Screening and identification of insertion mutants from Bipolaris eleusines by mutagenesis based on restriction enzyme-mediated integration.

摘要: Ophiobolin A is sesterterpenoid-type phytotoxin and may be an important candidate for development of new crop protection pharmaceutical products. The restriction enzyme-mediated integration (REMI) method was used to introduce the plasmid pSH75 into ophiobolin A-producing filamentous fungus Bipolaris eleusines. total 323 stable transformants were obtained, all which capable growing on potato-dextrose agar medium containing 200 μg mL−1 hygromycin B. transformation frequency about 4–5 transformants μg−1 DNA. An ophibolin A-deficient transformant (B014) assessed presence hph gene in this confirmed by PCR. cell-free cultural filtrates showed significantly less inhibition mycelial growth fungal pathogen Rhizoctoni solani but little effect barnyard grass as opposed that wild-type B. eleusines. There no detectable amount B014 samples measured with HPLC. This research suggests REMI a potential approach improving production B. eleusines via genetic engineering upregulate certain genes responsible desired biosynthetic pathways.