Abstract: An Egyptian isolate of a Tobamovirus was isolated and identified from Chrysanthemum cultivation in Egypt. Both biological, serological and sequence analysis of the coat protein gene demonstrated that the virus represents an isolate of the Tobamoviridae family. The isolated virus was nominated as TMV Chrysanthemum Egyptian isolate (TMV-Ch-EG). This virus isolate caused severe disease symptoms in Chrysanthemum plants with mosaic, mottling and flower discoloration. The virus was purified biologically using serial transfer of the single local lesion technique on Nicotiana gultinosa. The induced antiserum for the isolated virus had a titer 1\1024. 600 bp DNA fragments from the coat protein gene (CP) of TMV-Ch-EG was amplified with Rt-PCR technique. Phylogenetic analysis of the TMV-Ch-EG/CP- gene showed 89% nucleotide sequence homology with other published strains of TMV in GenBank and 81% amino acid sequence homology. Tissue culture approach was used to permit the recovery of TMV-Ch-free micropropagated shoots via application of 20 mg L-1 virazole followed by thermotherapy at 38°C for two weeks and early screening to facilitate the efficient production of virus-free tissue culture derived propagules using the produced antiserum against TMV-Ch-EG.