A new strain of Tobacco mosaic virus (TMV) was isolated from soybean (Glycine max) in India (TMV-SoyIn). It produced puckering symptoms on inoculated soybean cultivars, JS-20-30 and JS 93-05, and severe necrotic spots on inoculated leaves of Nicotiana tabacum cv. Xanthi, followed by systemic necrotic spots on subsequent leaves as well as greenish mosaic symptoms on the top leaves. In the electron microscope, tobamovirus like particles were observed in soybean leaves and also in the other inoculated host plants. The host reactions revealed that the present isolate was different from the previously described TMV strain of soybean (TMV-S) from Yugoslavia. To confirm the virus identity, the 3′ end of the genome containing the movement protein gene, coat protein and the 3′ untranslated region of the virus isolate was sequenced (1494 nucleotides; JQ895560). Results indicated that the virus associated with soybean crinkle was an isolate of TMV as the present isolate had 94.4–99.7% sequence identity with known TMV isolates. This is the first report of a TMV-strain infecting soybean in India.