The authors have no conflict of interest to declare regarding the publication of this paper.
A GAL4-based targeted activation tagging system in Arabidopsis thaliana
Article first published online: 23 NOV 2012
© 2012 The Authors The Plant Journal © 2012 Blackwell Publishing Ltd
The Plant Journal
Volume 73, Issue 3, pages 357–367, February 2013
How to Cite
Waki, T., Miyashima, S., Nakanishi, M., Ikeda, Y., Hashimoto, T. and Nakajima, K. (2013), A GAL4-based targeted activation tagging system in Arabidopsis thaliana. The Plant Journal, 73: 357–367. doi: 10.1111/tpj.12049
Accession number: GenBank accession number for URP1 locus, JX308239.
- Issue published online: 28 JAN 2013
- Article first published online: 23 NOV 2012
- Accepted manuscript online: 12 OCT 2012 05:05AM EST
- Manuscript Accepted: 9 OCT 2012
- Manuscript Revised: 20 SEP 2012
- Manuscript Received: 27 JUL 2012
- Japan Society for Promotion of Science (JSPS). Grant Numbers: 15770146, 18510171, 20061022 and 21027025
- Novartis Foundation
- The Sumitomo Foundation. Grant Number: 060507
- activation tagging;
- Arabidopsis thaliana ;
- pattern formation;
- transcription factor;
- technical advance
Activation tagging is a powerful tool for discovering novel genes that are not easily identified by loss-of-function (lof) screening due to genetic redundancy or lethality. Although the current activation tagging system, which involves a viral enhancer sequence, has been used for a decade, alternative methods that allow organ- or tissue-specific activation are required to identify genes whose strong activation leads to loss of fertility or viability. Here, we established a GAL4/UAS activation-tagging system in Arabidopsis thaliana. Host plants that express a synthetic transcription activator GAL4:VP16 (GV) in an organ- or tissue-specific manner were transformed with a T-DNA harboring tandem copies of UAS, a GAL4-binding sequence. Using a post-embryonic and root-specific GV-expressing line as the host plant, we isolated several dominant mutants with abnormal root tissue patterns, designated as uas-tagged root patterning (urp) mutants, and identified their causal genes. Notably, most URP genes encoded putative transcription factors, indicating that the GAL4/UAS activation tagging system effectively identifies genes with regulatory functions. lof phenotypes of most URP genes were either local patterning defects or visible only if homologous genes were disrupted simultaneously or independently. Systemic overexpression of some URP genes resulted in seedling lethality. These results indicate that GAL4/UAS activation tagging is a powerful method for identifying genes with biological functions that are not readily identified by conventional screening methods.