Establishment and optimization of a tobacco rattle virus -based virus-induced gene Silencing in Atriplex canescens.

UNLABELLED

, a halophytic model plant exhibiting remarkable tolerance to abiotic stress, possesses valuable genetic resources for crop improvement. However, the absence of a reliable and stable transformation systems has impeded the functional characterization of candidate genes in this species. In this study, we established an efficient virus-induced gene silencing (VIGS) system in by constructing a tobacco rattle virus (TRV)-based vector targeting the phytoene desaturase gene (), whose silencing induces an easily recognizable photobleaching phenotype. Through comparative analyses of inoculation materials and methods for VIGS implementation in , we found that inoculating germinated seeds with suspension (OD = 0.8) carrying TRV2: vectors through vacuum-assisted agroinfiltration (0.5 kPa, 10 min) further achieving an average silencing efficiency of approximately 16.4%. Systemic photobleaching phenotypes appeared in newly emerged leaves at about 15 days post-inoculation, accompanied by a 40–80% reduction in transcript abundance, as confirmed by qRT-PCR. Furthermore, we successfully validated this VIGS system by silencing two aquaporin genes ( and ), achieving 60.3–69.5% knockdown efficiency, which confirmed the broad applicability of this system in . In summary, our method provides a reliable reverse-genetics platform that facilitates the study of stress-resistance mechanisms in , effectively overcoming previous technical limitations in genetic research on this species.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1186/s13007-025-01427-z.