文章基本信息

标题：Development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus
本地全文：下载
作者：Shan Lu ; Xiaorui Shen ; Baoshan Chen 等
期刊名称：Scientific Reports
电子版ISSN：2045-2322
出版年度：2017
卷号：7
期号：1
DOI：10.1038/s41598-017-03233-7
语种：English
出版社：Springer Nature
摘要：Sporisorium scitamineum is the causative agent responsible for smut disease of sugarcane worldwide. However, lack of efficient gene manipulation system makes this fungus much behind the type model of the smut fungi in molecular biology. Here, we report the development of a CRISPR/Cas9 and T-DNA based dual vector system that allowed efficient knock-out or knock-in of a gene of interest in the S. scitamineum in a site-specific manner. By using Mfa2, a key player in the mating event in S. scitamineum as a tester gene, site-specific insertions of the introduced fragments were achieved both for Mfa2 knockout and complementation. Of particular advantage of this system is the simplicity of selection and identification for the desired transformants by using drug resistance coupled with PCR. This system greatly facilitates the gene function study in S. scitamineum, and could potentially be used for other basidiomycete fungi.