SEROLOGICAL AND MOLECULAR DETECTION OF COWPAE MOSAIC VIRUS INFECTING COWPAE IN IRAQ

Abstract

This study was conducted to detect CPMV in infecting cowpea plantes at Plant Protection Department/College of Agriculture, University of Baghdad. Symptomatic cowpea plants collected from fields in Baghdad and Diyala Provinces were tested by Emzyme-linked immunosorbent assay (ELISA), reverse transcription- polymerase chain reaction RT-PCR and Immuno-capture reverse transcription-polymerase chain reaction (IC-RT-PCR) using commercial kits. ELISA approach could detect Cowpea mosaic virus CpMV in tested samples but with virus concentration lower than the positive control supplied with the kit, indicating a unique virus strain. RNA from samples showed the highest CpMV concentration were extracted by direct release and used in ICPT-PCR using como2 a comovirus specific primer set. cDNA was synthetized using como2 revers primer. RT-PCR confirmed the detection of comoviruses when amplified 200 bp DNA fragments from CpMV infected samples. Sequence analysis confirmed CpMV detection when compared with equivalent GenBank sequences. The sequence obtained shared 94.1% maximum nucleotide identity with RNA dependent RNA polymerase (RdRp) gene of CpMV from Egypt (Acc. No. KT438619 and X00206). Both sequence analysis and IC-RTPCR indicated that virus isolate detected could be an Iraq divergent strain of CpMV.