Key Laboratory of Agricultural Biological Functional Genes, Northeast Agricultural University, Harbin 150030, China.
School of Life Science, Jiangsu Normal University, Xuzhou 221116, China.
Int J Mol Sci. 2023 Jan 8;24(2):1225. doi: 10.3390/ijms24021225.
SnRK1 protein kinase plays hub roles in plant carbon and nitrogen metabolism. However, the function of SnRK1 in legume nodulation and symbiotic nitrogen fixation is still elusive. In this study, we identified GmNodH, a putative sulfotransferase, as an interacting protein of GmSnRK1 by yeast two-hybrid screen. The qRT-PCR assays indicate that gene is highly expressed in soybean roots and could be induced by rhizobial infection and nitrate stress. Fluorescence microscopic analyses showed that GmNodH was colocalized with GsSnRK1 on plasma membrane. The physical interaction between GmNodH and GmSnRK1 was further verified by using split-luciferase complementary assay and pull-down approaches. In vitro phosphorylation assay showed that GmSnRK1 could phosphorylate GmNodH at Ser193. To dissect the function and genetic relationship of GmSnRK1 and GmNodH in soybean, we co-expressed the wild-type and mutated and genes in soybean hairy roots and found that co-expression of / genes significantly promoted soybean nodulation rates and the expression levels of nodulation-related and genes. Taken together, this study provides the first biological evidence that GmSnRK1 may interact with and phosphorylate GmNodH to synergistically regulate soybean nodulation.
SnRK1 蛋白激酶在植物碳氮代谢中起着核心作用。然而,SnRK1 在豆科植物结瘤和共生固氮中的功能仍然难以捉摸。在这项研究中,我们通过酵母双杂交筛选鉴定出 GmNodH,一种假定的磺基转移酶,是 GmSnRK1 的互作蛋白。qRT-PCR 分析表明,该基因在大豆根中高度表达,可被根瘤菌感染和硝酸盐胁迫诱导。荧光显微镜分析表明,GmNodH 与 GsSnRK1 共定位于质膜上。通过使用分裂萤光素酶互补测定和下拉方法进一步验证了 GmNodH 和 GmSnRK1 之间的物理相互作用。体外磷酸化实验表明 GmSnRK1 可在 Ser193 处磷酸化 GmNodH。为了剖析 GmSnRK1 和 GmNodH 在大豆中的功能和遗传关系,我们在大豆毛状根中共表达野生型和突变型 和 基因,并发现 / 基因的共表达显著促进了大豆的结瘤率以及结瘤相关基因 和 的表达水平。总之,这项研究提供了第一个生物学证据,表明 GmSnRK1 可能与 GmNodH 相互作用并磷酸化 GmNodH,以协同调节大豆结瘤。