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干旱胁迫下平邑甜茶叶片差异表达蛋白分析
杨小妮1, 王俊花1, 杨剑锋,等1
西北农林科技大学 园艺学院
摘要:
【目的】探究平邑甜茶在干旱胁迫下的蛋白应激反应,为苹果砧木抗旱分子改良提供参考。【方法】以苹果砧木平邑甜茶为试材,在正常灌水后对其进行干旱胁迫处理(Mh-GD),以持续灌水为对照(Mh-CK),采用生理学和蛋白质组学相结合的研究方法,测定土壤水势、黎明前叶水势、光合作用及其叶片荧光参数,运用iTRAQ技术筛选鉴定响应干旱胁迫的差异表达蛋白,并对成功鉴定的蛋白进行GO 注释及参与 KEGG 通路的分析。【结果】①与Mh-CK处理相比,Mh-GD处理平邑甜茶叶片相对含水量(RWC)、黎明前叶水势(ψleaf)、光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)均显著下降。②Mh-GD处理植株叶片PSⅡ的最大光合效率(Fv/Fm)、潜在活性(Fv/F0)、性能指标(PI)、单位反应中心吸收的光能(ABS/RC)均降低,而荧光动力学曲线升高。③运用iTRAQ技术共鉴定出1 897个蛋白,其中181个蛋白质表达量在Mh-CK、Mh-GD处理叶片间有显著差异,上调蛋白81个(P<0.05,差异倍数>1.5),下调蛋白100个(P<0.05,差异倍数<0.67)。181个蛋白中,86个蛋白功能已知,95个蛋白功能未知;已知功能可归为10类。【结论】在干旱胁迫下,平邑甜茶的差异表达蛋白质主要参与了光合作用、抗逆、信号转导和碳代谢等过程。
关键词:  苹果砧木  蛋白组  叶片  水分逆境
DOI:
分类号:
基金项目:国家苹果产业技术体系项目(CARS-27);陕西省科技统筹创新工程计划项目(2015KJZDNY02-03-02)
Proteomic analysis of apple leaf under water stress
YANG Xiaoni,WANG Junhua,YANG Jianfeng,et al
Abstract:
【Objective】The proteomic response of Malus hupehensis (Mh) involved in water deficit stress was evaluated to provide basis for improving apple rootstock drought resistant breeding.【Method】The apple rootstock of Malus hupehensis was used for test with the control (Mh-CK) of continuous irrigation and gradual drought (Mh-GD) treatment of persistent drought after irrigation.The combined methods of physiology and proteomics were adopted to measure soil and leaf water potential,photosynthesis and chlorophyll fluorescence parameters.The iTRAQ technology was used to identify the differentially expressed proteins in Malus hupehensis leaf under water stress.The proteins were also analyzed by GO annotation and KEGG pathway analysis.【Result】① Leaf relative water content (RWC),predawn leaf potential (ψleaf),photosynthesis (Pn),transpiration rate (Tr) and stomatal conductance (Gs) of Mh-GD plants were decreased significantly compared with Mh-CK.② Chlorophyll fluorescence parameters such as PSⅡ the maximum photosynthetic efficiency (Fv/Fm),PSII potential activity (Fv/F0),performance index (PI),absorption flux per RC (ABS/RC) of Mh-GD were lower than those of Mh-CK,while the chlorophyll fluorescence intensity was higher.③ A total of 1 897 proteins were identified by iTRAQ,of which 181 were significantly differently expressed in Mh-CK and Mh-GD leaves.Among the 181 proteins, 81 were up-regulated (P<0.05,change fold>1.5) and 100 were down regulated (P<0.05,change fold<0.67).Among the 181 DEPs,86 functions are known,and 95 functions are unknown.Known functions can be classified into 10 categories.【Conclusion】Under gradual drought stress,the Mh proteins responding to water stress mainly involved in photosynthesis pathway,resistance,signal regulated and carbon metabolism.
Key words:  apple rootstock  proteome  leaf  water deficit stress