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The present study analysed time (0.5 h to 24 h) and tissue [roots, old leaves (OL) and young leaves (YL)] dependent nature of arsenic (As) accumulation and ensuing responses in two contrasting varieties of rice (Oryza sativa L.); Pooja (tolerant) and CO-50 (moderately sensitive). Arsenic accumulation was 5.4-, 4.7- and 7.3-fold higher at 24 h in roots, OL and YL, respectively of var. CO-50 than that in var. Pooja. Arsenic accumulation in YL depicted a delayed accumulation; at 2 h onwards in var. Pooja (0.23 µg g−1 dw) while at 1 h onwards in var. CO50 (0.26 µg g−1 dw). The responses of oxidative stress parameters, antioxidant enzymes, metabolites and ions were also found to be tissue- and time-dependent and depicted differential pattern in the two varieties. Among hormone, salicylic acid and abscisic acid showed variable response in var. Pooja and var. CO-50. Metabolite analysis depicted an involvement of various metabolites in As stress responses of two varieties. In conclusion, an early sensing of the As stress, proper coordination of hormones, biochemical responses, ionic and metabolic profiles allowed var. Pooja to resist As stress and reduce As accumulation more effectively as compared to that of var. CO-50. ga1 • T T Tracking As stress responses of contrasting tolerant (Pooja) and moderately sensitive (CO-50) rice varieties. • Arsenic accumulation was higher in CO-50 than in Pooja in root, OL and YL. • Root, OL and YL showed contrasting responses for hormones (SA, ABA, JA) and ions (K+, Ca2+). • Metabolite profiling showed significant up-regulation of phenolics and flavonoids in Pooja. [ABSTRACT FROM AUTHOR]