Production of monoclonal antibodies against synthetic peptides of the N‐terminal region of Potato virus Y coat protein and their use in PVY strain differentiation

Abstract

Antibodies were prepared against two synthetic peptides, P19 and P11, derived from the coat protein N-terminal region of two pepper isolates of Potato virus Y from Tunisia (PVY-P21 and PVY-P2, respectively). The peptides were selected by comparing the predicted amino acid sequences of three pepper and four potato PVY isolates on the basis of their polymorphism and hydrophilicity. Sera with high titres were only obtained against P19. Three MAbs, raised in response to P19, reacted with the homologous virus (PVY-P21) in TAS-ELISA. When tested against a broad range of PVY isolates and related viruses, MAb 3C5 proved to be PVY species specific, whereas MAbs 8A4 and 1D6 reacted specifically with standard isolates of PVY O , PVY C and PVY N -W strains, but not with other PVY isolates. Consequently, epitope(s) recognized by 8A4 and 1D6 MAbs may be specific to a PVY group comprising all serologically PVY non-N isolates. Surprisingly, and unlike isolate PVY-P21, many Tunisian field pepper isolates did not carry this epitope(s), thus revealing serological heterogeneity within the PVY pepper group. As PVY is one of the most economically important plant pathogens in a range of crops, including pepper, these MAbs will provide a useful tool for practical diagnosis and strain identification of PVY.

Mycoviruses of filamentous fungi and their relevance to plant pathology

“…One obvious outcome of the ability to detect, clone and sequence mycovirus genomes has been the discovery of a much greater diversity of viruses in fungi, including those with ssRNA genomes. In some instances molecular data will provide an avenue to revisit and more effectively use older technologies such as the production of virus antibodies to synthetic peptides based on genome sequence of virus proteins, as has been achieved for the plant virus Potato virus Y (Ounouna et al ., 2002). Although dsRNA profiles have been used for several large‐scale surveys of mycoviruses (Table 3), the specific identity of the viruses detected is often unknown or uncertain.…”

Molecular and Biological Characterization of Potato virus Y Isolates from Vietnam

“…There are several different MAb's with recognition motifs different from those of Bioreba's N‐MAb that can be used to differentiate between NTN and Wilga isolates (Ounouna et al. ; Chikh Ali et al. ; Ranki et al.…”

“…There are several different MAb's with recognition motifs different from those of Bioreba's N-MAb that can be used to differentiate between NTN and Wilga isolates (Ounouna et al 2002;Chikh Ali et al 2007;Ranki et al 2008;Nikolaeva et al 2012). This might cause confusion as a single point mutation in the rec-ognition motifs in the CP gene would change the classification of the strains but leave the biological features of the isolates unchanged.…”

The acquisition of molecular determinants involved in potato virus Y necrosis capacity leads to fitness reduction in tobacco plants

“…Xanthi leaves. Since the description of these two main PVY groups, numerous biological (Jones, 1990;Valkonen, 1997), serological (Rose & Hubbard, 1986;Cerovska, 1998;Ounouna et al, 2002) and molecular (Weidemann & Maiss, 1996;Glais et al, 2005;Kogovsek et al, 2008;Rolland et al, 2008) diagnostic tools have been developed for the detection of PVY isolates and the description of their diversity. According to their characteristics (specificity, sensitivity and accuracy), these tools have been used to describe PVY and/or to estimate the proportion of necrotic isolates in natural populations (Crosslin et al, 2006).…”