title: Development of a new vector using Soybean yellow common mosaic virus for gene function study or heterologous protein expression in soybeans.
abstract: A new vector using Soybean yellow common mosaic virus (SYCMV) was constructed for gene function study or heterologous protein expression in soybeans. The in vitro transcript with a 5' cap analog m7GpppG from an SYCMV full-length infectious vector driven by a T7 promoter infected soybeans (pSYCMVT7-full). The symptoms observed in the soybeans infected with either the sap from SYCMV-infected leaves or pSYCMVT7-full were indistinguishable, suggesting that the vector exhibits equivalent biological activity as the virus itself. To utilize the vector further, a DNA-based vector driven by the Cauliflower mosaic virus (CaMV) 35S promoter was constructed. The complete sequence of the SYCMV genome was inserted into a binary vector flanked by a CaMV 35S promoter at the 5' terminus of the SYCMV genome and a cis-cleaving ribozyme sequence followed by a nopaline synthase terminator at the 3' terminus of the SYCMV genome (pSYCMV-full). The SYCMV-derived vector was tested for use as a virus-induced gene silencing (VIGS) vector for the functional analysis of soybean genes. VIGS constructs containing either a fragment of the Phytoene desaturase (PDS) gene (pSYCMV-PDS1) or a fragment of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (RbcS) gene (pSYCMV-RbcS2) were constructed. Plants infiltrated with each vector using the Agrobacterium-mediated inoculation method exhibited distinct symptoms, such as photo-bleaching in plants infiltrated with pSYCMV-PDS1 and yellow or pale green coloring in plants infiltrated with pSYCMV-RbcS2. In addition, down-regulation of the transcripts of the two target genes was confirmed via northern blot analysis. Particle bombardment and direct plasmid DNA rubbing were also confirmed as alternative inoculation methods. To determine if the SYCMV vector can be used for the expression of heterologous proteins in soybean plants, the vector encoding amino acids 135-160 of VP1 of Foot-and-mouth disease virus (FMDV) serotype O1 Campos (O1C) was constructed (pSYCMV-FMDV). Plants infiltrated with pSYCMV-FMDV were only detected via western blotting using the O1C antibody. Based on these results, we propose that the SYCMV-derived vector can be used for gene function study or expression of useful heterologous proteins in soybeans.
authors.affiliation: Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea; Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea.
authors.forename: Seungmo
authors.initials: S
authors.lastname: Lim
authors.affiliation: School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Moon
authors.initials: M
authors.lastname: Nam
authors.affiliation: National Institute of Crop Science, Rural Development Administration, Suwon 441-857, Republic of Korea.
authors.forename: Kil Hyun
authors.initials: KH
authors.lastname: Kim
authors.affiliation: School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Su-Heon
authors.initials: SH
authors.lastname: Lee
authors.affiliation: National Institute of Crop Science, Rural Development Administration, Suwon 441-857, Republic of Korea.
authors.forename: Jung-Kyung
authors.initials: JK
authors.lastname: Moon
authors.affiliation: Department of Applied Biology, Chungnam National University, Daejeon 305-764, Republic of Korea.
authors.forename: Hyoun-Sub
authors.initials: HS
authors.lastname: Lim
authors.affiliation: Department of Herbal Medicine Resource, Kangwon National University, Samcheok 245-710, Republic of Korea.
authors.forename: Myoung-Gun
authors.initials: MG
authors.lastname: Choung
authors.affiliation: Yeongnam Regional Office, Animal and Plant Quarantine Agency, Busan 600-016, Republic of Korea.
authors.forename: Sang-Mok
authors.initials: SM
authors.lastname: Kim
authors.affiliation: Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea; Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea. Electronic address: jsmoon@kribb.re.kr.
authors.forename: Jae Sun
authors.initials: JS
authors.lastname: Moon
mesh_terms: D036022:Capsid Proteins / Q000235:genetics; D017796:Caulimovirus / Q000235:genetics; D015536:Down-Regulation; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D009029:Mosaic Viruses / Q000235:genetics*; D010641:Phenotype; D018515:Plant Leaves / Q000821:virology; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D012367:RNA, Viral / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013025:Glycine max / Q000235:genetics* / Q000378:metabolism / Q000821:virology
chemical_list: D036022:Capsid Proteins; D012367:RNA, Viral; D011994:Recombinant Proteins; C442501:VP1 protein, Foot-and-mouth disease virus
keywords: Protein expression; SYCMV; Soybean; VIGS