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Plant and Cell Physiology Advance Access first published online on August 30, 2006
This version published online on September 5, 2006
Plant and Cell Physiology, doi:10.1093/pcp/pcj102
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© The Author 2006. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org
Received June 27, 2006
Accepted August 3, 2006

Regular Paper

Robotized thermal and chlorophyll-fluorescence imaging of pepper mild mottle virus infection in Nicotiana benthamiana

Laury Chaerle 1, Mónica Pineda 2, Remedios Romero-Aranda 3, Dominique Van Der Straeten 4 *, and Matilde Barón 5 *

1 Unit Plant Hormone Signaling and Bioimaging, Ghent University, K. L. Ledeganckstraat 35, B-9000 Gent (Belgium).; These authors have equally contributed to this work and are placed in alphabetical order
2 Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín. C/ Profesor Albareda, n° 1. C.P. 18008. Granada (Spain).; These authors have equally contributed to this work and are placed in alphabetical order
3 Department of Plant Breeding, Estación Experimental La Mayora. 29750 Algarrobo-Costa, Málaga (Spain).
4 Unit Plant Hormone Signaling and Bioimaging, Ghent University, K. L. Ledeganckstraat 35, B-9000 Gent (Belgium).
5 Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín. C/ Profesor Albareda, n° 1. C.P. 18008. Granada (Spain).

* To whom correspondence should be addressed.
Dominique Van Der Straeten, E-mail: dominique.vanderstraeten{at}ugent.be
Matilde Barón, E-mail: mbaron{at}eez.csic.es


  Abstract

After infecting a susceptible host, pathogens spread throughout the plant. Depending on pathogen type and strain, the severity of symptoms greatly varies. In the case of pepper mild mottle virus (PMMoV) infection in Nicotiana benthamiana, newly developing leaves display visual symptoms (symptomatic leaves). In this study two PMMoV strains were used, the Spanish strain (PMMoV-S) being more virulent than the Italian one (PMMoV-I). Plants infected with PMMoV-I could recover from the virus-induced symptoms. Leaves that were fully developed at the start of PMMoV infection remained symptomless. In these asymptomatic leaves, an increase in temperature, initiating from the tissue adjacent to the main veins, was observed 7 days before the chlorophyll fluorescence pattern changed. Virus immunolocalization on tissue prints matched well with the concomitant pattern of chlorophyll fluorescence increase. The temperature increase, associated with the veins, was shown to be related to stomatal closure. Upon PMMoV-I infection, appearance of thermal and chlorophyll fluorescence symptoms as well as virus accumulation were delayed by 3 days compared to PMMoV-S induced symptoms. Temperature increase of whole symptomatic leaves was also correlated to a decrease in stomatal aperture. In contrast to the persistent increase in symptomatic leaf temperature observed during PMMoV-S infection, the temperature of symptomatic leaves of PMMoV-I infected plants decreased gradually during recovery.

We propose that the earliest temperature increase is caused by a systemic plant response to the virus infection, involving the control of water loss. In conclusion, thermography has a potential as an early reporter of an ongoing compatible infection process.

Keywords: Biotic stress; Infrared gas analysis; Nicotiana benthamiana; Pepper mild mottle virus; Stomatal conductance; Thermography.
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