Complete Genome Sequencing and Infectious cDNA Clone Construction of Soybean Mosaic Virus Isolated from Shanxi

Abstract

Soybean mosaic virus (SMV) is the predominant viral pathogen that affects the yield and quality of soybean. The natural host range for SMV is very narrow, and generally limited to Leguminosae . However, we found that SMV can naturally infect Pinellia ternata and Atractylodes macrocephala . In order to clarify the molecular mechanisms underlying the cross-family infection of SMV, we used double-stranded RNA extraction, rapid amplification of cDNA ends polymerase chain reaction and Gibson assembly techniques to carry out SMV full-length genome amplification from susceptible soybeans and constructed an infectious cDNA clone for SMV. The genome of the SMV Shanxi isolate (SMV-SX) consists of 9,587 nt and encodes a polyprotein consisting of 3,067 aa. SMV-SX and SMV-XFQ008 had the highest nucleotide and amino acid sequence identities of 97.03% and 98.50%, respectively. A phylogenetic tree indicated that SMV-SX and SMV-XFQ018 were clustered together, sharing the closest relationship. We then constructed a pSMV-SX infectious cDNA clone by Gibson assembly technology and used this clone to inoculate soybean and Ailanthus altissima ; the symptoms of these hosts were similar to those caused by the virus isolated from natural infected plant tissue. This method of construction not only makes up for the time-consuming and laborious defect of traditional methods used to construct infectious cDNA clones, but also avoids the toxicity of the Potyvirus special sequence to Escherichia coli , thus providing a useful cloning strategy for the construction of infectious cDNA clones for other viruses and laying down a foundation for the further investigation of SMV cross-family infection mechanisms.

Construction and characterization of the infectious cDNA clone of the prevalent Chinese soybean mosaic virus strain SC3

“…After establishing the first infectious clone of the brome mosaic virus (Ahlquist et al 1984), various plant viruses could be recovered from DNA clones that produce the same infectious activity as naturally occurring viruses (Wang et al 2021;Bao et al 2020). The infectious clone is critical for analyzing viral infection cycles such as viral replication, movement, symptom development, host range, and virus-host interactions (Tuo et al 2017).…”

Generation of a Triple-Shuttling Vector and the Application in Plant Plus-Strand RNA Virus Infectious cDNA Clone Construction

“…Owing to its high transformation efficiency, simplicity, and capacity for complex DNA assembly, HR in yeast has been used in high-throughput plasmid construction, genome synthesis, and genome engineering [ 4 ]. HR-based cloning in yeast has already been successfully applied to the construction of several plant RNA virus infectious clones for agrobacteria-based infiltration [ 8 , 9 , 10 ].…”