FIRST REPORT OF CHILLI VEINAL MOTTLE VIRUS IN TOMATO IN PAKISTAN

Abstract

Chilli veinal mottle virus (ChiVMV, genus Potyvirus ) infects several solanaceous species worldwide (Zhao et al ., 2014). In April 2014, a total of 20 tomato leaf samples with symptoms of mosaic, necrosis and mottling were collected from the research farm of PMAS-Arid Agriculture University Rawalpindi. All disease samples were screened for the presence of potyvirus infection by indirect plate-trapped antigen (PTA)-ELISA using “Poty group test” kit (Bioreba AG, Switzerland). Only 12 samples were positive for potyvirus infection. These samples were further tested for Potato virus Y (PVY), Pepper veinal mottle virus (PVMV) (Bioreba AG) and ChiVMV (Loewe Biochemicals, Germany) using virus specific DAS- ELISA. Of 12 potyvirus confirmed samples, two were positive for ChiVMV and remaining ten were positive for PVY. No reaction was observed with antibodies against PVMV. The ChiVMV ELISA-positive samples were further confirmed by RT-PCR using ChiVMV specific primers Poty3 and Oligo(dT) (Tsai et al ., 2008). Both PCR amplicons were purified using QIAquick® PCR purification kit (Qiagen) and subsequently sequenced in both orientations . A product 759 nucleotides in size (partial CP gene including 3’UTR) was obtained (both clones were 100% identical) and the sequence of ChiVMV isolate tomato (AARTPK) was deposited in GenBank (Accession No. KT876048). BLASTn revealed >90% sequence identity with ChiVMV isolates from India (JN624776, JN692501), China (HQ218936, KF738253, KC711055, JX088636) and Pakistan (KJ472764). This destructive virus has been previously reported from chilli pepper in Pakistan (Shah et al ., 2009). To the best of our knowledge, this is the first confirmed report of ChiVMV from tomato in Pakistan.