TRANSPLANTA collection of Arabidopsis lines: a resource for functional analysis of transcription factors based on their conditional overexpression

• Published in: The Plant journal : for cell and molecular biology Vol. 77; no. 6; pp. 944 - 953
• Main Authors: Coego, Alberto, Brizuela, Esther, Castillejo, Pablo, Ruíz, Sandra, Koncz, Csaba, del Pozo, Juan C, Piñeiro, Manuel, Jarillo, José A, Paz‐Ares, Javier, León, José
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.03.2014
• Subjects: 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; Arabidopsis thaliana; transgenic lines collection; TRANSPLANTA consortium; transcription factors; functional genomics; conditional overexpression; functional screening
• ISSN: 0960-7412; 1365-313X
• DOI: 10.1111/tpj.12443

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.