همراهی یک مولکول بتاستلایت با بیماری ناشی از ویروس پالامپور پیچیدگی برگ گوجه‌فرنگی در گیاهان کدوی خورشتی

Tomato leaf curl Palampur virus (ToLCPalV) (genus Begomovirus, Geminiviridae) is one of the most destructive viruses infecting tomato and different cucurbits in central and southern Iran. ToLCPMV is a bipartite begomovirus and infects the main cucurbit crops other than watermelon. In recent years, association of Tomato leaf curl betasatellite (ToLCB) with a number of begomovirus infected plants have been reported. In this research, the association of ToLCB with the natural ToLCPMV infected zucchini squash (Cucurbita pepo L.) was studied. Zucchini squash samples showing mosaic, dwarfing and severe leaf curling were collected from Anbar-Abad farms (southern Kerman province) and total DNA were extracted from infected tissues followed by enrichment of circular DNA molecules of samples by rolling circle amplification (RCA) method using TempliPhi kit. RCA product of samples was digested with several restriction enzymes and the resulting restriction pattern were analyzed. Regard to identical restriction pattern of RCA product of all samples, the K8 sample was chosen and DNA-A of genome of the associated begomovirus was cloned and Sanger sequenced. The presence of DNA-B of ToLCPalV genome was confirmed using PCR and specific primer pair. Due to severe symptoms of zucchini squash samples, possible association of betasatellite molecule with the K8 sample was checked using PCR assay and overlapped primer pair BS-F/BS-R. The expected 1.4 kb in size amplicon was cloned and Sanger sequenced. Analysis of 2754 and 1368 bp in size sequences indicated that they belong to DNA-A of ToLCPMV and ToLCB genomes, respectively, and shared maximum nucleotide identity with other Iranian isolates available in Genbank. ToLCPMV and ToLCB have a wide distribution in crop growing farms in Kerman Province and reported from various plants. In this study, severe symptoms of zucchini squash plants could be attributed to the association of ToLCB with ToLCPMV. However, further studies are required using infectious clone of the virus and betasatellite to confirm this association.