During characterization of rhizobacteria, strain DBTS2(T) was isolated from the rhizosphere soil samples of healthy tomato plants and characterized using a polyphasic taxonomic approach. Phylogenetic analysis using 16S rRNA gene sequences showed this strain belonged to the genus Rhizobium and was most closely related to Rhizobium subbaraonis JC85(T) (99.1%) and Rhizobium daejeonense CCBAU 10050(T) (97%). Cells of strain DBTS2(T) were Gram-negative, short rod, aerobic and non-motile. This novel strain was found to grow at 20-45 degrees C (optimum 25-37 degrees C), pH 5-9 (optimum 8) and in the presence of 4% NaCl. It was positive for catalase and oxidase. The predominant cellular fatty acids were Summed Feature 8 (52.7%) and C-19:0 cyclo omega 8c (23.3%). The polar lipids of strain DBTS2(T) consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, unidentified aminophospholipid, unidentified aminolipid, four unidentified phospholipids, unidentified lipid, phosphatidylcholine, unknown glycolipid and unknown aminophosphoglycolipids. Q-10 was the major quinone. The DNA-DNA hybridization similarity values between the strain DBTS2(T) and R. subbaraonis JC85(T), R. daejeonense CCBAU 10050(T) and Rhizobium azooxidifex DSM100211(T) were 46.4%, 20.7% and 25.5%, respectively. The ANI value was 91.96% between strain DBTS2(T) and R. subbaraonis JC85(T) and 75.18% between strain DBTS2(T) and R. daejeonense CCBAU 10050(T). The DNA G+C content of the genomic DNA was 63.1 mol%. Based on these results, it was concluded that the isolate represents a novel species of the genus Rhizobium, for which the name Rhizobium rhizolycopersici sp. nov. is proposed, with DBTS2(T) (= CICC 24887(T) = ACCC61707 = JCM 34245) as the type strain.