Liu Shifeng, Cai Chengcheng, Li Luopin, Wen He, Liu Jie, Li Liqin, Wang Qiang, Wang Xiyao
State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130, China.
Hortic Res. 2023 Nov 8;10(12):uhad228. doi: 10.1093/hr/uhad228. eCollection 2023 Dec.
After harvest, potato tubers undergo an important period of dormancy, which significantly impacts potato quality and seed vigor. has been reported as a key gene for maintaining tuber dormancy; in this study, we explored the molecular mechanism by which maintains dormancy. StBIN2 was first identified as a candidate protein that interacts with StSN2 by co-immunoprecipitation/mass spectrometry, and both qPCR and enzyme activity experiments showed that can promote the StBIN2 expression and activity. In addition, the interaction between StSN2 and StBIN2 was verified by yeast two-hybrid, luciferase complementation experiments and co-immunoprecipitation. Bioinformatics analysis and site-directed mutagenesis confirmed the critical role of cysteine residues of StBIN2 in its binding to StSN2. Similar to that of , overexpression of extended the dormancy of potato tuber. Interaction between StSN2 and StBIN2 increased the activity of the StBIN2 enzyme, inhibited the expression of , and suppressed BR signaling. On the contrary, this interaction promoted the expression of // and , key genes of ABA signaling, and the phosphorylation of , thereby promoting ABA signaling. Altogether, our results indicate that StSN2 interacts with StBIN2 through key cysteine residues and StBIN2 maintains tuber dormancy by affecting ABA and BR signaling. Findings of this research offer new insights into the molecular mechanism by which StSN2 maintains potato tuber dormancy through interaction with StSIN2 and provide guidance for potato improvement.
收获后,马铃薯块茎会经历一个重要的休眠期,这对马铃薯品质和种子活力有显著影响。据报道,[某基因]是维持块茎休眠的关键基因;在本研究中,我们探究了[该基因]维持休眠的分子机制。首先通过免疫共沉淀/质谱法鉴定出StBIN2是一种与StSN2相互作用的候选蛋白,定量PCR和酶活性实验均表明[某基因]可促进StBIN2的表达和活性。此外,通过酵母双杂交、荧光素酶互补实验和免疫共沉淀验证了StSN2与StBIN2之间的相互作用。生物信息学分析和定点诱变证实了StBIN2的半胱氨酸残基在其与StSN2结合中的关键作用。与[某基因]类似,[另一基因]的过表达延长了马铃薯块茎的休眠期。StSN2与StBIN2之间的相互作用增加了StBIN2酶的活性,抑制了[某基因]的表达,并抑制了油菜素内酯(BR)信号传导。相反,这种相互作用促进了脱落酸(ABA)信号传导关键基因[相关基因]和[另一相关基因]的表达以及[某蛋白]的磷酸化,从而促进了ABA信号传导。总之,我们的结果表明,StSN2通过关键的半胱氨酸残基与StBIN2相互作用,且StBIN2通过影响ABA和BR信号传导来维持块茎休眠。本研究结果为StSN2通过与StSIN2相互作用维持马铃薯块茎休眠的分子机制提供了新见解,并为马铃薯改良提供了指导。