Polyclonal antibodies production against Broad bean mottle virus affecting faba bean (Vicia faba L.) plants in Egypt
Polyclonal antibodies production against Broad bean mottle virus affecting faba bean (Vicia faba L.) plants in Egypt
Eman A. H. Kattab1,3, A. H. Ebrahem2 , Om-Hashem M. El-Banna2 and Hanan F. EL-Kammar³
ABSTRACT
An isolate of BBMV was obtained from naturally infected faba bean plants growing in the experimental fields of Giza Agricultural Research Station showing mottle and distortion of leaves. The identity of the virus was confirmed biologically, serologically and by light and electron microscopy. It was found that BBMV reacted systemically with 12 species belonging to Fabaceae and locally with 2 species belonging to Chenopodiace. It was transmitted mechanically and by Sitona lineate. It was detected serologically by indirect ELISA using BBMV specific antiserum. Light microscopy of epidermal strips of faba bean infected-leaves revealed amorphous cytoplasmic inclusions (X-bodies). The UV absorption spectrum of the purified virus had a maximum at 260 nm and a minimum at 245 nm. The ratios of Amax/Amin and A260/280 were 1.39 and 1.57, respectively. Yield of the purified BBMV preparation was about 2.5 mg/100g of infected tissue. Electron micrograph of purified virus preparation revealed isometric shaped particles of with mean diameter of 26 nm typical of Bromoviruses. Polyclonal antibodies raised against BBMV have maximum titer of 1:1024 from the first bleeding. IgG and IgG conjugate were 1.0 μg/ ml and 1:1000, respectively. The produced antiserum was evaluated by DBIA and TBIA using infected plant.
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