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DIAGNOSIS OF MAIZE DWARF MOSAIC POTYVIRUS USING INCLUSION BODIES
DIAGNOSIS OF MAIZE DWARF MOSAIC POTYVIRUS USING INCLUSION BODIES E K ALLAMl,V HARI2,andMH ABDEIGHAFFARl 1Virology Lab, Microbiology Dept, Fac. of Agric., Ain Shams University, Calm, Egypt, 2Biological Scinces Dept, Wayne S University, De MI, 48202, USA. < b> Background and Objectives MDMV-A Is a member of the potyvirus group [1] . Infected plant cells contain characteristic inclusion bodies known as cylindrical inclusions (Cl). The objective of this work was firsty to purify the Cl protein , prepare serum and then confirm the specificty of this antiserum . This antiserum was used for diagnosis of MDMV In infected plants by western blotting and immunogold electron microscopy. Materials and Methods Cylindrical inclusion protein was purified from the same tissue used for virus purification, using a modified procedure based on knv speed centrifugation and rate-zonal sucrose gradients. Molecular weight of MDMV-CI was determined by SDS-PAGE . The protein bands containing the CI were sliced from the gels and used for preparation of Cl antiserum by injecting rabbits sub-cutaneously. TWr of MDMV-CI was estimated by repeatme-linked immunosorbent assay ( R-ELISA) [2]. Electron microscopic and immunogold labelling studies were performed using ultrathin sections of infected leaves. Results and conclusions This work indicated the presence of characteristic inclusion bodies proteins with a MW ~ 70 KDa . Immunogold electron microscopy of tissue sections using the prepared antiserum for MDMV-CI was used to show the presence of scrolls, pinwheels and laminated aggregates inclusions . These results indicated that MDMV was assigned to Subdivision III, according to the types of cylindrical inclusions . The MDMV-CI antiserum can be us as a diagnosis aid , particularly so if it is difficult to obtain antiserum to structural proteins. Andscra to non-structural proteins are also valuable In monitoring the temporal biosesis or accumulation of these proteins in infected plants, cells or protoplasts. Reference 1 - Allam, E k, El-Afifi Sohatr, Abo El-Nasr M and Hussein M (1987. (Abstr.) page 184 in : Abslr. VIII Int Congr. ViroL, Edmenton, Can. 9-14 August NatRes.Counc., Ottawa, 369 pp. 2 - Baunoch, D A, Das p, Browning, M E and Hart V (1992). Biotechniques 12: 412-417.