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Detection and characterization of two strains of Grapevine fanleaf nepovirus in Tunisia
Author(s) -
Fattouch S.,
Acheche H.,
M’hirsi S.,
Marrakchi M.,
Marzouki N.
Publication year - 2005
Publication title -
eppo bulletin
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.327
H-Index - 36
eISSN - 1365-2338
pISSN - 0250-8052
DOI - 10.1111/j.1365-2338.2005.00819.x
Subject(s) - nepovirus , biology , virology , amplicon , restriction fragment length polymorphism , plant virus , virus , genetics , polymerase chain reaction , gene
The nematode‐borne Grapevine fanleaf nepovirus (GFLV) causes severe degeneration of grapevines in vineyards worldwide. In a recent survey of the sanitary status of grapevine plants in north Tunisian vineyards, we were interested to study the polymorphism of GFLV. Purified virus, from mechanically inoculated Chenopodium quinoa , was used to produce anti‐GFLV antiserum, which specifically recognized GFLV in different Tunisian grapevine samples using the DAS‐ELISA technique. Positive samples were subjected to oligoprobe‐RT‐PCR to amplify a 606 bp region of the viral coat protein sequence. PCR products used for RFLP analysis after digestion with endonuclease Alu I produced 3 restriction profiles. RFLP data allowed clear distinction of two GFLV strains in Tunisia. The nucleotide sequence of the PCR‐generated amplicons from each strain was determined showing 93.4% identity at the nucleic acid level and 97.5% similarity at the aminoacid sequence level compared to the previously characterized GFLV‐F13 French isolate. This paper is the first report on molecular variability of GFLV in Tunisia.