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Development of new vector by soybean yellow common mosaic virus for foreign gene expression and virus-induced gene silencing in soybean
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/297911
In this study, we constructed viral vector for soybean by using Soybean yellow common mosaic virus (SYCMV) infecting both Glycine max and Glycine soja. SYCMV-derived viral vector was tested to use as Virus-induced gene silencing (VIGS) vector for functional analysis of soybean genes and as protein expression vector for foreign protein expression. In vitro transcript with 5’ cap analog m7GpppG from a full-length infectious vector of SYCMV driven by T7 promoter was inoculated to soybean to test infectivity of the clone (pSYCMVT7-full). 5’-capped transcript was able to infect soybean plants. The symptoms observed in soybean plants infected by either the vector or the sap from SYCMV-infected leaves were indistinguishable, suggesting that the vector had an equal biological activity shown by virus itself. To further utilize the vector, an additional DNA-based vector was constructed. The full-length cDNA was inserted into a binary vector flanked by CaMV 35S promoter and the nopaline synthase terminator (pSYCMV35S-full). To test if the vector infects soybean and subsequently induces gene silencing, we prepared two constructs containing fragments of Phytoene desaturase (PDS) gene (pSYCMV35S-PDS1) and small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcS) gene (pSYCMV35S-rbcS2) from soybean plant. Plants infiltrated with the constructs through Agrobacterium-mediated method showed distinct symptoms such as photobleaching in plants infiltrated with pSYCMV-PDS1 and pale green or yellowing in plants infiltrated with pSYCMV-rbcS2. In addition, down-regulations of mRNA levels of two genes were confirmed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). To test if the vector can be used for foreign protein expression in soybean plants, we prepared a construct encoding amino acids 135-160 of VP1 FMDV serotype O1 Campos (O1C) (pSYCMV35S-FMDV). Plants infiltrated with the construct through Agrobacterium-mediated method showed that soybean plant infiltrated with pSYCMV35S-FMDV only was detected by Western blotting using O1C antibody. These results support that SYCMV-derived viral vector can be used as VIGS vector or protein expression vector in soybean plants.
