Coat Protein Gene: A key tool for Yam Mild Mosaic Virus diagnosis in greater yam

Abstract

Destruction of infected plants and use of virus-free planting materials are the common control measuresused for preventing viral infections. These practices are inadequate to combat the virus attack in manyincidents. Early detection of infection is an effective way to manage the systemic primary spread ofviruses. Infection caused by Yam mild mosaic virus (YMMV), one of the major viruses in yams (Dioscoreaspp) is difficult to detect in the early stages. Symptoms include mild mosaic, leaf mottling, and chlorosisand leaf distortion. In the present study, PCR was employed for the amplification of YMMV coat protein(CP) gene for diagnosing the virus from greater yam (Dioscorea alata L.) leaves. The specific primerpair was designed and validated for CP gene amplification, which yielded an amplicon of 810 bp inYMMV positive samples. Subsequent cloning in pUC18 vector and sequencing confirmed the presenceof the full coat protein (CP). In addition to PCR-based diagnostic method, the accomplished isolationand characterization process opens avenues for generating virus-specific polyclonal antibodies throughthe utilization of the expressed coat protein. These antibodies can be further employed in serologicaltechniques.