Establishment of an Efficient Transformation System for Pleurotus Ostreatus

Overview

Journal World J Microbiol Biotechnol

Publisher Springer

Specialties Biotechnology
Microbiology

Date 2017 Nov 23

PMID 29164387

Citations 7

Authors

Min Lei

Xiangli Wu

Jinxia Zhang

Hexiang Wang

Chenyang Huang

Affiliations

Soon will be listed here.

Abstract

Pleurotus ostreatus is widely cultivated worldwide, but the lack of an efficient transformation system regarding its use restricts its genetic research. The present study developed an improved and efficient Agrobacterium tumefaciens-mediated transformation method in P. ostreatus. Four parameters were optimized to obtain the most efficient transformation method. The strain LBA4404 was the most suitable for the transformation of P. ostreatus. A bacteria-to-protoplast ratio of 100:1, an acetosyringone (AS) concentration of 0.1 mM, and 18 h of co-culture showed the best transformation efficiency. The hygromycin B phosphotransferase gene (HPH) was used as the selective marker, and EGFP was used as the reporter gene in this study. Southern blot analysis combined with EGFP fluorescence assay showed positive results, and mitotic stability assay showed that more than 75% transformants were stable after five generations. These results showed that our transformation method is effective and stable and may facilitate future genetic studies in P. ostreatus.

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