ISSN 2736-1756
Advanced Journal of Microbiology Research Vol. 2015
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2015 International Scholars Journals
Full Length Research Paper
An efficient protoplast isolation and regeneration system in Coprinus comatus
Linhui Li1*, Qingyu Yin1, Xiaohong Liu1 and Hong Yang2
1Key Laboratory of Southwest China Wildlife Resources Conservation, China West Normal University, Nanchong City 637009, P. R. China.
2Department of Biochemistry, Xichang College, Xichang City 615022, P. R. China.
Accepted 24 February, 2014
Abstract
An efficient protocol for protoplast isolation and regeneration was first established in Coprinus Comatus. The highest yield of protoplasts was up to 8.9 × 106 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.
Key words: Coprinus comatus, mycelia, lywallzyme, protoplast, liberation and regeneration.