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

PLoS One. 2015 Sep 25;10(9):e0138876. doi: 10.1371/journal.pone.0138876. eCollection 2015.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetolactate Synthase / genetics
  • Acetolactate Synthase / metabolism*
  • Acetyltransferases / genetics
  • Acetyltransferases / metabolism*
  • Agrobacterium tumefaciens / enzymology
  • Agrobacterium tumefaciens / genetics*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Genetic Markers / genetics
  • Genetic Vectors / genetics
  • Gentamicins / pharmacology
  • Kanamycin Kinase / genetics
  • Kanamycin Kinase / metabolism*
  • Marchantia / drug effects
  • Marchantia / genetics*
  • Marchantia / growth & development
  • Plants, Genetically Modified / growth & development
  • Sulfonamides / pharmacology
  • Transfection
  • Triazines / pharmacology

Substances

  • Bacterial Proteins
  • Genetic Markers
  • Gentamicins
  • Sulfonamides
  • Triazines
  • antibiotic G 418
  • Acetolactate Synthase
  • Acetyltransferases
  • gentamicin 3-acetyltransferase
  • Kanamycin Kinase
  • chlorsulfuron

Grant support

MEXT KAKENHI Grants-in-Aid for Scientific Research on Priority Areas (23012025 to T.K.) and for Scientific Research on Innovative Areas (25113009 to T.K., 25119711 and 15H01233 to K.Is.); by JSPS KAKENHI Grants-in-Aid for Scientific Research (C) (24580140 to K.Is., 24570048 to R.N.) and for Challenging Exploratory Research (26650095 to T.K.); by a research fellowship from the JSPS for young scientists (21-764 to M.U., 24-7049 to K.In). URL for Kakenhi: http://www.jsps.go.jp/english/e-grants/index.html. This work was also funded by the Asahi Glass Foundation (http://www.af-info.or.jp/en/index.html) and the SUNTORY Foundation for Life Sciences (http://www.sunbor.or.jp/egs/index.html) to K.Is. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.