Overexpression of OsPIN5b Alters Plant Architecture and Impairs Cold Tolerance in Rice (Oryza sativa L.)

Auxin plays a versatile role in regulating plant growth and development. The auxin efflux carrier PIN-FORMED (PIN) proteins dictate the distribution and maximum of auxin within various tissues. Despite extensive research on OsPINs in recent years, their functions in abiotic stress resistance, particularly cold tolerance, remain poorly understood. Here, we investigated the role of OsPIN5b in rice (Oryza sativa L.) growth and development, as well as its contribution to cold tolerance using overexpression technology. Overexpression of OsPIN5b (OE) resulted in reduced shoot height and a lower number of adventitious roots at the seedling stage. Transgenic rice plants exhibited an earlier heading date, stunted growth, and compromised agronomic traits, including shortened panicle length, decreased grain number per panicle, reduced seed size, and lower seed setting rate during the reproductive stage. Auxin content in the transgenic lines was significantly elevated, as indicated by the upregulation of the auxin-responsive gene OsIAA20 and increased auxin levels quantified using a newly developed method. Compared with wild-type plants, the cold tolerance of OE plants was markedly reduced, as evidenced by lower survival rates, higher levels of electrolyte leakage, and increased malondialdehyde (MDA) production following cold treatment. In line with this, the transgenic lines produced less soluble sugar and proline, while accumulating more hydrogen peroxide (H₂O₂) and superoxide anion radicals (O2-) after cold treatment. Furthermore, the activities of antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD), were notably decreased upon cold treatment compared with those in WT plants. Additionally, OsRBOHH, which plays a role in ROS production, was significantly upregulated in transgenic lines both before and after chilling stress, suggesting that OsRBOHH plays a potential role in regulating ROS production. Collectively, overexpression of OsPIN5b substantially disturbs auxin homeostasis, resulting in impaired plant architecture and agronomic traits. More importantly, the upregulation of OsPIN5b compromises rice cold tolerance by perturbing ROS homeostasis and adversely influencing the accumulation of soluble sugar and proline.