siRNA Silencing of PVX Coat Protein Gene Affects Accumulation of Viral RNA in Potato and Tobacco Plants

Research Article
siRNA Silencing of PVX Coat Protein Gene Affects Accumulation of Viral RNA in Potato and Tobacco Plants

A.M. Soliman, B.N. Barsoum, G.G. Mohamed, A.A. Rezk, A.E. Aboul-Ata and H.M. Mazyad

International Journal of Virology, 2008, 4(1), 14-25.

Abstract

This study, aims at determination of efficiency of micro interfering RNA (miRNA) to develop ability of virus resistance against Egyptian PVX isolate (PVX-Eg2) in both potato (Solanum tuberosum L. cv. Spunta) and tobacco (Nicotiana benthamiana). RNA constructs of Sense (PVX-Eg2cpVs), antisense (PVX-Eg2cpCs) and sense/antisense were designed, cloned and sub- cloned for gene transfection using Agrobacterium inoculation technique. Two to 3 leaf-stage seedlings of potato (Solanum tuberosum L. cv. Spunta) and tobacco (Nicotiana benthamiana) were inoculated with the three previous constructs. The construct-treated plants were mechanically inoculated with the PVX-Eg2 isolate. Bioassay and PCR amplification have been able to evaluate transfected-plant resistance against PVX-Eg2 that is caused by siRNA of PVX-Eg2cp. PCR amplification has been able to detect PVX viral genome in all challenged plants those infiltrated with either pFGC5491 vector without insert, or with sense construct and also with antisense construct. Bioassay has confirmed same previous statement. Nine out of 10 sense/antisense-transfected potato plants were negatively reacted with both bioassay and PCR amplification. Same negative reaction has been viewed using both bioassay and PCR for sense/antisense transfected-tobacco plants. Seven out of 10 proved they are PVX-Eg2 resistant.

ASCI-ID: 45-1

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