An Improved Virus-Induced Gene Silencing (VIGS) System in Zoysiagrass

Virus-induced gene silencing (VIGS) is a powerful RNA-silencing technique for transient gene manipulation and functional verification. However, the procedure of its application among dicot and monocot species varies based on types of vectors, plant susceptibility, as well as inoculation methods. Here, we described a simple yet efficient tobacco rattle virus (TRV)-based VIGS system for functional analysis of genes in zoysiagrass (Zoysia japonica cv. “Zenith”), an important warm-season turfgrass species, using Nicotiana benthamiana as an intermediate host. VIGS of the PDS reporter gene resulted in a characteristic photo-bleaching phenotype in majority of the inoculated leaves up to 70% reduction of expression of the endogenous phytoene desaturase (PDS) gene in zoysiagrass. Our protocol provides a fast and efficient toolbox for high-throughput functional genomics in zoysiagrass species, which could potentially be applied to other warm-season turfgrass species.