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JXB Advance Access published online on November 5, 2009
Journal of Experimental Botany, doi:10.1093/jxb/erp305
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© 2009 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

RESEARCH PAPER

Ectopic expression of PtaRHE1, encoding a poplar RING-H2 protein with E3 ligase activity, alters plant development and induces defence-related responses

Johnny Mukoko Bopopi1 *, Olivier M. Vandeputte1 *, Kristiina Himanen2, Adeline Mol1, Quentin Vaessen1, Mondher El Jaziri1 and Marie Baucher1,{dagger}

1Laboratoire de Biotechnologie Végétale, Université Libre de Bruxelles, rue Adrienne Bolland 8, 6041 Gosselies, Belgium
2Department of Plant Systems Biology, Flanders Institute for Biotechnology, Technologiepark 927, 9052 Gent, Belgium

{dagger} To whom correspondence should be addressed. E-mail: mbaucher{at}ulb.ac.be

RING (really interesting new gene)-H2 domain-containing proteins are widely represented in plants and play important roles in the regulation of many developmental processes as well as in plant–environment interactions. In the present report, experiments were performed to unravel the role of the poplar gene PtaRHE1, coding for a RING-H2 protein. In vitro ubiquitination assays indicate a functional E3 ligase activity for PtaRHE1 with the specific E2 ubiquitin-conjugating enzyme UbcH5a. The overexpression of PtaRHE1 in tobacco resulted in a pleiotropic phenotype characterized by a curling of the leaves, the formation of necrotic lesions on leaf blades, growth retardation, and a delay in floral transition. The plant gene expression response to PtaRHE1 overexpression provided evidence for the up-regulation of defence- and/or programmed cell death-related genes. Moreover, genes coding for WRKY transcription factors as well as for mitogen-activated protein kinases, such as wound-induced protein kinase (WIPK), were also found to be induced in the transgenic lines as compared with the wild type. In addition, histochemical β-glucuronidase staining showed that the PtaRHE1 promoter is induced by plant pathogens and by elicitors such as salicylic acid and cellulase. Taken together, these results suggest that the E3 ligase PtaRHE1 plays a role in the ubiquitination-mediated regulation of defence response, possibly by acting upstream of WIPK and/or in the activation of WRKY factors.

Key words: Defence response, E3 ligase, Nicotiana tabacum, Populus tremulaxP. alba, RING-H2

* These authors contributed equally to this work.

Received 23 July 2009; Revised 21 September 2009 Accepted 23 September 2009


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