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Plant and Cell Physiology Advance Access published online on May 16, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci129
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received February 23, 2005
Accepted May 12, 2005

Regular Paper

Promoter Trapping in Lotus japonicus Reveals Novel Root and Nodule GUS Expression Domains

Diana Mihaela Buzas 1*, Dasharath Lohar 2, Shusei Sato 3, Yasukazu Nakamura 3, Satoshi Tabata 3, Claudia Estelle Vickers 4, Jiri Stiller 1, and Peter Michael Gresshoff 1

1 ARC Centre of Excellence for Integrative Legume Research, The University of Queensland, 4072 Australia
2 Department of Plant Biology, 250 Bio Sciences Center, 1445 Gortner Avenue, Saint Paul, MN 55108-1095
3 Kazusa DNA Research Institute, 2-6-7 Kazusa-Kamatari, Kisarazu, Chiba, 292-0812, Japan
4 ARC Centre of Excellence for Integrative Legume Research, The University of Queensland, 4072 Australia; Department of Biological Sciences, University of Essex, Wivenhoe Park, Colchester CO4 3SQ

* To whom correspondence should be addressed.
Diana Mihaela Buzas, E-mail: d.buzas{at}uq.edu.au


  Abstract

Agrobacterium-based transformation was used to introduce a promoter-less glucuronidase uidA gene ({beta}-glucuronidase; GUS) into Lotus japonicus. Transgenic plants were screened for GUS activation at different stages after inoculation with its symbiont, Mesorhizobium loti. Functional GUS fusion frequencies ranged from about 2 to 5% of the total number of transgenic lines. These lines provide excellent histological markers for tissue ontogeny analysis. Some of the activations generated GUS expression patterns that correspond to well-known tissue types, such as lateral root and nodule primordia, root tips and developing nodules (line CHEETAH). Others generated GUS activation associated with predictable but previously unknown i) tissue types, such as the vascular bundle of the nodule (line VASCO) or ii) expression domains, such as pericycle, nodule primordia, nodule and flower connective/vascular tissue (line FATA MORGANA) or inner root cortex cells in the vicinity of a curled root hair, nodule primordia and nodule cortex (line TIMPA). Putative members of two gene superfamilies, EH (Esp homolog) and AAA ATP-ase (ATPase associated with various cellular activities), were located next to the CHEETAH and VASCO insertions, respectively and a nodulin gene, LjENOD40-2, was located next to the FATA MORGANA insertion. We utilized promoter GUS fusions to investigate the genetic regulation of LjENOD40-2 and FATA MORGANA GUS. The LjENOD40-2 promoter defined a novel expression domain and the FATA MORGANA nodule expression was reiterated by the 2 kb sequence upstream of the T-DNA insertion.

Keywords: ENOD40; GUS expression domain; Lotus japonicus; nodule development.
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