NtPIN3 positively regulates low phosphorus tolerance by changing root elongation, Pi concentration and antioxidant capacity in tobacco

Phosphorus (P) is a critical nutrient for plants, and inorganic orthophosphate (Pi) deficiency results in declining crop quality and yields. Auxin plays a key regulatory effect on plant growth in response to Pi deficiency, yet the regulatory mechanisms remain poorly understood. In this study, we cloned and identified an auxin efflux transporter, NtPIN3, involved in the low phosphate response of tobacco (Nicotiana tabacum). NtPIN3 is located on the cytoplasmic membrane, and high expression levels of NtPIN3 were observed under phosphorus deficiency conditions. Moreover, NtPIN3 overexpression in tobacco could dramatically enhance low Pi tolerance by increasing the root length, which was consistent with the GUS activity and the auxin concentration in the root. Notably, Pi concentration and antioxidant capacity in overexpression transgenic tobacco were significantly increased compared with the wild type (WT) under low Pi stress conditions due to Pi uptake (NtPT1 and NtPT2) and antioxidant capacity (superoxide dismutase, peroxidase, and catalase). In contrast, compared with WT, the ntpin3 mutant significantly reduced root length, auxin concentration, Pi concentration, and antioxidant capacity under low Pi stress conditions, suggesting that NtPIN3 is a positive regulator of low phosphate in tobacco. In addition, the yeast-two-hybrid and luciferase complementation imaging assay confirmed that NtPIN3 interacted with NtPIN2. Altogether, the NtPIN3 involved in low Pi response and overexpression of NtPIN3 contributed to enhanced tolerance to low Pi stress, which proved an underlying gene for breeding low Pi-stress tolerant plants.