Construction of an Agroinfectious Clone of a Korean Isolate of Sweet Potato Symptomless Virus 1 and Comparison of Its Infectivity According to Agrobacterium tumefaciens Strains in Nicotiana benthamiana

Abstract

Sweet potato symptomless virus 1 (SPSMV-1) is a single-stranded circular DNA virus, belonging to the genus <i>Mastrevirus</i> (family <i>Geminiviridae</i>) that was first identified on sweet potato plants in South Korea in 2012. Although SPSMV-1 does not induce distinct symptoms in sweet potato plants, its co-infection with different sweet potato viruses is highly prevalent, and thus threatens sweet potato production in South Korea. In this study, the complete genome sequence of a Korean isolate of SPSMV-1 was obtained by Sanger sequencing of polymerase chain reaction (PCR) amplicons from sweet potato plants collected in the field (Suwon). An infectious clone of SPSMV-1 (1.1-mer) was constructed, cloned into the plant expression vector pCAMBIA1303, and agro-inoculated into <i>Nicotiana benthamiana</i> using three <i>Agrobacterium tumefaciens</i> strains (GV3101, LBA4404, and EHA105). Although no visual differences were observed between the mock and infected groups, SPSMV-1 accumulation was detected in the roots, stems, and newly produced leaves through PCR. The <i>A. tumefaciens</i> strain LBA4404 was the most effective at transferring the SPSMV-1 genome to <i>N. benthamiana</i>. We confirmed the viral replication in <i>N. benthamiana</i> samples through strand-specific amplification using virion-sense- and complementary-sense-specific primer sets.

A simple and efficient Agrobacterium tumefaciens mediated transgenic system for tetraploid potato cultivar Desiree

“…According to Vinterhalter et al (2008), LBA4404 is the most frequent, which has been used for the introduction of binary vectors 13 . In the last published protocol, EHA105 was applied more frequently [14][15][16] .…”