Agrobacterium tumefaciens-mediated transformation of Botryosphaeria dothidea

作者：Chen, Liang; Wang, Qun; Chen, Hua; Sun, Gengwu; Liu, Huixiang^*; Wang, Hongkai^*

来源：WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY, 2016, 32(7): 106.

DOI：10.1007/s11274-016-2045-0

• Botryosphaeria dothidea
• ATMT
• T-DNA
• Hygromycin B
• hph

Botryosphaeria dothidea is a severe causal agent of die-back and cankers of many woody plants and causes great losses in many regions. The pathogenic mechanism of this pathogen has not been well explored due to lack of mutants and genetic information. In this study, we developed an Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for B. dothidea protoplasts using vector pBHt2 containing the hph gene as a selection marker under the control of trp C promoter. Using this protocol we successfully generated the B. dothidea transformants with efficiency about 23 transformants per 105 protoplasts. This is the first report of genetic transformation of B. dothidea via ATMT and this protocol provides an effective tool for B. dothidea genome manipulation, gene identification and functional analysis.

• 出版日期2016-7
• 单位浙江工商大学; 山东农业大学; 浙江大学