AtMCP1b, a chloroplast-localised metacaspase, is induced in vascular tissue after wounding or pathogen infection
Luis Castillo-Olamendi A C , Armando Bravo-Garcìa A B C , Julio Morán B , Mario Rocha-Sosa A and Helena Porta A DA Departmento de Biología Molecular de Plantas, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Mor., México.
B Departmento de Neurociencias, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Cuernavaca, Mor., México.
C These authors contributed equally to this article.
D Corresponding author. Email: helena@ibt.unam.mx
Functional Plant Biology 34(12) 1061-1071 https://doi.org/10.1071/FP07153
Submitted: 19 June 2007 Accepted: 17 October 2007 Published: 27 November 2007
Abstract
cDNA corresponding to the Arabidopsis type I metacaspase AtMCP1b was isolated from plants infected with Pseudomonas syringae. A positive correlation between AtMCP1b expression and cell death was observed in the presence of staurosporine, a protein kinase inhibitor that induces programmed cell death. The tissue localisation of an AtMCP1b promoter-GUS fusion was observed in the vascular tissue of transgenic plants. GUS activity increased in response to an incompatible DC3000 (avrRpm1) or a compatible DC3000 P. syringae infection, or to wounding. Confocal and immunohistochemical analysis of Arabidopsis thaliana (L.) leaves showed that an AtMCP1b-GFP fusion protein was localised in the chloroplasts. Our data support a positive correlation between AtMCP1b gene expression and cell death in response to wounding or pathogenic interactions. Moreover, the localisation of AtMCP1b gene expression within vascular tissue and cells of abscission regions strongly supports a role for AtMCP1b in programmed cell dismantling events in response to environmental and developmental triggers. The AtMCP1b-GFP subcellular localisation infers a role for the plastid organelles in PCD and, thus, in responses to pathogen attack and development.
Additional keywords: chloroplast metacaspase, wounding, pathogen infection.
Acknowledgements
We thank Guadalupe Zavala and Patricia Rueda for technical support, Eugenio López-Bustos and Paul Gaytán for oligonucletoide synthesis, Jorge Yáñez for sequencing, Andrés Saralegui for confocal images and José J. Sánchez Serrano from CNB, Madrid, Spain for providing us with the Arabidopsis cell culture. We also thank Patricia León, Federico Sánchez and Miguel Angel Cevallos for helpful comments on the manuscript. This work was funded by the Dirección General de Asuntos Para el Personal Acadèmico-UNAM (IN212103).
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