Identification of redox-regulated components of arsenate (AsV) tolerance through thiourea supplementation in rice

Abstract

Arsenic (As) is a ubiquitously present environmental carcinogen that enters into the human food chain through rice grains. In our previous research, the application of thiourea (TU; a non-physiological thiol based ROS scavenger) has been demonstrated to enhance salt and UV stress tolerance as well as the crop yield under field conditions. These effects were associated with the ability of TU to maintain plant redox homeostasis. Since As stress also induces redox imbalance, the present research was initiated to evaluate the efficiency of TU in regulating As tolerance/accumulation in rice. The supplementation of TU (75 μM) to As^V (25 μM) improved the root growth and also reduced the As concentration by 56% in the aerial parts, which could be attributed to significant downregulation of the Lsi2 transporter responsible for the translocation of As from root to shoot. The fact that these effects were not due to direct interaction between As and TU was confirmed from complexation studies using HPLC-(ICP-MS)-(ESI-MS). Short-term kinetic studies of GSH levels and the GSH/GSSG ratio confirmed the establishment of differential redox states in As and As + TU treated seedlings. The real-time RT-PCR based comparative expression profiling under As with/without TU treatment identified Sultr1;1 and Sultr1;2 as major redox-regulated sulfate transporters. Their specific induction in shoots coupled with enhanced root-to-shoot sulfate translocation (analyzed using ³⁵S-sulfate as a radiotracer) was observed under TU supplementation. Furthermore, the level of thiolic metabolites (PC2 in roots and GSH and PC3 in shoots) and activities of sulfur metabolism enzymes (ATP sulfurylase and cysteine synthase in roots and 5′-adenylylsulfate reductase in shoot) were also increased with As + TU as compared to As treatment. Thus, this study utilizes the interaction between As and TU to identify the critical redox regulated components of As tolerance in rice.