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Journal of Virology, May 2003, p. 6082-6086, Vol. 77, No. 10
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.10.6082-6086.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

In Situ Characterization of Cymbidium Ringspot Tombusvirus Infection-Induced Posttranscriptional Gene Silencing in Nicotiana benthamiana

Zoltán Havelda,1* Csaba Hornyik,1 Aniello Crescenzi,2 and József Burgyán1

Agricultural Biotechnology Centre, H-2101 Gödöllõ, Hungary,1 Dipartimento di Biologia, Difesa e biotecnologie Agro-Forestali, Universita degli Studi della Basilicata, Campus Macchia Romana, I-85100 Potenza, Italy2

Received 6 November 2002/ Accepted 17 February 2003

In plants, posttranscriptional gene silencing (PTGS) is an ancient and effective defense mechanism against viral infection. A number of viruses encode proteins that suppress virus-activated PTGS. The p19 protein of tombusviruses is a potent PTGS suppressor which interferes with the onset of PTGS-generated systemic signaling and is not required for viral replication or for viral movement in Nicotiana benthamiana. This unique feature of p19 suppressor allowed us to analyze the mechanism of PTGS-based host defense and its viral suppression without interfering with other viral functions. In contrast to the necrotic symptoms caused by wild-type tombusvirus, the infection of p19-defective mutant virus results in the development of a typical PTGS-associated recovery phenotype in N. benthamiana. In this report we show the effect of PTGS on the viral infection process for N. benthamiana infected with either wild-type Cymbidium Ringspot Tombusvirus (CymRSV) or a p19-defective mutant (Cym19stop). In situ analyses of different virus-derived products revealed that PTGS is not able to reduce accumulation of virus in primary infected cells regardless of the presence of p19 PTGS suppressor. We also showed that both CymRSV and Cym19stop viruses move systemically in the vasculature, with similar efficiencies. However, in contrast to the uniform accumulation of CymRSV throughout systemically infected leaves, the presence of Cym19stop virus was confined to and around the vascular bundles. These results suggest that the role of p19 is to prevent the onset of mobile signal-induced systemic PTGS ahead of the viral infection front, leading to generalized infection.

* Corresponding author. Mailing address: Agricultural Biotechnology Centre, Szent-Györgyi Albert út 4, P.O. Box 411, H-2101 Gödöllõ, Hungary. Phone: 36-28-526-100. Fax: 36-28-526-145. E-mail: havelda{at}abc.hu.

Journal of Virology, May 2003, p. 6082-6086, Vol. 77, No. 10
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.10.6082-6086.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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