2004 Jul 8;4:26. doi: 10.1186/1471-2180-4-26. Privacy 2019). One year later, the recombinant infectious clones of vesicular stomatitis virus (VSV) were constructed (Pattnaik et al. 2012; Jarugula et al. Major breakthroughs in reverse genetics systems of plant non-segmented and segmented NSR viruses were just achieved in recent 5 years through various strategies, such as agroinfiltration, minireplicon systems, insect transmission and airbrush inoculation assays. The BYSMV genome encodes ten proteins including five structural proteins and five accessory proteins in the order of 3′–N–P–P3–P4/P5–P6–M–G–P9–L–5′ (Yan et al. 2018; Jiang et al. 2019).  |  All these constructs can support recombinant RRV infections in A. thaliana and N. benthamiana plants by agroinfiltration and in roses by the airbrush inoculation approach (Pang et al. Tomato spotted wilt virus is a segmented NSR virus and belongs to the genus Orthotospovirus in the family Tospoviridae (Kormelink et al. 2013). A human papillomavirus 8 E7 protein produced in plants is able to trigger the mouse immune system and delay the development of skin lesions. -, Proc Natl Acad Sci U S A. This site needs JavaScript to work properly. HHS  |  2019; Feng et al. Infectious DNA clone technology and inoculation strategy for Rose Rosette Virus that includes all seven segments of the negative-strand RNA genome. J Biotechnol. In addition, the BYSMV vector could express a RFP reporter protein to tract the BYSMV infections and VIGE sites (Gao et al. Therefore, rescue of infectious plant NSR viruses from cDNA clones strictly requires the core replication proteins together with their genome RNAs which can assemble into nucleocapsid (NC) complexes as minimal infectious units. developed an effective mechanical method by using an airbrush to deliver agrobacterium cultures (Pang et al. Our studies clearly provide new protocols for developing reverse genetics systems of plant NSR viruses infecting monocot plants and arthropod vectors. 2020;6:773–9. 2020). In addition, BYSMV-mediated overexpression of B2, a suppressor of RNA silencing in insect, could improve the BYSMV infections in SBPHs, which indicates that BYSMV expression vectors can be applied in planthoppers (Gao et al. Annu Rev Phytopathol. J Exp Bot. 2019). have successfully established infectious TSWV entirely from cDNA clones in plants. 2019; Gao et al. Please enable it to take advantage of the complete set of features! f Inoculation by an airbrush was effective in delivering agrobacterium cultures containing RRV vectors to A. thaliana, N. benthamiana and roses. For instance, BSMV, wheat streak mosaic virus (WSMV), soilborne wheat mosaic virus (SBWMV), and foxtail mosaic virus (FoMV) have been engineered as VOX vectors to express foreign proteins in wheat, barley, oat, and maize plants, respectively (Choi et al. Open Access . In our recent studies, BYSMV, a plant cytorhabdovirus, was developed as VOX vectors that can simultaneously express three foreign proteins (GFP, RFP, and CFP) in both monocot plants and insect vectors (Gao et al. Nevertheless, the reported success of genome editing is based on Cas9 transgenic plants, because positive strand RNA viruses-based vectors are incapable of carrying the large Cas9 genes. 2018;177:1352–67. Plant Biotechnol J. In addition, green fluorescent protein (GFP), red fluorescent protein (RFP), and/or chloramphenicol acetyltransferase (CAT) reporter genes replaced the N and P genes to visualize MR transcription and replication in the cells (Fig. b The minireplicon (MR) containing a double cauliflower mosaic virus 35S (2 × 35S) promoter, exact termini of the virus, gene junctions, and reporter genes could replicate in conjunction with the ectopic expression of viral core proteins and VSRs, which could validate whether the core replicate proteins are functional in viral transcription and replication. As described above, plant rhabdoviruses could stably express multiple and large foreign proteins, which expends the research into expression of both the sgRNA and the Cas9 nuclease in one virus vector. Top 10 plant viruses in molecular plant pathology. 2019). 2011;12:938–54. Part of Development of Beet necrotic yellow vein virus-based vectors for multiple-gene expression and guide RNA delivery in plant genome editing. Liu Z, Kearney CM. Drug Discov Today. Currently, Cas9 nucleases and guide RNAs are mainly delivered by transformation to obtain stable transgenic plants or transiently delivery into protoplasts (Cong et al.