DEVELOPMENT OF A MOLECULAR ASSAY FOR THE DETECTION OF TOMATO MOSAIC VIRUS IN SRI LANKA
B. M. V. S. BASNAYAKE *
Plant Virus Indexing Center, Gabadawaththa, Homagama, Sri Lanka.
A. D. R. P. ALUTHGE
Plant Virus Indexing Center, Gabadawaththa, Homagama, Sri Lanka.
A. W. P. AMARAJEEWA
University of Sri Jayawardenapura, Gangodawila, Nugegoda, Sri Lanka.
*Author to whom correspondence should be addressed.
Abstract
Virus diseases in tomato cultivations are one of the limiting factors for large-scale production in Sri Lanka. Tomato mosaic virus(ToMV) is an economically important virus disease as tomato is highly susceptible to the disease. Similarity of symptoms has made identification difficult. Developing a reliable indexing method for the identification of ToMV is aimed in this study. Saps extracted from infected leaves were subjected to Enzyme Linked Immunosorbant Assey (ELISA) and positive samples were used in the mechanical inoculation. After symptom expression the samples were subjected to ELISA and the disease was confirmed. Viral RNA was extracted from confirmed disease samples and used in Reverse Transcription Polymerase Chain Reaction (RT-PCR) with specific primers ToMV 1F/ 1R. A gradient PCR detected the annealing temperature as 53°C. The protocol was optimized and the optimized protocol was used to detect the disease in samples collected from several districts in Sri Lanka. The study revealed the presence of ToMV in Sri Lanka and a molecular based detection technology wasdeveloped. This is the first record of a molecular based detection of ToMV in Sri Lanka.
Keywords: Tomato mosaic virus (ToMV), Reverse Transcription Polymerase chain reaction (RT-PCR), Enzyme Linked Immunosorbant Assey (ELISA)