An efficient protoplast isolation and regeneration system in Coprinus comatus


Linhui Li1*, Qingyu Yin1, Xiaohong Liu1and Hong Yang2

An efficient protocol for protoplast isolation and regeneration was first established in Coprinus Comatus. The highest yield of protoplasts was up to 8.9 × 10 6 cells/ml in digestion solution containing 2.0% lywallzyme and 0.6 M KCl after incubation of four-day-old mycelia at 30°C for 3 h; among which, about 1.4% protoplasts could be regenerated into mycelia after 4 - 6 days of incubation at 25°C in CYM medium with 0.6 M mannitol as osmotic stabilizer. The results are beneficial for breeding new cultivars by the methods such as protoplast fusion, mutagenesis as well as transformation. Moreover, the stepwise procedure for protoplast liberation and regeneration could be referred in other species.

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