Tobacco rattle virus-induced ( and () gene silencing in L.

Virus-induced gene silencing (VIGS) is an attractive tool for determining gene function in plants. The present study constitutes the first application of VIGS in , which has great ornamental and pharmaceutical value, using (TRV) vectors. Two marker genes, () and (), were used to test the VIGS system in . The photobleaching and yellow-leaf phenotypes of the silenced plants were shown to significantly correlate with the down-regulation of endogenous and , respectively ( ≤ 0.05). Moreover, the parameters potentially affecting the efficiency of VIGS in , including the Agrobacterium strain and the inoculation method (leaf syringe-infiltration, sprout vacuum-infiltration and seed vacuum-infiltration), were compared. The optimized VIGS parameters were the leaf syringe-infiltration method, the Agrobacterium strain GV3101 and the growth of agro-inoculated plants at 25°. With these parameters, the silencing efficiency of and could reach approximately 50% in . Additionally, the suitability of various reference genes was screened by RT-qPCR using three candidate genes, and the results demonstrated that glyceraldehyde 3-phosphate dehydrogenase () can serve as a suitable reference for assessing the gene expression levels of VIGS systems in . The proven application of VIGS in and the characterization of a suitable reference gene in the present work will expedite the functional characterization of novel genes in