Virus-Induced Gene Silencing in Lilies Using Cucumber Mosaic Virus Vectors.

Virus-induced gene silencing (VIGS) systems are effective for rapid analysis of gene functions in plants that require a long period of growth such as Lilium. We successfully developed a VIGS system using the cucumber mosaic virus (HL strain, CMV-HL) vector to induce RNA silencing of the L. leichtlinii phytoene desaturase gene (LlPDS), where at 30 days postinoculation (dpi), photo-bleaching was observed in the upper leaves of L. leichtlinii, and at 57 dpi, white regions appeared on flower tepals that accumulate orange carotenoids. This vector spreads in bulbs, and it could induce silencing on emerged shoots in the following year. The CMV-HL vector can be easily constructed by insertion of a 30-60 nt fragment into the cloning site of the RNA3 genome. In this chapter, we describe how to use the CMV-HL vector system in the context of Lilium plants.