非水力根源信号调控下冬小麦苗期叶片差异表达蛋白分析

Differential Proteomics Analysis in Winter Wheat Seedling Stage under Non-hydraulic Root-sourced Signals Regulation

从分析干旱胁迫下冬小麦苗期非水力根源信号(n HRS)的产生及其调控下差异表达蛋白的特征和功能,初步揭示小麦抗旱调节机制。本研究以冬小麦‘长武131’为研究材料,通过盆栽控制实验,研究逐渐干旱(12 d)过程中冬小麦苗期生理指标和蛋白质水平的变化。结果显示,干旱胁迫促进小麦叶片ABA积累,诱导气孔关闭,激发nHRS的产生,降低叶片相对含水量。利用串联质谱标记(TMT)蛋白质组学技术对nHRS作用阶段的叶片进行分析,发现干旱胁迫处理与对照相比,共有93个显著差异表达蛋白,其中上调55个(p<0.05,差异倍数>1.2),下调38个(p<0.05,差异倍数<0.83)。对差异表达蛋白进行基因本体论(GO)注释,差异表达蛋白主要参与代谢过程、生物合成过程和生物调控等重要生物学过程,主要集中分布在细胞质中,具有结合、催化活性和抗氧化活性等分子功能。京都基因与基因组百科全书(KEGG)通路注释表明,两种处理的差异表达蛋白富集于48个代谢通路中,主要位于内质网蛋白加工、氨基酸生物合成、二羧酸代谢、碳代谢、光合作用、氮代谢、氧化磷酸化等代谢通路。将差异表达蛋白归类到调节蛋白(7.5%)、代谢和能量(6.5%)、细胞骨架(1.1%)、其他功能(2.2%)和功能未知(81.7%) 5个蛋白功能分类中。从功能分类看,差异蛋白主要参与了干旱胁迫应答、代谢和能量、气孔运动和细胞骨架等过程。本研究为抗旱性小麦的育种和小麦高产节水耕作技术提供了理论依据。

In order to analyze the generation of nHRS(non-hydraulic root-sourced signals)in winter wheat during seedling stage,and the characteristics and functions of differentially expressed proteins under the regulation of nHRS,and preliminarily reveal the mechanism of drought resistance regulation in wheat.In this study,'Changwu 131'was used as the experimental material to study the changes of physiological indexes and protein level of winter wheat seedling during the gradual drought stress(12 d).The results showed that drought stress promoted leaf ABA accumulation,induced stomatal closure,stimulated nHRS production and reduced leaf relative water content.TMT proteomic analysis found that 93 significant differentially expressed proteins(DEPs)were induced under the regulation of the nHRS,of which 55 were up-regulated and 38 were down regulated.GO annotation showed that the DEPs were mainly involved in the metabolic process,biosynthesis process,biological regulation and other important biological processes,mainly distributed in the cytoplasm,with binding,catalytic activity,antioxidant activity and other molecular functions.KEGG pathway annotation showed that the differentially expressed proteins were enriched in 48 metabolic pathways,mainly located in endoplasmic reticulum protein processing,amino acid biosynthesis,dicarboxylic acid metabolism,carbon metabolism,photosynthesis,nitrogen metabolism,oxidative phosphorylation and other metabolic pathways.According to the DEPs involved metabolic pathways and biochemical functions,they were classified into five categories,mainly involved in stress defense,metabolism and energy,stomatal movement,and cytoskeleton.This study provided a theoretical basis for the breeding of drought-resistant wheat and wheat high-yield and water-saving tillage technology.