title: Plastid transformation of sporelings and suspension-cultured cells from the liverwort Marchantia polymorpha L.
abstract: We describe simple and efficient plastid transformation methods for suspension-cultured cells and sporelings of the liverwort, Marchantia polymorpha L. Use of rapidly proliferating cells such as suspension-cultured cells and sporelings, which are immature thalli developing from spores, as targets made plastid transformation by particle bombardment efficient. Selection on a sucrose-free medium and linearization of the transformation vector significantly improved the recovery rate of plastid transformants. With the methods described here, a few plastid transformants are obtained from a single bombardment of sporelings, while more efficient plastid transformation is expected in suspension-cultured cells, ~60 transformants from a single bombardment. Homoplasmic transformants of thalli are obtained immediately after primary selection, whereas homoplasmic transformants from suspension-cultured cells are obtained after 12-16 weeks of repeated subculture.
authors.affiliation: Graduate School of Biostudies, Kyoto University, Kyoto, Japan.
authors.forename: Shota
authors.initials: S
authors.lastname: Chiyoda
authors.forename: Katsuyuki T
authors.initials: KT
authors.lastname: Yamato
authors.forename: Takayuki
authors.initials: T
authors.lastname: Kohchi
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019470:Biolistics / Q000379:methods*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D002736:Chloroplasts / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D059765:Homologous Recombination; D047008:Marchantia / Q000235:genetics*; D009713:Nucleotidyltransferases / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D000198:Spectinomycin / Q000494:pharmacology; D013170:Spores / Q000254:growth & development; D014170:Transformation, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D018744:DNA, Plant; D000198:Spectinomycin; D009713:Nucleotidyltransferases; C020565:gentamicin 2''-nucleotidyltransferase