关键词: 糙皮侧耳, 疏水蛋白, RNAi, 根癌农杆菌介导

Abstract:

RNAi vector for Pleurotus ostreatus hydrophobin gene, Po.hyd, was constructed and transformants were
obtained by Agrobacterium tumefaciens mediated transformation. Po.hyd was cloned by PCR method using the Golden
Gate technology to build the interference vector p1302-GG-ihyd, which had a neck ring structure. Finally, this vector was
transferred into A. tumefaciens GV3101 under the induction of acetosyringone (AS), the young mycelia were mediated by
A. tumefaciens, and the transformants were detected and identified by PCR amplification, fluorescence microscopy, real-time
PCR and Southern blot technique. One genetically stable P. ostreatus transformant was obtained, which was hygromycin
resistant. The green fluorescence signal could be detected under a fluorescence microscope, and the expression of Po.hyd
was 43% compared with the wild type strain. The exogenous DNA was integrated into the genome in a single copy. This
result will lay the preliminary foundation for further studies on the function of Po.hyd.

Key words: Pleurotus ostreatus, hydrophobin, RNAi, Agrobacterium tumefaciens mediated transformation