iTRAQ技术分析钝叶草抗旱的分子机理

Drought stress is a major factor affects the using and quality of turf grasses. St.Augustingrass was an important turfgrass and widely planted in south china but it was limited by the drought stress. To identify the mechanisms underlying drought stress in St.Augustingrass, drought stress-induced changes in protein profile of St.Augustingrass leaves were comprehensively analyzed using iTRAQ (Isobaric Tag for Relative and Absolute Quantification) differential liquid chromatography-tandem mass spectrometry. The research materials included drought-tolerance mutant 12-6 and drought-senstive CK identified by many years. The different expression proteins were compared from the different drought-tolerance materials. And identify the iTRAQ results with the method of western-blot. Analysis the relationship between the different expression protein and the genes induce by the protein. Make sure the core pathway induced by the different expression protein and built the protein interactions internet in core pathway. And identify protein interactions by bimolecular fluorescence complementation (BiFC). The productions induced by drought stress in core pathway were analyzed by the method of HPLC(high performance liquid chromatography) and GC-MS (chromatography-mass spectrometry). This research was found the molecular mechanism on drought stress from the level of protein, gene and pathway. And the relationships among three levels were verified. It kept the result accuracy and reality. The research was important meaning on improving the germplasm resource and molecular breeding.

干旱胁迫严重影响了草坪草的应用和品质，研究草坪草的抗旱机理尤为重要。蛋白组学iTRAQ技术的应用可以从蛋白角度揭示植物抗逆的分子机理。本研究将以申请者实验室已鉴定的抗性材料突变体12-6钝叶草和敏感材料普通钝叶草（CK）为材料，利用iTRAQ技术分析两种抗性不同的材料响应干旱胁迫的蛋白及蛋白表达的差异，同时利用Western-blot 验证iTRAQ实验结果；对调控差异表达蛋白的基因进行表达分析，分析差异表达蛋白与差异表达基因的关系；明确差异表达蛋白调控的核心代谢途径，构建核心代谢途径中的蛋白互作的网络图谱，应用双分子荧光互补技术验证蛋白互作的关系，并且分析核心代谢途径中的产物响应干旱的情况。本研究是从蛋白水平、基因水平和代谢水平三个方面揭示钝叶草抗旱的分子机理，并且三个方面互相验证，保证研究结果的准确性与真实性。本研究将对钝叶草种质资源的改良与分子遗传育种具有重大意义。