Simple and efficient plastid transformation system for the liverwort Marchantia polymorpha L. suspension-culture cells

被引:27
作者
Chiyoda, Shota
Linley, Philip J.
Yamato, Katsuyuki T.
Fukuzawa, Hideya
Yokota, Akiho
Kohchi, Takayuki [1 ]
机构
[1] Kyoto Univ, Grad Sch Biostudies, Div Integrated Life Sci, Kyoto 6068502, Japan
[2] Nara Inst Sci & Technol, Grad Sch Biol Sci, Nara 6300192, Japan
关键词
plastid transformation; Marchantia polymorpha; homoplasmy; bryophyte; suspension-culture; STABLE CHLOROPLAST TRANSFORMATION; HIGH-VELOCITY MICROPROJECTILES; LACTUCA-SATIVA L; HIGHER-PLANTS; GENE ORGANIZATION; TOBACCO; GENOME; PROTOPLASTS; MARKER; DNA;
D O I
10.1007/s11248-006-9027-1
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:41 / 49
页数:9
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