Polyclonal antibodies production against Pepper mild mottle virus affecting pepper plants in Egypt

M. A. S. El-Kady1, S.A. Sidaros 2, S.A.El-Kewey2, Eman A.H. Khatab1, Hala A. Amin1 , A.A. Emeran2, and Samaa Abd El-Khalik3

1 Virus and Phytoplasm Res. Dept. Plant Pathology Res. Inst, Agricultural Research Center (ARC). 2Agricultural Botany Department, Fac. of Agric., Kafrelsheikh Univ. 3Plant Disease Res. Section, Sakha, Kafr El-Sheikh Plant Pathology Research Institute ARC, Egyp.t

Received October 2007: accepted December2007

Abstract

Pepper mild mottle virus was purified from infected (N. clevelandii) plants. The UV-absorption spectrum had a maximum at 260 nm and a minimum at 248 nm. The ratios of Amax/ Amin and A260/ A280 were 1.11 and 1.21, respectively. The yield of purified PMMoV preparation was about 16.04 mg/100g of infected tissue. Electron micrograph of purified virus preparation revealed rod-shaped particles of about 312 nm long and 18 nm width typical of tobamoviruses. Polyclonal antibodies raised against PMMoV has a maximum titer of 1: 1024 from bleeding taken , two weeks after the last injection. The dilution end point of PMMoV in infected tissue extracts was 1/640 as determined by indirect ELISA. Detection of PMMoV in infected plants had been readily applied by DBIA and TBIA on nitrocellulose membranes using the produced antiserum