Molecular and biological characterization of Chilli leaf curl virus and associated betasatellite infecting Cucurbita max
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Molecular and biological characterization of Chilli leaf curl virus and associated betasatellite infecting Cucurbita maxima in Oman M. S. Shahid1
Received: 23 October 2019 / Accepted: 20 May 2020 Indian Virological Society 2020
Abstract During a survey in February 2016, leaf curl disease symptoms were witnessed in Cucurbita maxima plants in Al-Batina commercial farm in Oman. Symptoms exhibited were characteristic of begomovirus infection as leaf curling, yellowing, and color breaking followed by mosaic pattern. The transmission electron microscopy confirmed the presence of typical twinned geminate typical of Geminate virus particles. Rolling circle amplification (RCA) was employed to characterize the unknown causal agent of C. maxima disease. In molecular identification RCA produced nearly 2.8 and 1.4 kb DNA molecules corresponding to begomovirus and satellite molecules, cloned and sequenced them. In Blast, species demarcation tool and phylogenetic analysis revealed the begomovirus and satellite isolates were determined as Chilli leaf curl virus (ChLCV) and tomato leaf curl betasatellite (ToLCB). In biological analysis by agrobacterium mediated inoculation, ChLCV displayed upward leaf curling and vein swelling symptoms in Nicotiana benthamiana plants; however, in presence of ToLCB enhanced downward leaf curling and crumpling symptoms were revealed. This study provides the first evidence that ChLCV and ToLCB caused leaf curl disease of C. maxima in Oman. Keywords Geminiviridae Begomovirus Chilli leaf curl virus Tomato leaf curl betasatellite Cucurbita maxima
& M. S. Shahid [email protected] 1
Department of Plant Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, 123 Al-Khod, Oman
Plant-infecting single stranded DNA viruses (ssDNA) belongs to genus begomoviruses (family: Geminiviridae) are important pathogens infecting mostly dicotyledonous plants. They are transmitted by whiteflies (Bemisia tabaci) vector and have either a DNA-A or DNA-A and DNA-B genomic components. The DNA-A contain six open reading frames (ORFs), which encode protein for replication, encapsidation and movement. Four (AC1–AC4) ORFs are located in the complementary sense. The AC1 encodes for a Rep (replication associated protein) and AC2 for a TrAP (transcriptional activator protein) while the protein encoded by AC3 is the Ren (replication enhancer protein) whereas the protein encoded by AC4 functions as a suppressor of RNA silencing. The other two ORFs (AV1 and AV2) are located in the virion sense, where AV1 codes for coat protein and the AV2 for a protein of unclear function. Similarly, DNA-B component also contains two ORFs (BC1 in complimentary and BV1 in virion sense). BC1 and BV1 detached by an intergenic region (IR) approximately 300 nucleotides (nt) that contains fundamental elements required for the replication and transcription of the viral genome [2]. Mostly monopartite single stranded DNA (ssDNA) viruses reported from Asia, Europe, Middle East and Australia have been sho
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