A highly efficient Agrobacterium-mediated infectious system for Sweet potato leaf curl virus and a deltasatellite-based VIGS vector

Abstract

Plant virus disease is one of the major threats to the yield and quality of sweetpotato (Ipomoea batatas [L.] Lam.), the seventh most important food crop in the world in terms of production. Sweepoviruses are a group of monopartite geminiviruses (genus Begomovirus, family Geminiviridae) that infect Ipomoea plants and cause significant damage. However, investigations on the mechanisms of sweepovirus infection are scarce due to the low efficiency of the current infectious system. In this work, we used the genomic sequence of Sweet potato leaf curl virus (SPLCV) to construct an effective infectious clone, SPLCV-1.01. We optimized the inoculation procedure in Nicotiana benthamiana by improving the Agrobacterium virulence using an induction medium. Finally, we established a highly efficient agroinoculation protocol for SPLCV on sweetpotato. Furthermore, a simple and reliable “Agro-soaking” method was developed for SPLCV on sweetpotato and Ipomoea aquatica. The “Agro-soaking” method also worked well with Tomato yellow leaf curl China virus (TYLCCNV) and Beet severe curly top virus (BSCTV) on tomato. Based on the “Agro-soaking” method and a deltasatellite, the SBG51 VIGS vector worked with the SPLCV-1.01 infectious clone and silenced two endogenous genes IbCHL (magnesium-chelatase subunit) and IbPDS (phytoene desaturase) in two weeks. Moreover, the VIGS system identified IbADK1 (adenosine kinase 1) as a positive regulator to dampen the SPLCV accumulation in sweetpotato. Our work provides a practical method for studying the interaction between sweetpotato and SPLCV and the functional studies of genes in sweetpotato.