ANNOUNCEMENTS
Okra (Abelmoschus esculentus L.), a vital vegetable crop grown extensively in tropical and subtropical regions, suffers significant yield losses due to viral diseases, particularly Yellow Vein Mosaic Virus (BYVMV) and Okra Enation Leaf Curl Virus (OELCuV). Accurate and early detection of these viruses is crucial for effective disease management and breeding resistant varieties. This study aimed to standardize an efficient genomic DNA extraction protocol suitable for okra leaves and validate specific primers for the molecular diagnosis of BYVMV and OELCuV using PCR-based techniques.
Various DNA extraction methods were evaluated and optimized based on DNA yield, purity, cost-effectiveness, and compatibility with downstream applications. The optimized CTAB-based protocol consistently yielded high-quality DNA suitable for PCR amplification, even from field-collected, virus-infected samples. Following DNA isolation, previously reported virus-specific primers were tested and validated for their sensitivity and specificity using PCR on both healthy and symptomatic plant samples. The amplified products were visualized via agarose gel electrophoresis, confirming the presence of expected amplicons for BYVMV and OELCuV in infected plants, while healthy controls showed no amplification.
This standardized protocol not only ensures reliable DNA extraction from okra but also supports accurate and rapid molecular detection of BYVMV and OELCuV. The validation of primers underlines their diagnostic potential in routine virus screening and epidemiological surveys. This study provides a foundational step towards integrated disease management and supports future efforts in developing virus-resistant okra cultivars through molecular breeding.
Keywords: Okra, BYVMV, OELCuV, genomic DNA extraction, primer validation, virus diagnosis, PCR, CTAB protocol, plant virology, molecular diagnostics.
