Plant leaves used in this study were collected from Nicotiana benthamiana plants, inoculated with ToLCNDV, growing under glass house conditions at National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan.
benthamiana plants infected with ToLCNDV were subjected for the IC-PCR detection of ToLCNDV.
Stephen Winter) were used for immunocapture of ToLCNDV.
The degenerate primers CLCV1/CLCV2 and specific primers (PadCPPVXF/PadCPPVXR) were used for detection of ToLCNDV (Table I).
The amino acid sequences of CPs of ToLCNDV, ACMV and TYLCV were retrieved from the database with accession numbers AAA92807, CCH63378 and CAL64776, respectively, and used to unravel the structural aspects of these proteins.
Detection of ToLCNDV by using TYLCV- and ACMV-CP antisera
The possibility of detection of ToLCNDV DNA-A from plant extract was examined by immunocapture PCR.
Such interactions are also suggestive of sharing antigenic relationship between particle protein of ToLCNDV and TYLCV-CP.
Comparison of conventional PCR with IC-PCR for ToLCNDV detection
A serological method for ToLCNDV detection based on immunocapturing with TYLCV-CP antiserum followed by PCR amplification was established.
Amino acid pairwise alignment of CP of ToLCNDV, ACMV and TYLCV showed that ToLCNDV shared more sequence similarity with ACMV than TYLCV.