Identification of Sugarcane Mosaic Potyvirus Strains in Egypt
Identification of Sugarcane Mosaic Potyvirus Strains in Egypt
A.l. Abd El-Fattah; A.s. Saclik M.M. El-Kholi; I.A. Åbdcl-Hmnid and M.A. Madkour
ABSTRACT
Sugarcane mosaic pot.1Virus (SCMV) is the causal agent Of mosaic disease which is reported to be one of the most important viral diseases attacking sugarcane crop in Egypt. Therefore. the present work was designed to screen the SCMV strains in Egypt based on biological. serological and molecular characterization. Data of the use of Sorghum bicolor vars. Rio and Atlas show successful detection of 7 SCMV strains (A, B. D. E. H. l. and M). The indirect-enzyme-linked immunosorbent assay (I-ELISA) detection Of SCMV strains showed the presence or five strains. i.e., SCMV-A. SCMV-B. SCMV-D, SrMV-H and SrMV-I. The electron microscopy of SCMV-E-infected sorghum plant cells proved the presence or the cytoplasmic inclusions characterized to the potyviruses. The virus was purified and an antiserum containing polyclonal antibodies specific to SCMVE strain was raised. Western blot immunoassay (WBIA) successfully used to study the specificity of the produced antiserum to SCMV-E strain. On the level of the molecular studies of SCMV•E strain. a single protein band "ith a size of about 35-37 KDa was obtained after SDS-PAGE analysis of the purified virus preparation. The SCMV-E-RNA was isolated from virus-infected sugarcane tissues and then used as a template for reverse used transcription-polymerase to amplify the coat protein chain reaction (CP) gene (RT-PCR) Of SCMV-E to amplify strain. the ThecDNA that directly amplified cp gene was used as a template using the internal primer combination in PCR for confirmation its specificity to the SCMV-cp gene as a PCR product with a size Of about 400 bp was amplified. The cp gene PCR product was cloned into the pGEM-T easy vector and introduced into Escherichia coli strain DH5_ and the plasmid was then isolated. purified for sequencing the nested PCR product. The similarity between the present sequence (SCMVEgvl) and that or the 14 overseas strains belonging to dillërent SCMV groups " as determined.
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December 2004
Vol.1, Iss. 1, Pages 1-564
