Optimization of Agrobacterium tumefaciens-mediated transformation method of oleaginous filamentous fungus Mortierella alpina on co-cultivation materials choice

Shunxian Wang; Haiqin Chen; Yuzhuo Wang; Xin Tang; Hao Zhang; Wei Chen; Yong Q Chen

doi:10.1016/j.mimet.2018.08.002

Optimization of Agrobacterium tumefaciens-mediated transformation method of oleaginous filamentous fungus Mortierella alpina on co-cultivation materials choice

J Microbiol Methods. 2018 Sep:152:179-185. doi: 10.1016/j.mimet.2018.08.002. Epub 2018 Aug 7.

Authors

Shunxian Wang¹, Haiqin Chen², Yuzhuo Wang³, Xin Tang⁴, Hao Zhang⁵, Wei Chen⁶, Yong Q Chen⁷

Affiliations

¹ State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China.. Electronic address: 7140112027@vip.jiangnan.edu.cn.
² State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China.. Electronic address: haiqinchen@jiangnan.edu.cn.
³ State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China.. Electronic address: 61701120284@stu.jiangnan.edu.cn.
⁴ State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China.. Electronic address: xintang@jiangnan.edu.cn.
⁵ State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China.. Electronic address: zhanghao@jiangnan.edu.cn.
⁶ State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China.; National Engineering Research Center for Functional Food, Jiangnan University, Wuxi, Jiangsu 214122, China; Beijing Innovation Centre of Food Nutrition and Human Health, Beijing Technology and Business University (BTBU), Beijing 100048, China. Electronic address: weichen@jiangnan.edu.cn.
⁷ State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China.; National Engineering Research Center for Functional Food, Jiangnan University, Wuxi, Jiangsu 214122, China; Department of Cancer Biology, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA. Electronic address: yqchen@jiangnan.edu.cn.

PMID: 30096341
DOI: 10.1016/j.mimet.2018.08.002

Abstract

The Agrobacterium tumefaciens-mediated transformation (ATMT) method is commonly applied in the oleaginous filamentous fungus Mortierella alpina. During the ATMT process, the spores of M. alpina have traditionally been used as a co-cultivation material, but their long spore-producing cycle and low sporulation rate make the transformation process tedious. This study explores the use of germinating spores, mycelium and single solid colonies of uracil auxotrophic M. alpina CCFM501 as a co-cultivation material with A. tumefaciens AGL1. The results show that A. tumefaciens AGL1 can successfully transform the germinating spores, mycelium and single solid colonies of M. alpina. In addition, the transformation rate of the germinating spores was 50% higher than that of the fresh spores. Due to its concise preparation process, the mycelium was chosen as a co-cultivation material for two plasmids of different lengths and proven to be an efficient co-cultivation material for M. alpina.

Keywords: Agrobacterium tumefaciens-mediated transformation method; Germinating spores; M. alpina; Mycelium; Single solid colonies.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Agrobacterium tumefaciens / genetics*
Coculture Techniques / methods*
Culture Media
Genetic Engineering / methods*
Mortierella / genetics*
Mycelium / growth & development
Mycelium / metabolism
Plasmids
Spores, Fungal / genetics
Spores, Fungal / growth & development
Transformation, Genetic*

Substances

Culture Media