Document Type : Research Paper

Authors

• davod koolivand ¹
• nemat sokhandan bashir ²
• ali akbar behjat nia ³

¹ Assistant Professor, Department of Plant Protection, Faculty of Agriculture, University of Zanjan

² Associate Professor, Department of Plant Protection, Faculty of Agriculture, University of Zanjan

³ Professor, Plant Virology Research Center, College of Agriculture, Shiraz University, Shiraz, Iran

Abstract

Among several diseases of gapaevine, fan-leaf disese is the most imprtant viral diseases in the garapevine orchards around the world and Iran. Furthermore, early detection and diagnosis of the disease and screning the infected plant could be useful to disase management. In the recent research, local GFLV isolate was selected to prepare the recombinant antibody to diagnosis the infected samples. In the recent research, after the rabbit immunization by coat protein and movement protain of GFLV that expressed into E.coli, the rabbits were bled and the serum fractions were collected and stored at -20°C until required. After the titration of serum fraction, the immunoglobulin G (IgG) from the antiserum was purified using DEAE cellulose column and the efficiency of purification was estimated by the light absorption at 280 nm (A280) and electrophoresis on 12% gel SDS-PAGE. After the IgG purification, indirect-ELISA was done by different dilution including 1:500, 1:1000 and 1:2000. The result showed, IgG-CP-GFLV revelaed the best absorbance in 1:500 and 1:1000 IgG-cp diluted wherase the IgG-MP-GFLV showed the best reaction in 1:500 IgG-MP dilution. DIBA by purified IgGs showed that the IgGs could be detect the related antigenes in 1:500 dilution fot both. Finally, the result revealed that IgGs purified by DEAE cellulose column could be applied in serological and seromolecular test.

Keywords

• Antibody
• Coat Protein
• Movement Protein
• Grapevine Fanleaf Virus