Li Zhuo, Chen Huan, Yuan De Peng, Jiang Xu, Li Zhi Min, Wang Si Ting, Zhou Tian Ge, Zhu Hong Yao, Bian Qiang, Zhu Xiao Feng, Xuan Yuan Hu
College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China.
State Key Laboratory of Elemento-Organic Chemistry and Department of Plant Protection, National Pesticide Engineering Research Center (Tianjin), Nankai University, Tianjin 300071, China.
J Adv Res. 2025 May;71:93-106. doi: 10.1016/j.jare.2024.05.032. Epub 2024 May 31.
Rhizoctonia solani Kühn is a pathogen causing rice sheath blight (ShB). Ammonium transporter 1 (AMT1) promotes resistance of rice to ShB by activating ethylene signaling. However, how AMT1 activates ethylene signaling remains unclear.
In this study, the indeterminate domain 10 (IDD10)-NAC079 interaction model was used to investigate whether ethylene signaling is modulated downstream of ammonium signaling and modulates ammonium-mediated ShB resistance.
RT-qPCR assay was used to identify the relative expression levels of nitrogen and ethylene related genes. Yeast two-hybrid assays, Bimolecular fluorescence complementation (BiFC) and Co-immunoprecipitation (Co-IP) assay were conducted to verify the IDD10-NAC079-calcineurin B-like interacting protein kinase 31 (CIPK31) transcriptional complex. Yeast one-hybrid assay, Chromatin immunoprecipitation (ChIP) assay, and Electrophoretic mobility shift assay (EMSA) were used to verify whether ETR2 was activated by IDD10 and NAC079. Ethylene quantification assay was used to verify ethylene content in IDD10 transgenic plants. Genetic analysis is used to detect the response of IDD10, NAC079 and CIPK31 to ShB infestation.
IDD10-NAC079 forms a transcription complex that activates ETR2 to inhibit the ethylene signaling pathway to negatively regulating ShB resistance. CIPK31 interacts and phosphorylates NAC079 to enhance its transcriptional activation activity. In addition, AMT1-mediated ammonium absorption and subsequent N assimilation inhibit the expression of IDD10 and CIPK31 to activate the ethylene signaling pathway, which positively regulates ShB resistance.
The study identified the link between ammonium and ethylene signaling and improved the understanding of the rice resistance mechanism.
立枯丝核菌是引起水稻纹枯病的病原菌。铵转运蛋白1（AMT1）通过激活乙烯信号通路来促进水稻对纹枯病的抗性。然而，AMT1如何激活乙烯信号通路仍不清楚。
在本研究中，利用不定结构域10（IDD10）-NAC079相互作用模型来研究乙烯信号是否在铵信号下游被调节，并调节铵介导的对纹枯病的抗性。
采用RT-qPCR分析法来鉴定氮和乙烯相关基因的相对表达水平。进行酵母双杂交试验、双分子荧光互补（BiFC）和免疫共沉淀（Co-IP）试验以验证IDD10-NAC079-类钙调神经磷酸酶B互作蛋白激酶31（CIPK31）转录复合体。利用酵母单杂交试验、染色质免疫沉淀（ChIP）试验和电泳迁移率变动分析（EMSA）来验证乙烯受体2（ETR2）是否被IDD10和NAC079激活。采用乙烯定量分析法来验证IDD10转基因植物中的乙烯含量。利用遗传分析来检测IDD10、NAC079和CIPK31对纹枯病侵染的反应。
IDD10-NAC079形成一个转录复合体，该复合体激活ETR2以抑制乙烯信号通路，从而对纹枯病抗性起负调控作用。CIPK31与NAC079相互作用并使其磷酸化，以增强其转录激活活性。此外，AMT1介导的铵吸收及随后的氮同化抑制IDD10和CIPK31的表达，从而激活乙烯信号通路，对纹枯病抗性起正调控作用。
该研究确定了铵信号与乙烯信号之间的联系，增进了对水稻抗性机制的理解。
