An Efficient PEG/CaCl₂-Mediated Transformation Approach for the Medicinal Fungus Wolfiporia cocos

J Microbiol Biotechnol. 2015 Sep;25(9):1528-31. doi: 10.4014/jmb.1501.01053.

Abstract

Sclerotia of Wolfiporia cocos are of medicinal and culinary value. The genes and molecular mechanisms involved in W. cocos sclerotial formation are poorly investigated because of the lack of a suitable and reproducible transformation system for W. cocos. In this study, a PEG/ CaCl₂-mediated genetic transformation system for W. cocos was developed. The promoter Pgpd from Ganoderma lucidum effectively drove expression of the hygromycin B phosphotransferase gene in W. cocos, and approximately 30 transformants were obtained per 10 μg DNA when the protoplast suspension density was 10(6) protoplasts/ml. However, no transformants were obtained under the regulation of the PtrpC promoter from Aspergillus nidulans.

Keywords: Genetic transformation; Wolfiporia cocos; medicinal and culinary value; promoter.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antifungal Agents / metabolism
  • Aspergillus nidulans / genetics
  • Calcium Chloride / metabolism*
  • Coriolaceae / drug effects*
  • Coriolaceae / genetics*
  • Gene Transfer Techniques*
  • Genetics, Microbial / methods*
  • Hygromycin B / metabolism
  • Phosphotransferases (Alcohol Group Acceptor) / metabolism
  • Polyethylene Glycols / metabolism*
  • Promoter Regions, Genetic
  • Reishi / genetics
  • Selection, Genetic
  • Transformation, Genetic*

Substances

  • Antifungal Agents
  • Polyethylene Glycols
  • Hygromycin B
  • Phosphotransferases (Alcohol Group Acceptor)
  • hygromycin-B kinase
  • Calcium Chloride