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Plant Physiology 134:18-27 (2004)
© 2004 American Society of Plant Biologists

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BREAKTHROUGH TECHNOLOGIES

Gene Trapping with Firefly Luciferase in Arabidopsis. Tagging of Stress-Responsive Genes1,[w]

Martha C. Alvarado, Laura M. Zsigmond, Izabella Kovács, Ágnes Cséplö, Csaba Koncz and László M. Szabados*

Institute of Plant Biology, Biological Research Center, Temesvári krt. 62, 6726-Szeged, Hungary (M.C.A., L.Z., I.K., A.C., L.M.S.); and Max-Planck-Institut für Züchtungsforschung, Carl-von-Linné-Weg 10, D-50829-Köln, Germany (C.K.)

To monitor the expression of T-DNA-tagged plant genes in vivo, a collection of 20,261 transgenic lines of Arabidopsis (Columbia-0) were generated with the promoter trap vector pTluc, which carries a promoterless firefly luc (luciferase) reporter gene linked to the right T-DNA border. By detection of bioluminescence in 3-week-old seedlings, 753 lines were identified showing constitutive, organ-specific, and stress-responsive luciferase expression patterns. To facilitate the identification of well-defined luciferase expression patterns, a pooled seed stock was established. Several lines showed sugar, salt, and abscisic acid (ABA)-inducible luciferase activity. Segregation analysis of 215 promoter trap lines indicated that about 50% of plants contained single insertions, whereas 40% carried two and 10% carried three or more T-DNA tags. Sequencing the T-DNA insert junctions isolated from 17 luciferase-expressing lines identified T-DNA tags in 5'- and 3'-transcribed domains and translational gene fusions generated by T-DNA insertions in exons and introns of Arabidopsis genes. Tissue specific expression of eight wild-type Arabidopsis genes was confirmed to be similar to the luminescence patterns observed in the corresponding luciferase-tagged lines. Here, we describe the characterization of a transcriptional luc reporter gene fusion with the WBC-type ABC transporter gene At1g17840. Expression of wild-type and luciferase-tagged At1g17840 alleles revealed similar induction by salt, glucose, and ABA treatments and gibberellin-mediated down-regulation of ABA-induced expression. These results illustrate that luciferase gene traps are well suited for monitoring the expression of stress-responsive Arabidopsis genes in vivo.

http://www.plantphysiol.org/cgi/doi/10.1104/pp.103.027151.

1 This research was supported by the European Union Fifth Framework Program (Growth Vigour and Development, no. QLK5-CT-2001-01871), by Országos Tudományos Kutatási Alap (grant no. T038375), by Oktatási Miniszténum Biotechnologia2001 (grant no. BIO-00118/2001), and by the Hungarian-German Cooperation program Tudományos és Technológiai Alapítvány (grant no. D-2/01).

[w] The online version of this article contains Web-only data.

* Corresponding author; e-mail szabados{at}nucleus.szbk.uszeged.hu; fax 36-62-433434.

Received May 21, 2003; returned for revision July 18, 2003; accepted September 11, 2003.




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