A temperature-controlled amplicon system derived from Plum pox potyvirus

Authors: Dujovny, Gabriela; Valli, Adrián; Calvo, María; García, Juan Antonio

Source: Plant Biotechnology Journal, Volume 7, Number 1, January 2009 , pp. 49-58(10)

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Abstract:

Summary

The control of replication can facilitate a viral amplicon to reach high expression levels by enabling the virus to escape host defence mechanisms and reducing the deleterious effects of viral infection. We have developed a novel system to regulate amplicon expression by controlling the temperature of plant growth. Nicotiana benthamiana plants were transformed at two different temperatures with a cDNA copy of the Plum pox potyvirus genome harbouring the open reading frame 2 of Porcine circovirus 2 between the nuclear inclusion protein b and coat protein coding sequences. Although transformation at 27 °C mainly yielded nonexpressing amplicons, lines with a tight control of amplicon expression were obtained. Viral replication was not detected in these plants when germinated at 28 °C, but was observed when the plants were shifted to 20 °C. In lines transformed at 24 °C, although the amplicon was expressed at 28 °C, viral accumulation was low and caused only minor growing defects. Viral replication was enhanced in these plants by shifting the temperature to 20 °C; under such conditions, the amplicon reached higher and more persistent expression levels than in plants transformed at 27 °C. These results demonstrate the utility of temperature regulation to control viral amplicon expression.

Keywords: amplicon; controlled expression; expression vector; Plum pox virus; potyvirus; virus vector

Document Type: Research article

DOI: http://dx.doi.org/10.1111/j.1467-7652.2008.00373.x

Publication date: 2009-01-01