RT-PCR-RFLP for genetic diversity analysis of Tunisian Grapevine fanleaf virus isolates in their natural host plants.
Genetic diversity was characterized in 20 isolates of Grapevine fanleaf virus (GFLV) recovered from naturally infected grapevine plants (Vitis vinifera) in the North of Tunisia. Viral RNAs were isolated by oligoprobe capture, and a 605 bp fragment containing a part of the viral coat protein gene was amplified by RT-PCR. Sequence variation among isolates was characterized by restriction fragment length polymorphism (RFLP) analysis and confirmed by sequencing. The GFLV infections are found as a complex mixture of closely related genomes. In further studies, RFLP analyses of virus isolates using AluI showed that GFLV populations in Tunisian vineyards consist of two restrictotypes corresponding to distinct sub-populations Sp1 and Sp2. The relative field distribution of these sub-populations showed that Sp2 was more abundant. Individual genomes were recovered by cloning the RT-PCR products. The sequences were found to vary from each other by as much as 11%. Cloning from mixed infections showed that Sp2 are also predominant.
Fattouch
Sami
S
Laboratory of Biological Engineering, Institut National des Sciences Appliquées et de Technologie, Centre Urbain Nord, BP No. 676, Tunis 1080, Tunisia. Sami.Fattouch@insat.rnu.tn
Acheche
Hajer
H
M'hirsi
Sonia
S
Mellouli
Lotfi
L
Bejar
Samir
S
Marrakchi
Mohamed
M
Marzouki
Najib
N
D014644:Genetic Variation; D016679:Genome, Viral; D017860:Nepovirus / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019685:Rosales / Q000821:virology*