First report of cotton leaf curl Gezira virus incidence and the associated betasetellite in marshmallow, okra and sunflower in Iran

The genus Begomovirus is the largest genus of the family Geminiviridae. In this research, a common marshmallow (Malva sp.) sample with the begomovirus-like symptoms including crinkled and curling leaves was collected in Jiroft. Total DNA was extracted using the CTAB method and subjected to next generation sequencing approach. Sequence analysis of NGS data indicated that the common marshmallow sample is infected with the Cotton leaf curl Gezira virus (CLCuGV) and Cotton leaf curl Gezira betasatellite. To confirm the sequence data, the full-length genome of CLCuGV and the associated betasatellite were amplified using rolling circle amplification method and the RCA kit followed by cloning and Sanger sequencing of ~3kb amplicon. Sequence data indicated that genomes of the Iranian CLCuGV isolate and the associated betasatellite are 2761 and 1355 nts in length. To identify other CLCuGV hosts, a sunflower sample showing severe crinkling and cup-shaped leaves and five okra samples with severe leaf curl symptom were collected from greenhouse and/or open farms in Jiroft and Dezful (Khuzestan province), respectively. DNA of samples was extracted and subjected to the PCR assay using primer pair of CLCuGV-F/CLCuGV-F followed by detection of 730bp in length amplicon. The associated betasatellite with the okra and sunflower samples was detected by RCA and sequencing of the resulted amplicon.