Identification of protein interactions of grapevine fanleaf virus RNA-dependent RNA polymerase during infection of Nicotiana benthamiana by affinity purification and tandem mass spectrometry

The RNA-dependent RNA polymerase (1E Pol ) is involved in replication of grapevine fanleaf virus (GFLV, Nepovirus , Secoviridae and causes vein clearing symptoms Nicotiana benthamiana . Information on protein 1E interaction with other viral host proteins scarce. To study biology, three GFLV infectious clones, i.e. GHu (a symptomatic wild-type strain), GHu-1E K802G (an asymptomatic mutant) F13 were engineered fused to a V5 epitope tag at the C-terminus. Following Agrobacterium tumefaciens -mediated delivery clones N. extraction seven dpi, when optimal :V5 accumulation was detected, two six plant putative partners V5-tagged identified for by affinity purification tandem mass spectrometry. This provides insights into interactome during systemic infection lays foundation validation work.