Journal Article

Promoter trapping in <i>Lotus japonicus</i> reveals novel root and nodule GUS expression domains

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

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 46, issue 8, pages 1202-1212
Published in print August 2005 | ISSN: 0032-0781
Published online August 2005 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pci129
Promoter trapping in Lotus japonicus reveals novel root and nodule GUS expression domains

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  • Biochemistry
  • Molecular and Cell Biology
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Agrobacterium-based transformation was used to introduce a promoter-less glucuronidase uidA gene (β-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 ATPase (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; ED, GUS expression domain; GUS, β-glucuronidase; LRP, lateral root primordia; NP, nodule primordia; TAC, transformation-competent artificial chromosome; Wpi, weeks post inoculation

Journal Article.  7541 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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