title: MultiSite Gateway-Compatible Cell Type-Specific Gene-Inducible System for Plants.
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.
authors.affiliation: 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.).
authors.forename: Riccardo
authors.initials: R
authors.lastname: Siligato
authors.affiliation: 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.).
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.affiliation: 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.).
authors.forename: Shri Ram
authors.initials: SR
authors.lastname: Yadav
authors.affiliation: 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.).
authors.forename: Satu
authors.initials: S
authors.lastname: Lehesranta
authors.affiliation: 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.).
authors.forename: Guojie
authors.initials: G
authors.lastname: Ma
authors.affiliation: 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.).
authors.forename: Robertas
authors.initials: R
authors.lastname: Ursache
authors.affiliation: 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.).
authors.forename: Iris
authors.initials: I
authors.lastname: Sevilem
authors.affiliation: 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.).
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: 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.).
authors.forename: Maartje
authors.initials: M
authors.lastname: Gorte
authors.affiliation: 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.).
authors.forename: Kalika
authors.initials: K
authors.lastname: Prasad
authors.affiliation: 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.).
authors.forename: Michael
authors.identifier: http://orcid.org/0000-0002-5946-9060
authors.initials: M
authors.lastname: Wrzaczek
authors.affiliation: 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.).
authors.forename: Renze
authors.initials: R
authors.lastname: Heidstra
authors.affiliation: 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.).
authors.forename: Angus
authors.identifier: http://orcid.org/0000-0001-5649-7413
authors.initials: A
authors.lastname: Murphy
authors.affiliation: 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.).
authors.forename: Ben
authors.identifier: http://orcid.org/0000-0001-5400-9578
authors.initials: B
authors.lastname: Scheres
authors.affiliation: 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.
authors.forename: Ari Pekka
authors.identifier: http://orcid.org/0000-0001-6051-866X
authors.initials: AP
authors.lastname: Mähönen
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000235:genetics*; D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D005822:Genetic Vectors; D009928:Organ Specificity; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins
chemical_list: D011993:Recombinant Fusion Proteins