Optimization of Preparation and Regeneration Conditions of Two Kinds of Ganoderma lucidum Protoplasts
Using Ganoderma lingzhi and Ganoderma sessile strains as experimental materials,the ef-fects of mycelial age,enzymolysis time,enzymolysis temperature and centrifugal speed on the prepa-ration of Ganoderma lucidum protoplasts were compared and analyzed through single factor test and orthogonal test,the preparation conditions were optimized,and the effects of different types and con-centrations of osmotic stabilizers and different culture methods on the regeneration of Ganoderma lu-cidum protoplasts were preliminarily discussed.The results showed that the optimal conditions for the preparation of Ganoderma lingzhi protoplasts were mycelial age of 10 days,enzymolysis tem-perature of 35℃,enzymolysis for 3 hours,centrifugation at 4 000 r/min,and protoplast yield of 9.75×107/mL;Ganoderma sessile protoplasts were prepared at a mycelial age of 7 days,enzymolysis temperature of 31℃,enzymolysis for 3.5 hours,centrifugation at 3 000 r/min,and a protoplast yield of 7.48×107/mL.Ganoderma lucidum strains were regenerated with 0.5 mol/L mannitol as osmotic sta-bilizer,and the double-layer plate culture method(with an upper agar concentration of 1.1%)could obtain a high regeneration rate.
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