干旱胁迫下平邑甜茶叶片差异表达蛋白分析  

Proteomic analysis of apple leaf under water stress

作　　者：杨小妮[1] 王俊花 杨剑锋 祁香宁 段宇敏 韩明玉[1] 张林森[1] YANG Xiaoni;WANG Junhua;YANG Jianfeng;QI Xiangning;DUAN Yumin;HAN Mingyu;ZHANG Linsen(College of Horticulture,Northwest A&F University,Yangling,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学园艺学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2018年第11期70-80,共11页Journal of Northwest A&F University(Natural Science Edition)

基　　金：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)

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MhCK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。(2)Mh-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。【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】①Leaf 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.②Chlorophyll fluorescence parameters such as PSⅡthe maximum photosynthetic efficiency(F v/F m),PSII potential activity(F v/F 0),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.③A 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.

关 键 词：苹果砧木 蛋白组 叶片 水分逆境 

分 类 号：S661.1[农业科学—果树学]