APS Abstract of Presentation

In vitro transcripts of a full-length cDNA clone of Hosta virus X are infectious to Hosta and Nicotiana benthamiana plants
C. DE LA TORRE (1), D. J. Lewandowski (1)
(1) Ohio State University, Columbus, OH, USA
Phytopathology 99:S27

Hosta virus X (HVX) is an easily transmitted potexvirus that causes important economic losses to the Hosta market. No significant source of resistance to this virus is known, and information regarding the virus biology is limited. The aim of this work was to build an HVX infectious clone, which can be used both as a system to study virus-host interactions and as a tool to screen for resistance among the diverse Hosta germplasm. Hosta ‘Sum and Substance’ and Nicotiana benthamiana plants were rub-inoculated with in vitro RNA transcripts produced with T7 RNA polymerase from pHVX, a full-length cDNA clone of the Ohio HVX isolate H37. Inoculated leaves were tested by DAS-ELISA and Immunocapture RT-PCR (IC-RT-PCR) at 18 dpi. Hosta and N. benthamiana plants were positive for HVX by DAS-ELISA, with higher OD₄₀₅ values for the Hosta cultivar. IC-RT-PCR resulted in the production of an amplicon internal to the replicase open reading frame (ORF). An amplicon of expected size was also produced by RT-PCR from total RNA extracted from in vitro RNA transcript-inoculated plants using primers flanking the coat protein ORF.

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