A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms

Yuanyuan Wang; Danyun Xu; Dongmei Liu; Xueyan Sun; Yue Chen; Lisheng Zheng; Liguo Chen; Aimin Ma

doi:10.1080/12298093.2019.1628523

A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms

Mycobiology. 2019 Jun 22;47(3):350-354. doi: 10.1080/12298093.2019.1628523. eCollection 2019.

Authors

Yuanyuan Wang¹, Danyun Xu¹, Dongmei Liu¹, Xueyan Sun¹, Yue Chen¹, Lisheng Zheng², Liguo Chen², Aimin Ma^{1

3}

Affiliations

¹ College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China.
² College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, China.
³ Key Laboratory of Agro-Microbial Resources and Utilization, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, China.

Abstract

In the post-genomic era, gene function analysis has attracted much attention. Transformation is often needed to investigate gene function. In this study, an easy, rapid, reliable, and cost-effective colony polymerase chain reaction (PCR) method for screening mushroom transformants was developed: picking up a suitable amount of transformant's tissue (1-10 μg) to 20 μl 0.25% Lywallzyme solution, and vortexing for 10 s followed by incubation at 34 °C for 15 min. Finally, 2 μl of the suspension was used as templates to perform PCR and single target bands were successfully amplified from respective transformants of Tremella fuciformis, Pleurotus ostreatus, and Pleurotus tuber-regium. This procedure could be widely employed for screening transformants in mushroom transformation experiments.

Keywords: Colony PCR; mushroom; screening; transformant.

Grants and funding

This work was supported by the grant of National Natural Science Foundation of China (NSFC) [Grant No.31572182] to Aimin Ma.