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.
CITATION STYLE
Chiyoda, S., Yamato, K. T., & Kohchi, T. (2014). Plastid transformation of sporelings and suspension- Cultured cells from the liverwort marchantia polymorpha L. Methods in Molecular Biology, 1132, 439–447. https://doi.org/10.1007/978-1-62703-995-6_30
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