College of Life Sciences, National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, 210095, China; Shandong Key Laboratory of Biophysics, Institute of Biophysics, Dezhou University, Dezhou, 253023, China.
College of Life Sciences, National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, 210095, China.
Plant Physiol Biochem. 2024 Feb;207:108423. doi: 10.1016/j.plaphy.2024.108423. Epub 2024 Feb 11.
Salinity is a significant abiotic stress factor affecting plant growth, consequently reducing crop yield. Abscisic acid (ABA), a well-known phytohormone, is crucial in conferring resistance to abiotic stress, thus, understanding the mechanisms underlying ABA biosynthesis is crucial. In rice (Oryza sativa L.), OsABA2, a short-chain dehydrogenase protein, has a pivotal role in modulating ABA biosynthesis and salt tolerance by undergoing phosphorylation at Ser197 through mitogen-activated protein kinase OsMPK1. However, the interaction between OsABA2 and other proteins in regulating ABA biosynthesis remains unclear. We employed OsABA2 as a bait in yeast two-hybrid screening: a basic helix-loop-helix transcription factor interacting with OsABA2, named OsbHLH110, was identified. Our results showed that firefly luciferase complementary imaging, pull-down, and co-immunoprecipitation assays validated the interaction between OsbHLH110 and OsABA2, affirming their interaction in vivo and in vitro. Moreover, the expression of OsbHLH110 significantly increases in response to salt and ABA treatments. Additionally, OsbHLH110 can directly bind to the G-box element in the OsABA2 promoter. This binding enhances luciferase activity controlled by the OsABA2 promoter, thereby increasing the expression of the OsABA2 gene and content of the OsABA2 protein, resulting in an increase in ABA content. OsABA2 enhanced the interaction between OsbHLH110 and OsABA2 promoter. This collaborative effect enhanced the regulation of ABA biosynthesis. Subsequent genetic analysis demonstrated that OsbHLH110 improved the tolerance of rice to salt stress.
盐度是影响植物生长的重要非生物胁迫因素,从而降低作物产量。脱落酸(ABA)作为一种知名的植物激素,在赋予植物对非生物胁迫的抗性方面起着关键作用,因此,了解 ABA 生物合成的机制至关重要。在水稻(Oryza sativa L.)中,短链脱氢酶蛋白 OsABA2 通过丝裂原活化蛋白激酶 OsMPK1 使 Ser197 发生磷酸化,从而在调节 ABA 生物合成和耐盐性方面发挥着重要作用。然而,OsABA2 与其他调节 ABA 生物合成的蛋白质之间的相互作用仍不清楚。我们以 OsABA2 为诱饵进行酵母双杂交筛选,鉴定到与 OsABA2 相互作用的碱性螺旋-环-螺旋转录因子 OsbHLH110。结果表明,萤火虫荧光素酶互补成像、Pull-down 和 co-immunoprecipitation 实验验证了 OsbHLH110 与 OsABA2 之间的相互作用,证实了它们在体内和体外的相互作用。此外,OsbHLH110 的表达在盐和 ABA 处理后显著增加。此外,OsbHLH110 可以直接结合到 OsABA2 启动子中的 G-Box 元件。这种结合增强了由 OsABA2 启动子控制的荧光素酶活性,从而增加了 OsABA2 基因的表达和 OsABA2 蛋白的含量,导致 ABA 含量增加。OsABA2 增强了 OsbHLH110 与 OsABA2 启动子之间的相互作用。这种协同作用增强了对 ABA 生物合成的调控。随后的遗传分析表明,OsbHLH110 提高了水稻对盐胁迫的耐受性。