Open AccessFirst report of Columnea latent viroid (CLVd)in Gloxinia gymnostoma, G. nematanthodes and G. purpurascens in a botanical garden in Denmark
Author(s) -
Nielsen S.L.,
Nicolaisen M.
Publication year - 2010
Publication title -
new disease reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.184
H-Index - 4
ISSN - 2044-0588
DOI - 10.5197/j.2044-0588.2010.022.004
Subject(s) - genbank , biology , ornamental plant , accession number (library science) , viroid , botany , virology , genetics , plant virus , gene , virus
RNA was extracted from 100 mg fresh leaf material using the RNeasy Plant Mini Kit (Qiagen). RT-PCR was carried out using the Vid primers of Verhoeven et al. (2004) and the pCLVd4 and pCLVdR4 primers of Spieker (1996) with the OneStep RT-PCR Kit (Qiagen). An isolate of CLVd (9389007481), kindly provided by J.Th.J. Verhoeven, was used as positive control. A product of the expected size (c. 350 bp) was obtained from all three Gloxinia species and was subsequently sequenced. A BLAST search in the NCBI GenBank confirmed all three samples to be CLVd. However, as the sequences indicated presence of more than one genotype in the three Gloxinia species, the PCR products were cloned using the Stratagene TM