively growing mycelia were harvested using aseptic technique, filtered through a 1 mm diameter nickel mesh, and rinsed with infiltration buffer (0.6 M in sucrose solution). Then, the mycelia were cultured for 1 to 3 hours in permeabilization buffer containing 10 mg / ml of lysozyme (Guangdong Institute of Microbiology, China) to dissolve the fungal cell wall with the enzyme. The number of generated protoplasts was counted with a hemocytometer by phase contrast light microscopy.
[0031] After enough protoplasts were released, the undigested mycelia were filtered out with a syringe...
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