(GFLV) is one of the most severe virus diseases of grapevines, causing fanleaf degeneration that is transmitted by This paper aims to isolate species from Tunisian vineyard soil samples and assess their ability to acquire and transmit GFLV under natural and controlled conditions. Based on morphological and morphometric analyses, Tunisian dagger nematodes were identified as and These results were confirmed with molecular identification tools using species-specific polymerase chain reaction primers. The total RNA of GFLV was extracted from specimens of and amplified based on real-time polymerase chain reaction using virus-specific primers. Our results showed that could acquire and transmit the viral particles of GFLV. This nepovirus was not detected in , under natural conditions; however, under controlled conditions, this nematode was able to successfully acquire and transmit the viral particles of GFLV.