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Development of an RT-PCR/RFLP assay to detect Hosta virus X in hosta (Hosta spp.)

Development of an RT-PCR/RFLP assay to detect Hosta virus X in hosta (Hosta spp.)

Journal of Horticultural Science and Biotechnology 86(1): 50-54

ISSN/ISBN: 1462-0316

DOI: 10.1080/14620316.2011.11512724

Hosta virus X (HVX) is the most damaging virus of hosta (Hosta spp.). By taking advantage of the conserved coat protein (CP) sequences of characterised isolates of HVX, a reliable and sensitive diagnostic assay, based on RT-PCR, was developed. The conserved, single Hind II recognition site within the CP sequences was also exploited for restriction fragment length polymorphism (RFLP)-based verification of the amplicons. The reliability of the assay was demonstrated by successful amplification of 30 HVX isolates from 20 cultivars of hosta. The effectiveness of the assay was demonstrated by detecting several distinct isolates of HVX in a mechanically-inoculated hosta cultivar containing a low virus titre. The assay was capable of detecting HVX in composite samples consisting of a single HVX-infected leaf disc, approx. 1 cm in diameter, combined with up to 200 virus-free leaf discs of similar size, which demonstrated its potential for large-scale screening. The sensitivity of the assay was not affected by genetic variation in the hosta cultivars or in the virus isolates. We conclude that RT-PCR/RFLP is reliable, sensitive, and efficient for large-scale screening of hosta for HVX infection.

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Accession: 066241967

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