Published online 18 January 2006
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Molecular characterization of geminivirus-derived small RNAs in different plant species
1Institute of Botany, University of Basel Schönbeinstrasse 6, 4056 Basel, Switzerland 2Friedrich Miescher Institute for Biomedical Research Maulbeerstrasse 66, 4058 Basel, Switzerland 3Institute of Plant Sciences, ETH-Zurich, LFW E17, Universitätstrasse 2 8092 Zürich, Switzerland
*To whom correspondence should be addressed. Tel: +4161 2672977; Fax: +4161 2673504; Email: Mikhail.Pooggin{at}unibas.ch
Received December 2, 2005. Revised December 23, 2005. Accepted January 3, 2006.
DNA geminiviruses are thought to be targets of RNA silencing. Here, we characterize small interfering (si) RNAs—the hallmarks of silencing—associated with Cabbage leaf curl begomovirus in Arabidopsis and African cassava mosaic begomovirus in Nicotiana benthamiana and cassava. We detected 21, 22 and 24 nt siRNAs of both polarities, derived from both the coding and the intergenic regions of these geminiviruses. Genetic evidence showed that all the 24 nt and a substantial fraction of the 22 nt viral siRNAs are generated by the dicer-like proteins DCL3 and DCL2, respectively. The viral siRNAs were 5' end phosphorylated, as shown by phosphatase treatments, and methylated at the 3'-nucleotide, as shown by HEN1 miRNA methylase-dependent resistance to ß-elimination. Similar modifications were found in all types of endogenous and transgene-derived siRNAs tested, but not in a major fraction of siRNAs from a cytoplasmic RNA tobamovirus. We conclude that several distinct silencing pathways are involved in DNA virus-plant interactions.
Correspondence may also be addressed to Thomas Hohn. Tel: +4161 6977266; Fax: +4161 6973976; Email: hohn{at}fmi.ch
Present addresses: Imran Amin, National Institute for Biotechnology and Genetic Engineering, Jhang Road, Faisalabad, Pakistan
Peng Zhang, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, 300 Fenglin Road, 200032 Shanghai, China
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