Simple and efficient plastid transformation system for the liverwort Marchantia polymorpha L. suspension-culture cells |
| |
Authors: | Shota Chiyoda Philip J. Linley Katsuyuki T. Yamato Hideya Fukuzawa Akiho Yokota Takayuki Kohchi |
| |
Affiliation: | (1) Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan;(2) Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan |
| |
Abstract: | We have established a simple and efficient plastid transformation system for liverwort, Marchantia polymorpha L., suspension-culture cells, which are homogenous, chloroplast-rich and␣rapidly growing. Plasmid pCS31 was constructed to integrate an aadA expression cassette for spectinomycin-resistance into the trnI–trnA intergenic region of the liverwort plastid DNA by homologous recombination. Liverwort suspension-culture cells were bombarded with pCS31-coated gold projectiles and selected on a medium containing spectinomycin. Plastid transformants were reproducibly isolated from the obtained spectinomycin-resistant calli. Selection on a sucrose-free medium greatly improved the efficiency of selection of plastid transformants. Homoplasmic plastid transformant lines were established by␣successive subculturing for 14 weeks or longer on the spectinomycin-containing medium. The plastid transformation system of liverwort suspension-culture cells should facilitate the investigation of the fundamental genetic systems of plastid DNA, such as replication. |
| |
Keywords: | Plastid transformation Marchantia polymorpha Homoplasmy Bryophyte Suspension-culture |
本文献已被 PubMed SpringerLink 等数据库收录! |
|