Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN)

Abstract

Background Virus-induced gene silencing (VIGS) is a reverse genetics technology that can efficiently and rapidly identify plant gene functions. Although a variety of VIGS vectors have been successfully used in plants, only a few reports on VIGS technology in Luffa exist. Results In the present study, a new cucumber green mottle mosaic virus (CGMMV)-based VIGS vector, pV190, was applied to establish the CGMMV-VIGS to investigate the feasibility of the silencing system for Luffa. Phytoene desaturase (PDS) gene was initially selected as a VIGS marker gene to construct a recombinant vector. Plants infected with Agrobacterium harboring pV190-PDS successfully induced effective silencing in Luffa, and an effective gene silencing phenotype with obvious photobleaching was observed. To further validate the efficiency, we selected TEN for gene-silencing, which encodes a CYC/TB1-like transcription factor and is involved in tendril development. Luffa plants inoculated with the pV190-TEN exhibited shorter tendril length and nodal positions where tendrils appear are higher compared to those of non-inoculated plants. RT-qPCR showed that the expression levels of PDS and TEN were significantly reduced in the CGMMV-VIGS plants. Moreover, we evaluated the CGMMV-VIGS efficiency in three cucurbits, including cucumber, ridge gourd, and bottle gourd. Conclusion We successfully established a CGMMV-based VIGS system on ridge gourd and used marker genes to identify the feasibility of the silencing system in Luffa leaves and stems.

Genome-Wide Analysis of C-Repeat Binding Factor Gene Family in Capsicum baccatum and Functional Exploration in Low-Temperature Response

“…VIGS serves as an approach to rapidly validate the gene function [ 69 ] and has been extensively applied in diverse plants, like tomato, tobacco, Arabidopsis , maize, loofah, cotton, and more [ 70 , 71 , 72 , 73 ]. The effectiveness of the VIGS system varies among plants due to different factors, such as virus–host interaction, plant growth status, and virus invasion [ 74 ].…”

Applications of Virus-Induced Gene Silencing in Cotton

“…The PDS gene is involved in the biosynthesis of carotenoids, and it has been reported in many plant species. It can serve as a visible marker to evaluate the silencing efficiency, as its knockdown in plants can lead to an albino phenotype, due to the lack of chlorophyll [ 34 ]. CLA1 encodes the 1-deoxy-Dxylulose 5-phosphate synthase related to chloroplast development, and it is highly conserved in various plants [ 35 ].…”