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JXB Advance Access published online on April 29, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri164
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org
Received December 21, 2004
Accepted March 16, 2005

RESEARCH PAPER

Molecular characterization and systemic induction of single-chain ribosome-inactivating proteins (RIPs) in sugar beet (Beta vulgaris) leaves

Rosario Iglesias 1, Yolanda Pérez 1, Carlos de Torre 1, J. Miguel Ferreras 1, Pilar Antolín 1, Pilar Jiménez 1, M. Ángeles Rojo 1, Enrique Méndez 2, and Tomás Girbés 1*

1 Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Valladolid, E-47005 Valladolid, Spain
2 Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CSIC), Canto Blanco, E-28049 Madrid, Spain

* To whom correspondence should be addressed.
Tomás Girbés, E-mail: girbes{at}bio.uva.es


  Abstract

Sugar beet (Beta vulgaris L.) leaves contain virus-inducible type 1 (single chain) ribosome-inactivating proteins that have been named beetins. The structural and functional characterization, the cellular location, and the potential role of beetins as antiviral agents are reported here. Beetins are formed of a single polypeptide chain with a varying degree of glycosylation and strongly inhibited in vitro protein synthesis in rabbit reticulocyte lysates (IC50=1.15 ng ml-1) and a Vicia sativa L. cell-free system (IC50=68 ng ml-1) through the single depurination of the large rRNA. Beetins trigger the multidepurination of tobacco mosaic virus (TMV) genomic RNA which underwent extensive degradation upon treatment with acid aniline. Beetins are extracellular proteins that were recovered from the apoplastic fluid. Induction of sugar beet RIPs with either H2O2 or artichoke mottled crinkle virus (AMCV) was observed in leaves distant from the site of application of such elicitors. The external application of purified beetin to sugar leaves prevented infection by AMCV which supports the preliminary hypothesis that beetins could be involved in plant systemic acquired resistance subjected to induction by phytopathogens.

Keywords: Beetin; Beta vulgaris; protein synthesis inhibitor; ribosome-inactivating protein; sugar beet.
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