Complete nucleotide sequence and molecular probing of potato virus S genome
Matousek, J.(Czech Academy of Sciences, Ceske Budejovice (Czech Republic). Institute of Plant Molecular Biology | Schubert, J.Federal Center for Breeding Research on Cultivated Plants, Aschersleben (Germany) | Ptacek, J.Institute for Potato Research, Havlickuv Brod (Czech Republic) | Kozlova,P.Czech Academy of Sciences, Ceske Budejovice (Czech Republic). Institute of Plant Molecular Biology | Dedic, P.Federal Center for Breeding Research on Cultivated Plants, Aschersleben (Germany))
Complete genomes of three isolates of Potato virus S (PVS) were cloned and sequenced. The PVS ORF-1 was characterized for the first time. It encodes a putative replication protein (RPT) that shares the highest homology (about 52%) with that of Blueberry scorch virus (BIScV). ORF-1 motifs, characteristic for carlaviruses were found for methyl trans ferase (MTR), helicase (HEL) and RNA-dependent RNA polymerase (RdRp). The complete sequence of PVS genome enabled to develop an immunocapture RT-PCR probing of the PVS genome. Using this system, the sequence variability of 11 genome zones was examined for 34 PVS isolates including 15 PVS-CS variants that caused a systemic infection in Chenopodium quinoa. A broad variability between PVS isolates and diverse sequence variants was found. cDNA fragments covering the coat protein (CP) leader and CP-coding region (approx. 420 bp) were pooled for PVS-0 and Chenopodium-systemic PVS isolates (PVS-CS) and corresponding cDNA libraries were screened for sequence variants. Both cDNA pools differred mainly in the 5-end of the CP gene. Methionine at the position 17 in combination with serine at the position 34 were frequently associated with the CS character of PVS. In general, hydrophobic and polar amino acids were characteristic for the positions 17 and 34, respectively in PVS-CS isolates. Genome probing and evolutionary distances suggested that the PVS-CS isolates analyzed were close to the ordinary European isolates of ordinary strain of PVS (PVS-O) but distant to the original Andean strain of PVS (PVS-A).
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