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Plant Physiol. 2016 Feb;170(2):627-41. doi: 10.1104/pp.15.01246. Epub 2015 Dec 7.

MultiSite Gateway-Compatible Cell Type-Specific Gene-Inducible System for Plants.

Author information

1
Institute of Biotechnology (R.S., X.W., S.R.Y., S.L., R.U., I.S., J.Z., A.P.M.) and Department of Biosciences, Viikki Plant Science Centre (R.S., X.W., S.R.Y., S.L., R.U., I.S., J.Z., M.W., A.P.M.), University of Helsinki, Helsinki 00014, Finland;Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, Indiana 47907 (A.M., G.M.);Molecular Genetics, Department of Biology, Utrecht University, Utrecht 3584 CH, The Netherlands (M.G., R.H., B.S.);School of Biology, Indian Institute of Science Education and Research, Thiruvananthapuram 695016, Kerala, India (K.P.);Plant Developmental Biology, Wageningen University, Wageningen 6708 PB, The Netherlands (R.H., B.S.); andDepartment of Plant Science and Landscape Architecture, University of Maryland, College Park, Maryland 20742 (A.M.).
2
Institute of Biotechnology (R.S., X.W., S.R.Y., S.L., R.U., I.S., J.Z., A.P.M.) and Department of Biosciences, Viikki Plant Science Centre (R.S., X.W., S.R.Y., S.L., R.U., I.S., J.Z., M.W., A.P.M.), University of Helsinki, Helsinki 00014, Finland;Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, Indiana 47907 (A.M., G.M.);Molecular Genetics, Department of Biology, Utrecht University, Utrecht 3584 CH, The Netherlands (M.G., R.H., B.S.);School of Biology, Indian Institute of Science Education and Research, Thiruvananthapuram 695016, Kerala, India (K.P.);Plant Developmental Biology, Wageningen University, Wageningen 6708 PB, The Netherlands (R.H., B.S.); andDepartment of Plant Science and Landscape Architecture, University of Maryland, College Park, Maryland 20742 (A.M.) aripekka.mahonen@helsinki.fi.

Abstract

A powerful method to study gene function is expression or overexpression in an inducible, cell type-specific system followed by observation of consequent phenotypic changes and visualization of linked reporters in the target tissue. Multiple inducible gene overexpression systems have been developed for plants, but very few of these combine plant selection markers, control of expression domains, access to multiple promoters and protein fusion reporters, chemical induction, and high-throughput cloning capabilities. Here, we introduce a MultiSite Gateway-compatible inducible system for Arabidopsis (Arabidopsis thaliana) plants that provides the capability to generate such constructs in a single cloning step. The system is based on the tightly controlled, estrogen-inducible XVE system. We demonstrate that the transformants generated with this system exhibit the expected cell type-specific expression, similar to what is observed with constitutively expressed native promoters. With this new system, cloning of inducible constructs is no longer limited to a few special cases but can be used as a standard approach when gene function is studied. In addition, we present a set of entry clones consisting of histochemical and fluorescent reporter variants designed for gene and promoter expression studies.

PMID:
26644504
PMCID:
PMC4734558
DOI:
10.1104/pp.15.01246
[Indexed for MEDLINE]
Free PMC Article

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