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Journal of Virology, January 2005, p. 450-457, Vol. 79, No. 1
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.1.450-457.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Defective Interfering RNA Hinders the Activity of a Tombusvirus-Encoded Posttranscriptional Gene Silencing Suppressor

Zoltán Havelda,*{dagger} Csaba Hornyik,{dagger} Anna Válóczi, and József Burgyán

Agricultural Biotechnology Center, Institute of Plant Biology, Gödöllo, Hungary

Received 20 May 2004/ Accepted 13 August 2004

Defective interfering (DI) RNAs are subviral replicons originating from the viral genome and are associated with many plant RNA viruses and nearly all animal RNA viruses. The presence of DI RNAs in tombusvirus-infected plants reduces the accumulation of helper virus RNA and results in the development of attenuated symptoms similar to those caused by tombusviruses defective in p19, the posttranscriptional gene silencing (PTGS) suppressor. In situ analysis of infected plants containing DI RNAs revealed that the extent of virus infection was spatially restricted as was found for p19-defective tombusvirus. Previously, p19 was shown to suppress PTGS by sequestering the small interfering RNAs (siRNAs), which act as the specificity determinant for PTGS. Our results demonstrate that DI RNAs dramatically elevate the level of virus-specific siRNAs in viral infections, resulting in the saturation of p19 and the accumulation of unbound siRNAs. Moreover, we showed that, at low temperature, where PTGS is inhibited, DI RNAs are not able to efficiently interfere with virus accumulation and protect the plants. These data show that the activation of PTGS plays a pivotal role in DI RNA-mediated interference. Our data also support a role for 21-nucleotide siRNAs in PTGS signaling.

* Corresponding author. Mailing address: Agricultural Biotechnology Center, Institute of Plant Biology, P.O. Box 411, 2100 Gödöllo, Hungary. Phone: 36 28 526100. Fax: 36 28 526145. E-mail: havelda{at}abc.hu.

{dagger} Z.H. and C.H. contributed equally to this work.

Journal of Virology, January 2005, p. 450-457, Vol. 79, No. 1
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.1.450-457.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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