Simple and efficient plastid transformation system for the liverwort Marchantia polymorpha L. suspension-culture cells.
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.
Chiyoda
Shota
S
Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8502, Japan.
Linley
Philip J
PJ
Yamato
Katsuyuki T
KT
Fukuzawa
Hideya
H
Yokota
Akiho
A
Kohchi
Takayuki
T
D001483:Base Sequence; D018929:Cell Culture Techniques; D018744:DNA, Plant / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D017343:Genes, Plant; D005822:Genetic Vectors; D047008:Marchantia / Q000166:cytology / Q000187:drug effects / Q000235:genetics*; D030821:Plants, Genetically Modified; D018087:Plastids / Q000235:genetics*; D000198:Spectinomycin / Q000494:pharmacology; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic
D018744:DNA, Plant; D000198:Spectinomycin