Rice tonoplast intrinsic protein member OsTIP1;2 confers tolerance to arsenite stress

The International Agency for Research on Cancer categorizes arsenic (As) as a group I carcinogen. Arsenic exposure significantly reduces growth, development, metabolism, and crop yield. Tonoplast intrinsic proteins (TIPs) belong to the major intrinsic protein (MIP) superfamily and transport various substrates, including metals/ metalloids. Our study aimed to characterize rice OsTIP1;2 in As[III] stress response. The gene expression analysis showed that the OsTIP1;2 expression was enhanced in roots on exposure to As[III] treatment. The heterologous expression of OsTIP1;2 in S. cerevisiae mutant lacking YCF1 (ycf1 Delta) complemented the As[III] transport function of the YCF1 transporter but not for boron (B) and arsenate As[V], indicating its substrate selective nature. The ycf1 Delta mutant expressing OsTIP1;2 accumulated more As than the wild type (W303-1A) and ycf1 Delta mutant strain carrying the pYES2.1 vector. OsTIP1;2 activity was partially inhibited in the presence of the aquaporin (AQP) inhibitors. The subcellular localization studies confirmed that OsTIP1;2 is localized to the tonoplast. The transient overexpression of OsTIP1;2 in Nicotiana benthamiana leaves resulted in increased activities of enzymatic and non-enzymatic antioxidants, suggesting a potential role in mitigating oxidative stress induced by As[III]. The transgenic N. tabacum overexpressing OsTIP1;2 displayed an As[III]-tolerant phenotype, with increased fresh