JVI Accepts, published online ahead of print on 22 August 2007

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J. Virol. doi:10.1128/JVI.01165-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Experimental transmission of Pospiviroid populations to weed species characteristic for potato and hop fields

J. Matouek, L. Orctová, J. Ptáek, J. Patzak, P. Ddi, G. Steger, and D. Riesner^*

Department of Molecular Genetics, Biology Centre of the ASCR, v.v.i. Institute of Plant Molecular Biology, Braniovská 31, 370 05 eské Budjovice, Czech Republic; Faculty of Biological Sciences, University of South Bohemia, Braniovská 31, 37005, eské Budjovice, Czech Republic; Institute for Potato Research, Dobrovského 2366, 58001 Havlíkv Brod, Czech Republic; Hop Research Institute GmbH, Kadaská 2525, 438 46 atec, Czech Republic; Institute of Physical Biology, Heinrich-Heine-Universität Düsseldorf, Universitätsstrasse 1, D-40225 Düsseldorf, Germany

^* To whom correspondence should be addressed. Email: riesner{at}biophys.uni-duesseldorf.de.

Weed plants characteristic for potato and hop fields, respectively, have not been considered in the past as potential hosts that could transmit and lead to spreading of potato spindle tuber (PSTVd) and hop stunt (HSVd) viroids. To get insight into this problem, we biolistically inoculated those weed plants with viroid populations either as RNA or as cDNA. New potential viroid host-species, collected in central Europe, were discovered. From twelve weed species characteristic for potato fields, high viroid levels, detectable by molecular hybridization, were maintained after both, RNA and DNA transfers in Chamomilla reculita and Anthemis arvensis. Low viroid levels, detectable by RT-PCR only, were maintained after plant inoculations with cDNA in Veronica argensis and Amaranthus retroflexus. In these two species PSTVd concentrations were 10⁵ and 10³ times, respectively, lower than in tomato as estimated by real-time PCR. From 14 weeds characteristic for hop fields, high HSVd levels were detected in Galinsoga ciliata after both, RNA and DNA transfers. HSVd was found, however, not to be transmissible by seeds of this weed species. By RT-PCR traces of HSVd were detectable in HSVd-cDNA inoculated Amaranthus retroflexus. Characteristic monomeric (+)-circular and linear viroid RNAs were present in extracts from weed species propagating viroids to high levels indicating regular replication, processing and circularization of viroid RNA in these weed species. Sequence analyses of PSTVd progenies propagated in Ch. reculita and A. arvensis showed a wide spectrum of variants related to various strains, from mild to lethal variants; the sequence variants isolated from A. retroflexus and V. argensis exhibited similarity or identity to the superlethal AS1 viroid variant. All HSVd clones from G. ciliata corresponded to a HSVdg variant, which is strongly pathogenic for European hops.