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

Kimitsune Ishizaki; Ryuichi Nishihama; Minoru Ueda; Keisuke Inoue; Sakiko Ishida; Yoshiki Nishimura; Toshiharu Shikanai; Takayuki Kohchi

doi:10.1371/journal.pone.0138876

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.

Authors

Kimitsune Ishizaki¹, Ryuichi Nishihama², Minoru Ueda³, Keisuke Inoue², Sakiko Ishida², Yoshiki Nishimura³, Toshiharu Shikanai³, Takayuki Kohchi²

Affiliations

¹ Graduate School of Biostudies, Kyoto University, Kyoto, Japan; Graduate School of Science, Kobe University, Kobe, Japan.
² Graduate School of Biostudies, Kyoto University, Kyoto, Japan.
³ Department of Botany, Graduate School of Science, Kyoto University, Kyoto, Japan.

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

Grants and funding

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.