Molecular Detection and Antiserum Preparation of Begomoviruses Infecting Zucchini Squash in Iraq
AbstractBegomoviruses infecting zucchini squash were investigated. Leaf samples were collected from zucchini squash growing areas in Baghdad (Jhadryaa and Yusufiyah), Babylon (Jibela and Mahmudiyah) and Diyala (Khan Bani Saad) Provinces. Samples were screened for the presence of begomoviruses using polymerase chain reaction (PCR) and Deng genus specific primers. Sixteen out of 40 samples were begomovirus positive. Sequence analysis confirmed the detection of Tomato leaf curl Palampur virus (TLCPALV) Begomovirus solanumpalampurense, Squash leaf curl virus (SLCuV) B. cucurbitapeponis and Tomato yellow leaf curl virus (TYLCV) B. coheni in zucchini squash and Datura stramonium samples. Co-infections of B. solanumpalampurense, and B. cucurbitapeponis, B. solanumpalampurense and B. coheni were detected in most samples suggesting begomovirus mixed infections are common in the investigated zucchini squash fields. Datura stramonium can be an alternatives host to B. solanumpalampurense and B. coheni. Total protein extracted from begomovirus infected samples were injected into lab rats to prepare begomovirus specific antibodies. The resultant polyclonal antibodies were labeled using horseradish peroxidase and enzyme linked immune assay (ELISA) was performed. Antibodies prepared could detect begomovirus in infected samples scoring 1.247 highest absorbance value at 405 nm compared to 1.198 of slandered TYLCV antigen. Proteins extracted from begomovirus co-infected tissues can be used to immunize rats for the production of beganovirus specific polyclonal antibodies.