Partial charaterization of an isolate of maize dwarf mosaic potyvirus and production of polyclonal antibodies for virus detection

摘要

Maize and sorghum crops are subjected to several virus diseases. The virus disease, in particularly, maize dwarf mosaic potyvirus (MDMV) was considered as one of the most important viruses which cause considerable losses in yield productivity as well as the quality of the crop, respectively. The present investigation was conducted to study the characterization of a MDMV isolate based on the biological, molecular and serological tools. Mosaic and severe mosaic symptoms were observed on Zea mays cv.Pioneer and Sorghum bicolor cv. Rio; respectively, which were mechanically inoculated with MDMV-isolate. The stability of this isolate was recognized as TIP, DEP and LIV and found to be 55 deg C. 1/500 and 24 hr, respectively. The electron microscopy ofultrathin sections of virus-infected maize leaf showed that cylindrical inclusion bodies as pin-wheels, scrolls and laminated aggregates were reported in the cytoplasm of MDMV-infected maize leaves. MDMV was purified based on sucrose density gradient centrifugation with a yield of about 2.43 mg/lOOg MDMV-infected maize leaves. MDMV particles were reported to be flexuous filamentous ranging in length of 725-750 nm and about 13 nm in width and contain a coat protein of about 39 KDa. Polyclonal antibodies(PAbs) against the MDMV isolate were raised in rabbits and the antiserum liter was determined using direct antigen coating-enzyme-linked immunosorbent assay (DAC-ELISA). MDMV-antiserum liter was 1/1000, 1/2000 and 1/1500 for first, second and third bleeding, respectively. These PAbs could be useful for preparing an ELISA kit for virus detection.