2024, Vol. 4, Issue 2, Part A
Development of a PCR-based method for the detection and identification of tobacco mosaic virus and potato virus y in tobacco
Author(s): Reward Muzerengwa, Norman Muzhinji, Dahlia Garwe, Tawanda Jonathan Chisango, Makomborero Nyoni and Frank Magama
Abstract: Morphological techniques have been traditionally relied upon for the detection of Potato Virus Y (PVY) and Tobacco Mosaic Virus (TMV), the two major economic significant viruses infecting tobacco in Zimbabwe and globally. However, morphological methods are subjective and unreliable as they are influenced by abiotic factors and phytotoxicity, leading to misdiagnosis. Accurate, sensitive, and timely diagnosis is crucial for effective management of these viruses to reduce yield losses. The advent of PCR-based methods has revolutionised pathogen diagnostics by providing robust diagnostic methods to support disease management strategies that can prevent yield losses. This study describes a reverse transcriptase- Polymerase Chain Reaction (RT-PCR) protocol developed for the identification of PVY and TMV in tobacco in Zimbabwe. Internal specific primer pairs amplified 480 bp of the coat protein for PVY, 420 bp for the protein movement gene, and 496 bp for the virus genome of TMV. The effectiveness and reliability of the assays were analysed by sensitivity comparisons of double-stranded RNA extraction method (dsRNA), Double Antibody Sandwich-Enzyme Linked Immunosorbent Assay (DAS-ELISA) and RT-PCR conducted at weekly intervals after viral infection of tobacco plants. DsRNA was the least effective detecting PVY after three weeks of infection. DAS-ELISA exhibited more sensitivity than dsRNA, detecting viruses after one week of infection up to six weeks for PVY and three weeks for TMV. However, RT-PCR consistently detected viral infection throughout the duration of the experiment. A cost-benefit analysis of dsRNA, ELISA and RT-PCR was performed, and RT-PCR was found to be slightly more expensive than ELISA and significantly more expensive than dsRNA. Despite the cost differential, the superior sensitivity and reliability of RT-PCR make it the recommended method of detection for improved management of viral diseases in tobacco.
DOI: 10.22271/27893065.2024.v4.i2a.85Pages: 15-21 | Views: 873 | Downloads: 301Download Full Article: Click Here
How to cite this article:
Reward Muzerengwa, Norman Muzhinji, Dahlia Garwe, Tawanda Jonathan Chisango, Makomborero Nyoni, Frank Magama.
Development of a PCR-based method for the detection and identification of tobacco mosaic virus and potato virus y in tobacco. Int J Plant Pathol Microbiol 2024;4(2):15-21. DOI:
10.22271/27893065.2024.v4.i2a.85