Molecular Confirmation of Plant Viruses Detected by High-Throughput Sequencing (HTS) in Fruit Trees Using RT-PCR

This study aimed to confirm the presence of Citrus Exocortis Viroid (CEVd) in Ficus carica (fig) trees in the Al-Kufa and Al-Hilla regions of Iraq using a molecular diagnostic approach. Initially, high-throughput sequencing (HTS) data suggested the occurrence of CEVd in symptomatic fig leaf samples. To validate these findings, total RNA was extracted from field-collected samples and subjected to reverse transcription followed by polymerase chain reaction (RT-PCR) using CEVd-specific primers. The RT-PCR results confirmed the viroid’s presence in 30% of the tested samples from Al-Kufa and 28% from Al-Hilla. This molecular confirmation not only supports the HTS data but also marks the first RT-PCR-based detection of CEVd in fig trees in Iraq. The results highlight the need for routine molecular surveillance and suggest a potential involvement of CEVd in fig mosaic disease development in the studied regions.