Organoarsenic arsanilic acid (ASA) contamination of paddy soil is a serious but less concerned hazard to agriculture and health of people consuming rice as staple food, for rice is one major pathway of arsenic (As) exposure to human food. To date little research has studied the effect of ASA on biochemical process of rice. Silicon (Si) application is able to reduce the toxicities of heavy metals in numerous plants, but little information about ASA. This work investigated whether and how Si influenced alleviation of ASA toxicity in rice at biochemical level to have a better understanding of defense mechanism by Si against ASA stress. Results showed that ASA reduced rice growth, disturbed protein metabolism, increased lipid peroxidation but decreased the efficiencies of antioxidant activities compared to control plants, more severe in roots than in shoots. The addition of Si in ASA-stressed rice plants noticeably increased growth and development as well as soluble protein contents, but decreased malondialdehyde (MDA) contents in ASA-stressed rice plants, suggesting that Si did have critical roles in ASA detoxification in rice. Furthermore, increased superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities along with elevated glutathione (GSH) and ascorbic acid (AsA) contents implied the active involvement of ROS scavenging and played, at least in part, to Si-mediated alleviation of ASA toxicity in rice, and these changes were related to rice genotypes and tissues. The study provided physio-chemical mechanistic evidence on the beneficial effect of Si on organoarsenic ASA toxicity in rice seedlings.