Liebenberg A., Freeborough M.-J., Visser C.J., Bellstedt D.U., Burger J.T.
Vitis laboratory, Department of Genetics, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa; Department of Biochemistry, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa
Liebenberg, A., Vitis laboratory, Department of Genetics, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa; Freeborough, M.-J., Vitis laboratory, Department of Genetics, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa; Visser, C.J., Department of Biochemistry, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa; Bellstedt, D.U., Department of Biochemistry, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa; Burger, J.T., Vitis laboratory, Department of Genetics, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa
Grapevine fanleaf virus (GFLV) is responsible for severe fanleaf degeneration in grapevines of all major wine producing regions of the world, including South Africa. In order to successfully control the spread of the virus, specific and reliable diagnostic assays are necessary. The genetic variability of 12 GFLV isolates recovered from naturally infected grapevine plants in the Western Cape region of South Africa were characterised. These samples were subjected to RNA extraction, RT-PCR analysis and sequencing of the coat protein gene (2CCP). Sequence identities between different GFLV isolates from South Africa were between 86-99% and 94-99% at the nucleotide and amino acid levels, respectively. Phylogenetic analysis based on the 2CCP gene sequences showed that the South African isolates form two distinct clades or sub-populations. The specificity and sensitivity of three diagnostic techniques (rapid-direct-one-tube-RT-PCR, DAS-ELISA and ImmunoStrips) for the detection of GFLV were analysed to determine the appropriate diagnostic assay for virus infection. Rapid-direct-one-tube-RT-PCR was found to be the most reliable technique for detection. This is the first report on sequence analysis of full-length 2CCP gene cDNA clones of GFLV isolates from South Africa. © 2009 Elsevier B.V. All rights reserved.
amino acid; coat protein; nucleotide; article; cladistics; climbing plant; controlled study; double antibody sandwich enzyme linked immunosorbent assay; gene sequence; genetic variability; grapevine; grapevine fanleaf virus; leaf damage; Nepovirus; nonhuman; nucleotide sequence; phylogeny; priority journal; reverse transcription polymerase chain reaction; RNA extraction; sensitivity and specificity; unindexed sequence; virus diagnosis; Capsid Proteins; DNA, Viral; Enzyme-Linked Immunosorbent Assay; Genetic Variation; Immunoassay; Molecular Sequence Data; Nepovirus; Phylogeny; Plant Diseases; Reverse Transcriptase Polymerase Chain Reaction; South Africa; Vitis; Fioria; Grapevine fanleaf virus; Vitis