National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
Mol Cell Proteomics. 2013 Dec;12(12):3690-703. doi: 10.1074/mcp.M113.031013. Epub 2013 Sep 9.
Verticillium wilt causes massive annual losses of cotton yield, but the mechanism of cotton resistance to Verticillium dahliae is complex and poorly understood. In this study, a comparative proteomic analysis was performed in resistant cotton (Gossypium barbadense cv7124) on infection with V. dahliae. A total of 188 differentially expressed proteins were identified by mass spectrometry (MALDI-TOF/TOF) analysis and could be classified into 17 biological processes based on Gene Ontology annotation. Most of these proteins were implicated in stimulus response, cellular processes and metabolic processes. Based on the proteomic analysis, several genes involved in secondary metabolism, reactive oxygen burst and phytohormone signaling pathways were identified for further physiological and molecular analysis. The roles of the corresponding genes were further characterized by employing virus-induced gene silencing (VIGS). Based on the results, we suggest that the production of gossypol is sufficient to affect the cotton resistance to V. dahliae. Silencing of GbCAD1, a key enzyme involving in gossypol biosynthesis, compromised cotton resistance to V. dahliae. Reactive oxygen species and salicylic acid signaling may be also implicated as regulators in cotton responsive to V. dahliae according to the analysis of GbSSI2, an important regulator in the crosstalk between salicylic acid and jasmonic acid signal pathways. Moreover, brassinosteroids and jasmonic acid signaling may play essential roles in the cotton disease resistance to V. dahliae. The brassinosteroids signaling was activated in cotton on inoculation with V. dahliae and the disease resistance of cotton was enhanced after exogenous application of brassinolide. Meanwhile, jasmonic acid signaling was also activated in cotton after inoculation with V. dahliae and brassinolide application. These data provide highlights in the molecular basis of cotton resistance to V. dahliae.
黄萎病导致棉花产量每年大量损失，但棉花对黄萎病菌的抗性机制复杂，目前仍不清楚。本研究采用比较蛋白质组学方法，分析了感病棉花（Gossypium barbadense cv7124）接种黄萎病菌后的蛋白质组变化。通过基质辅助激光解吸电离飞行时间/飞行时间（MALDI-TOF/TOF）质谱分析，共鉴定到 188 个差异表达蛋白，根据基因本体（GO）注释，这些蛋白可分为 17 个生物学过程。这些蛋白大多数与刺激反应、细胞过程和代谢过程有关。基于蛋白质组学分析，鉴定到了几个参与次生代谢、活性氧爆发和植物激素信号通路的基因，并进一步进行了生理和分子分析。利用病毒诱导的基因沉默（VIGS）技术，进一步研究了这些基因的功能。根据结果，我们认为棉酚的产生足以影响棉花对黄萎病菌的抗性。关键酶 GbCAD1 基因沉默后，棉花对黄萎病菌的抗性降低。根据 GbSSI2 分析，活性氧和水杨酸信号可能也参与了棉花对黄萎病菌的响应调控，GbSSI2 是水杨酸和茉莉酸信号通路交叉的一个重要调节因子。此外，油菜素内酯和茉莉酸信号可能在棉花对黄萎病菌的抗性中发挥重要作用。接种黄萎病菌后，油菜素内酯激活了棉花的油菜素内酯信号通路，外源施加油菜素内酯增强了棉花的抗病性。同时，接种黄萎病菌后，茉莉酸信号也被激活，外源施加油菜素内酯后该信号进一步增强。这些数据为棉花对黄萎病菌抗性的分子基础提供了重要线索。
