Development of a plant rhabdovirus-based versatile vector for gene function studies in leafhoppers and rice

Plant viral vectors are powerful tools for transient gene overexpression and silencing, enabling rapid functional analysis without the need for genetic transformation. Rice stripe mosaic virus (RSMV) is an emerging plant rhabdovirus transmitted propagatively by the leafhopper Recilia dorsalis. Leveraging its cross-kingdom replication ability, here we report the development of RSMV as a versatile vector for regulatable foreign gene expression and virus-induced gene silencing (VIGS) in rice and its insect vector. We first established an efficient reverse genetics system for RSMV using Nicotiana benthamiana as a model host. Recombinant virus particles recovered from N. benthamiana leaves were infectious to R. dorsalis and efficiently transmitted to rice. RSMV-based vectors stably accommodated at least two foreign genes, totaling up to 3.7 kb, and maintained stable expression across multiple passages. As proof-of-concept, the RSMV-VIGS vector achieved >90% knockdown of a target gene in R. dorsalis, producing near-knockout phenotypes that persisted throughout adulthood, and also induced efficient gene silencing in infected rice plants. Our work enables genetic manipulation of RSMV for molecular studies and provides a robust tool for functional genomics in both rice and insect hosts.