Tomato brown rugose fruit virus associated with leaf mosaic, mottling and brown rugose patches on fruits of tomato in India

Eleven fruit samples from the tomato plants exhibiting different kinds of symptoms on leaves and fruits were collected from farmers fields located in two states (Karnataka and Maharashtra) of India. To confirm the identity of the pathogen, eleven tomato fruit samples were subjected DAS-ELISA, which revealed that all samples showed positive reaction to ToBRFV antibodies. Further, total RNA was isolated from infected fruits and subjected to RT-PCR assay using tomato brown rugose fruit virus (ToBRFV) coat protein gene specific primers. The resulting PCR amplicons of ~ 500 bp from eleven tomato samples were cloned, and sequenced. Sequence analysis of the CP gene from eleven isolates showed nucleotide identity of 100% with several ToBRFV isolates reported from different countries around the world. This result was further confirmed through phylogenetic analysis, wherein the eleven ToBRFV isolates are clustered closely with isolates reported from different parts of the world. To the best of our knowledge, this is the first record of ToBRFV infecting tomato in India.