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We previously reported  Agrobacterium -mediated transformation methods for the liverwort  Marchantia polymorpha  using the hygromycin phosphotransferase gene as a marker for selection with hygromycin. In this study, we developed three additional markers for  M .  polymorpha  transformation: the gentamicin 3'-acetyltransferase gene for selection with gentamicin; a mutated acetolactate synthase gene for selection with chlorsulfuron; and the neomycin phosphotransferase II gene for selection with G418. Based on these four marker genes, we have constructed a series of Gateway binary vectors designed for transgenic experiments on  M .  polymorpha . The 35S promoter from cauliflower mosaic virus and endogenous promoters for constitutive and heat-inducible expression were used to create these vectors. The reporters and tags used were Citrine, 3×Citrine, Citrine-NLS, TagRFP, tdTomato, tdTomato-NLS, GR, SRDX, SRDX-GR, GUS, ELuc(PEST), and 3×FLAG. These vectors, designated as the pMpGWB series, will facilitate molecular genetic analyses of the emerging model plant  M .  polymorpha .

Development of Gateway Binary Vector Series with Four Different Selection Markers for the Liverwort Marchantia polymorpha