TRANSPLANTA collection of Arabidopsis lines: a resource for functional analysis of transcription factors based on their conditional overexpression
Transcription factors (TFs) are key regulators of gene expression in all organisms. In eukaryotes, TFs are often represented by functionally redundant members of large gene families. Overexpression might prove a means to unveil the biological functions of redundant TFs; however, constitutive overexp...
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Published in | The Plant journal : for cell and molecular biology Vol. 77; no. 6; pp. 944 - 953 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
England
Blackwell Publishing Ltd
01.03.2014
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Subjects | |
Online Access | Get full text |
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Abstract | Transcription factors (TFs) are key regulators of gene expression in all organisms. In eukaryotes, TFs are often represented by functionally redundant members of large gene families. Overexpression might prove a means to unveil the biological functions of redundant TFs; however, constitutive overexpression of TFs frequently causes severe developmental defects, preventing their functional characterization. Conditional overexpression strategies help to overcome this problem. Here, we report on the TRANSPLANTA collection of Arabidopsis lines, each expressing one of 949 TFs under the control of a β–estradiol‐inducible promoter. Thus far, 1636 independent homozygous lines, representing an average of 2.6 lines for every TF, have been produced for the inducible expression of 634 TFs. Along with a GUS‐GFP reporter, randomly selected TRANSPLANTA lines were tested and confirmed for conditional transgene expression upon β–estradiol treatment. As a proof of concept for the exploitation of this resource, β–estradiol‐induced proliferation of root hairs, dark‐induced senescence, anthocyanin accumulation and dwarfism were observed in lines conditionally expressing full‐length cDNAs encoding RHD6, WRKY22, MYB123/TT2 and MYB26, respectively, in agreement with previously reported phenotypes conferred by these TFs. Further screening performed with other TRANSPLANTA lines allowed the identification of TFs involved in different plant biological processes, illustrating that the collection is a powerful resource for the functional characterization of TFs. For instance, ANAC058 and a TINY/AP2 TF were identified as modulators of ABA‐mediated germination potential, and RAP2.10/DEAR4 was identified as a regulator of cell death in the hypocotyl–root transition zone. Seeds of TRANSPLANTA lines have been deposited at the Nottingham Arabidopsis Stock Centre for further distribution. |
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AbstractList | Summary Transcription factors (TFs) are key regulators of gene expression in all organisms. In eukaryotes, TFs are often represented by functionally redundant members of large gene families. Overexpression might prove a means to unveil the biological functions of redundant TFs; however, constitutive overexpression of TFs frequently causes severe developmental defects, preventing their functional characterization. Conditional overexpression strategies help to overcome this problem. Here, we report on the TRANSPLANTA collection of Arabidopsis lines, each expressing one of 949 TFs under the control of a [beta]-estradiol-inducible promoter. Thus far, 1636 independent homozygous lines, representing an average of 2.6 lines for every TF, have been produced for the inducible expression of 634 TFs. Along with a GUS-GFP reporter, randomly selected TRANSPLANTA lines were tested and confirmed for conditional transgene expression upon [beta]-estradiol treatment. As a proof of concept for the exploitation of this resource, [beta]-estradiol-induced proliferation of root hairs, dark-induced senescence, anthocyanin accumulation and dwarfism were observed in lines conditionally expressing full-length cDNAs encoding RHD6, WRKY22, MYB123/TT2 and MYB26, respectively, in agreement with previously reported phenotypes conferred by these TFs. Further screening performed with other TRANSPLANTA lines allowed the identification of TFs involved in different plant biological processes, illustrating that the collection is a powerful resource for the functional characterization of TFs. For instance, ANAC058 and a TINY/AP2 TF were identified as modulators of ABA-mediated germination potential, and RAP2.10/DEAR4 was identified as a regulator of cell death in the hypocotyl-root transition zone. Seeds of TRANSPLANTA lines have been deposited at the Nottingham Arabidopsis Stock Centre for further distribution. [PUBLICATION ABSTRACT] Transcription factors (TFs) are key regulators of gene expression in all organisms. In eukaryotes, TFs are often represented by functionally redundant members of large gene families. Overexpression might prove a means to unveil the biological functions of redundant TFs; however, constitutive overexpression of TFs frequently causes severe developmental defects, preventing their functional characterization. Conditional overexpression strategies help to overcome this problem. Here, we report on the TRANSPLANTA collection of Arabidopsis lines, each expressing one of 949 TFs under the control of a β-estradiol-inducible promoter. Thus far, 1636 independent homozygous lines, representing an average of 2.6 lines for every TF, have been produced for the inducible expression of 634 TFs. Along with a GUS-GFP reporter, randomly selected TRANSPLANTA lines were tested and confirmed for conditional transgene expression upon β-estradiol treatment. As a proof of concept for the exploitation of this resource, β-estradiol-induced proliferation of root hairs, dark-induced senescence, anthocyanin accumulation and dwarfism were observed in lines conditionally expressing full-length cDNAs encoding RHD6, WRKY22, MYB123/TT2 and MYB26, respectively, in agreement with previously reported phenotypes conferred by these TFs. Further screening performed with other TRANSPLANTA lines allowed the identification of TFs involved in different plant biological processes, illustrating that the collection is a powerful resource for the functional characterization of TFs. For instance, ANAC058 and a TINY/AP2 TF were identified as modulators of ABA-mediated germination potential, and RAP2.10/DEAR4 was identified as a regulator of cell death in the hypocotyl-root transition zone. Seeds of TRANSPLANTA lines have been deposited at the Nottingham Arabidopsis Stock Centre for further distribution. Summary Transcription factors (TFs) are key regulators of gene expression in all organisms. In eukaryotes, TFs are often represented by functionally redundant members of large gene families. Overexpression might prove a means to unveil the biological functions of redundant TFs; however, constitutive overexpression of TFs frequently causes severe developmental defects, preventing their functional characterization. Conditional overexpression strategies help to overcome this problem. Here, we report on the TRANSPLANTA collection of Arabidopsis lines, each expressing one of 949 TFs under the control of a β–estradiol‐inducible promoter. Thus far, 1636 independent homozygous lines, representing an average of 2.6 lines for every TF, have been produced for the inducible expression of 634 TFs. Along with a GUS‐GFP reporter, randomly selected TRANSPLANTA lines were tested and confirmed for conditional transgene expression upon β–estradiol treatment. As a proof of concept for the exploitation of this resource, β–estradiol‐induced proliferation of root hairs, dark‐induced senescence, anthocyanin accumulation and dwarfism were observed in lines conditionally expressing full‐length cDNAs encoding RHD6, WRKY22, MYB123/TT2 and MYB26, respectively, in agreement with previously reported phenotypes conferred by these TFs. Further screening performed with other TRANSPLANTA lines allowed the identification of TFs involved in different plant biological processes, illustrating that the collection is a powerful resource for the functional characterization of TFs. For instance, ANAC058 and a TINY/AP2 TF were identified as modulators of ABA‐mediated germination potential, and RAP2.10/DEAR4 was identified as a regulator of cell death in the hypocotyl–root transition zone. Seeds of TRANSPLANTA lines have been deposited at the Nottingham Arabidopsis Stock Centre for further distribution. |
Author | León, José Coego, Alberto Paz‐Ares, Javier Jarillo, José A Koncz, Csaba Ruíz, Sandra Castillejo, Pablo Brizuela, Esther del Pozo, Juan C Piñeiro, Manuel |
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BackLink | https://www.ncbi.nlm.nih.gov/pubmed/24456507$$D View this record in MEDLINE/PubMed |
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Contributor | Rapp, Ana Coego, Alberto Paz-Ares, Javier López, Leticia del Pozo, Juan Carlos Resa, Carlos Perea González, Mar Abraham, Edit Ballesteros, Isabel del Olmo, Ivan Ferrándiz, Cristina Rajulu, Charukesi Madueño, Francisco del Llano, Barbara Carrió, Angela Felipo, Amelia Leyva, Antonio Quintero, Luis A Castresana, Carmen Koncz, Zsuzsa Alabadí, David Olsson, Sanna Boter, Marta Pardo, José Manuel Alonso, Carlos Piñeiro, Manuel Solano, Roberto Rojo, Enrique Romero, Luis Carlos Alvarez-Mahecha, Juan C García, Alvaro Zsigmond, Laura Castro, Ruth Brizuela, Esther Cubero, Beatriz Gómez, Marcela Andrés, Nuria Esteban, Alicia Gotor, Cecilia Papdi, Csaba Cséplő, Ágnes Peñarrubia, Lola Botella, Miguel A Fenoll, Carmen Berbel, Ana Gutierrez, Crisanto Manzano, Concepcion Lopez, Cristian Carrasco Jover-Gil, Sara Carbajosa, Jesús Vicente Minguet, Eugenio G Sotillo, Berta de Luca, Anna Navarro, Sara Székely, Gyöngyi Cubas, Pilar Fernández, Guillermo M Arocas, Loli Vera, Pablo Narro, Laura Delgadillo, Ma Otilia Diéz-Díaz, Mónica Luque, Nazaret Nicolas, Michael Codes, Cr |
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Copyright | 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd. Copyright © 2014 John Wiley & Sons Ltd and the Society for Experimental Biology |
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Keywords | Arabidopsis thaliana transgenic lines collection TRANSPLANTA consortium transcription factors functional genomics conditional overexpression functional screening |
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Snippet | Transcription factors (TFs) are key regulators of gene expression in all organisms. In eukaryotes, TFs are often represented by functionally redundant members... Summary Transcription factors (TFs) are key regulators of gene expression in all organisms. In eukaryotes, TFs are often represented by functionally redundant... Summary Transcription factors (TFs) are key regulators of gene expression in all organisms. In eukaryotes, TFs are often represented by functionally redundant... |
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SubjectTerms | Abscisic Acid - metabolism anthocyanins Arabidopsis Arabidopsis - genetics Arabidopsis - growth & development Arabidopsis - metabolism Arabidopsis Proteins - genetics Arabidopsis Proteins - metabolism Arabidopsis thaliana cell death complementary DNA conditional overexpression DNA, Complementary - genetics Estradiol - metabolism eukaryotic cells functional genomics functional screening Gene Expression gene overexpression Genes, Reporter Genetic Vectors Germination Phenotype Plant biology Plant Growth Regulators - metabolism Plant Roots - genetics Plant Roots - metabolism Plants, Genetically Modified Promoter Regions, Genetic - genetics regulator genes root hairs screening Seedlings - genetics Seedlings - metabolism seeds Seeds - genetics Seeds - metabolism senescence Transcription factors Transcription Factors - genetics Transcription Factors - metabolism Transcriptional Activation - drug effects Transgenes transgenic lines collection TRANSPLANTA consortium |
Title | TRANSPLANTA collection of Arabidopsis lines: a resource for functional analysis of transcription factors based on their conditional overexpression |
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