Agrobacterium-mediated delivery of marker genes to Phanerochaete chrysosporium mycelial pellets: a model transformation system for white-rot fungi

摘要

The lack of an efficient transformation system in filamentous fungi is one of the major problems in carrying out cloning and expression of genes to produce a value-added product at the commercial level. Therefore an attempt has been made to develop an efficient, convenient and expeditious genetic transformation system for successful transfer of transferred DNA, carrying the genes coding for GUS (beta-glucuronidase) (uidA), green fluorescent protein (gfp) and hygromycin phosphotransferase (hpt) to the nuclear genome of Phanerochaete chrysosporium, a widely studied lignin-degrading white-rot fungus. The transformation of the marker gene was confirmed through GUS, PCR and Southern-blot hybridization. Transformation with acetosyringone, in the presence of light with partial bleaching for 1 h, were found to be a better combination for fungal transformation with 48% efficiency. The agrotransformation method to transform the modified mycelial pellets offers a practical means for exploiting transgenic approaches in genetic manipulation and improvement of R chrysosporium, a model white-rot fungus. Moreover, it has the potential to overcome the technical hurdles in genetic manipulation to biotechnologically exploit many other filamentous fungi of industrial importance.