Agrobacterium tumefaciens-mediated transformation of Botryosphaeria dothidea.
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 10(5) 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.
Shandong Research Center for Forestry Harmful Biological Control Engineering and Technology, College of Plant Protection, Shandong Agricultural University, Tai'an, 271018, People's Republic of China.|Zhejiang Gongshang University, Hangzhou, 310018, People's Republic of China.
Liang
L
Chen
Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, 310058, People's Republic of China.
Qun
Q
Wang
Chongqing Vocational Institute of Tourism, Chongqing, 409099, People's Republic of China.
Hua
H
Chen
Shandong Research Center for Forestry Harmful Biological Control Engineering and Technology, College of Plant Protection, Shandong Agricultural University, Tai'an, 271018, People's Republic of China.
Gengwu
G
Sun
Shandong Research Center for Forestry Harmful Biological Control Engineering and Technology, College of Plant Protection, Shandong Agricultural University, Tai'an, 271018, People's Republic of China. hxliu@sdau.edu.cn.
Huixiang
H
Liu
State Key Laboratory for Rice Biology, Biotechnology Institute, Zhejiang University, Hangzhou, 310058, People's Republic of China. hkwang@zju.edu.cn.
Hongkai
H
Wang
D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006921:Hygromycin B / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D011401:Promoter Regions, Genetic; D011523:Protoplasts; D004718:Saccharomycetales / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D004271:DNA, Fungal; C036483:T-DNA; D006921:Hygromycin B
ATMT; Botryosphaeria dothidea; Hygromycin B; T-DNA; hph