Iron toxicity is an abiotic stress comes with high concentrations of Fe2+ in the soil solution which is a well-recognized problem of rice (Oryza sativa L.) cultivation in lowland. Rice varieties differ widely in their ability to tolerate excess iron. The present study was undertaken with four rice varieties viz. Dhruba, Sampriti, Dhiren and Puspa. The objective is to study the influence of high applied Fe2+ concentrations on the growth, chlorophyll content, and antioxidant enzyme activities. The spectral reflectivity and absorption of different chemical bonding through Fourier-transform infrared spectroscopy (FTIR) of four rice varieties was also analyzed. The seven days old rice seedlings were treated with Ferrous sulphate subjected to 100 ppm to 750 ppm for further 14 days iron stress was used to analyse the morphological and biochemical responses. Besides, Fourier transform infrared spectral reflection was attributed in root and shoot part. The results indicated shoot growth and chlorophyll content decreased in 750 ppm in all the selected rice varieties of interest. On the contrary the catalase activity, protein content and lipid peroxidation increased in these varieties. However, the expression of high amount of CAT activity in Sampriti variety and high amount of SOD activity in Dhruba variety leads to tolerance in iron stress in comparison to other two varieties of interest. FTIR revealed steep band stretching of various functional groups of different compounds in both the root and shoot part of all the varieties. Manuscript profile