Obtaining insertion mutants in Magnaporthe grisea by Restriction Enzyme-Mediated Integration of plasmid

REMI (Restriction Enzyme-Mediated Integration) has been successfully used to generate random insertions and promote the transformation frequency in filamentous fungi. The protoplasts of M. grisea were transformed by enzyme-linearized pSCN43 or pBF101 along with the same restriction enzyme, and 450 transformants were obtained. After some kinds of phenotype were tested, two conidium mutants bearing strange (such as longer and narrower) conidia were recovered from these transformants. This method of insertion mutagenesis should be very useful for tagging, and subsequent cloning of many development genes that can be identified by the mutant phenotype.