Macrotyloma uniflorum (horse gram) is considered an under-utilized legume crop despite its nutritional and medicinal values. In India, it has wide acceptance among farming communities. This investigation emphasized on the possible application of two endosymbionts (Bacillus sp. AS03 and Rhizobium sp. AS05) of horse gram cultivated on Cr (VI)-contaminated soil. The photosynthetic performance (PIφ) of Cr treated plants co-inoculated with AS03 and AS05 was significantly improved compared with non-inoculated Cr treated plants based on photosynthetic yield, which was evidenced from the rise in the fluorescence at I–P transient and rate of photosynthesis (pN), indicating synergistic action between plant and bacteria (AS03 and AS05). The smooth electron transport from PS II to PS I was achieved in the Cr stressed plants inoculated with both the bacterial strains. The detrimental effects of Cr toxicity on the root tips were also minimized with bioinoculation as revealed from mitotic index. Plants with dual inoculation of AS03 and AS05 had significantly lesser chromosomal aberration in the roots. Dual inoculation biochar or seed inoculation have beneficial impact on the plant photosynthetic performance along with improved growth of roots in plants treated with Cr (VI). The results of the current work suggest the possitive effect of dual inoculation of Cr tolerant endosymbionts, Bacillus sp. (AS03) and nodulating Rhizobium sp. (AS05), in reducing cytological as well as physiological stress of plants in Cr (VI) contaminated soil. Display omitted •Endosymbiotic Bacteria drops Abnormality Index in Macrotyloma under Cr (IV) stress.•Dual inoculation increased photosynthetic performance of Macrotyloma.•Chlorophyll fluorescence parameters showed synergistic effect of dual inoculation.