J. Mertelik1*, K. Kloudova1, G. Cervena2, J. Necekalova2, H. Mikulkova2, Z. Levkanicova2, P. Dedic3 and J. Ptacek3
1 Silva Tarouca Research Institute for Landscape and Ornamental Gardening, Publ. Res Inst., Pruhonice, Czech Republic
2 State phytosanitary Administration, Division of diagnostic, Olomouc, Czech Republic
3 Potato Research Instutute, Havlickuv Brod, Czech Republic
Accepted: 06 Apr 2009
Following findings of Potato spindle tuber viroid (PSTVd) in solanaceous ornamental plants in The Netherlands (Verhoeven et al., 2007) and other European countries the survey of PSTVd was carried out also in the Czech Republic.
Samples were collected from gardening establishments and private gardens. A total of 156 samples (52 of S. jasminoides, 65of Brugmansia spp., 9 of S. muricatum and 30 of Petunia x hybrida) were taken from 65 localities. Samples consisted of ten randomly taken leaves per lot or in case of Brugmansia spp. and Solanum muricatum of pooled samples of five to ten leaves taken from individual plants.
RNA was extracted from leaf tissue using either RNeasy Plant Mini Kit (Qiagen) or Tri Reagent (Sigma). The samples were tested by real-time RT-PCR using SYBR Green with PSTVd primers (Boonham et al., 2004 and Matoušek et al., 2007) and RT-PCR using PSTVd primers (Levy, 1994) that yielded approximately 360 bp long amplicons. The PCR products were sequenced and used for identification. All the sequences obtained were identical to the isolate of PSTVd GenBank Accession No. EF459700.
S. jasminoides tested positively for PSTVd at most of the sampled localities due to repeated vegetative propagation of infected plants and their subsequent spread throughout the country. Petunia x hybrida tested positively for PSTVd only at 2 and Brugmansia spp. and S. muricatum at 1 locality. To our knowledge, this is the first report of PSTVd in Brugmansia spp., Solanum jasminoides, Solanum muricatum and Petunia x hybrida in the CzechRepublic.
Boonham N, Gonzáles Pérez L, Mendez MS, Lilia Peralta E, Blockly A, Walsh K, Barker I, Mumford RA, 2004. Development of a real-time RT-PCR assay for the detection of Potato spindle tuber viroid. Journal of Virological Methods 116, 139-146.
Levy, L., Lee, I.-M., and Hadidi, A. 1994. Simple and rapid preparation of infected plant tissue extracts for PCR amplification of virus, viroid, and MLO nucleic acids. Journal of Virological Methods 49, 295-304.
Matoušek, J., Orctová, L., Ptáček, J., Patzak, J., Dědič, P., Steger, G., Riesner, D. 2007. Experimental transmission of Pospiviroid populations to weed species characteristic for potato and hopfields. Journal of Virology 81, 11891-11899.
Verhoeven JThJ, Jansen CCC, Roenhorst, JW, 2007.First report of pospiviroids infecting ornamentals in the Netherlands: Citrus exocortis viroid in Verbena sp., Potato spindle tuber viroid in Brugmansia suaveolens and Solanum jasminoides,and Tomato apical stunt viroid in Cestrum sp. New Disease Reports [http://www.ndrs.org.uk/july2007/2007-13.asp] Volume 15.
©2009 The Authors