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Journal of General Virology (2000), 81, 1115-1119.
© 2000 Society for General Microbiology

Plant

Aphid transmission studies using helper component proteins of Potato virus Y expressed from a vector derived from Potato virus X

Takahide Sasayab,1, Lesley Torrance1, Graham Cowan1 and Angelika Ziegler1

Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK1

Author for correspondence: Lesley Torrance. Fax +44 1382 562426. e-mail ltorra{at}scri.sari.ac.uk

The genes encoding the helper component (HC) proteins of two strains of Potato virus Y (PVY) were cloned and the proteins expressed from a vector derived from Potato virus X (PVX). The expressed HC contained six N-terminal histidine residues to facilitate purification by metal affinity chromatography. Approximately 2–4 µg/g of purified HC was obtained from leaves of Nicotiana benthamiana plants systemically infected by recombinant PVX. Preparations of the HC protein derived from PVY ordinary strain (PVYo) assisted aphid transmission of purified particles of PVYo and PVY strain C (PVYc; a naturally occurring non-aphid transmissible strain of PVY which contains a defective HC), as well as Potato aucuba mosaic virus. The HC derived from PVYc contained the Glu-Ile-Thr-Cys (EITC) motif, and mutation of Glu (E) to Lys (K) enabled the mutant PVX-expressed preparations to assist virus transmission by aphids. Expression of HC protein from the PVX vector produced biologically active protein. This approach should facilitate further studies to elucidate more precisely the molecular mechanism of virus transmission by aphids.




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