Dynamic responses ofPAto environmental stimuli imaged by a genetically encoded mobilizable fluorescent sensor  

作　　者：Teng Li Xingkai Xiao Qingyun Liu Wenyan Li Li Li Wenhua Zhang Teun Munnik Xuemin Wang Qun Zhang 

机构地区：[1]College of Life Sciences,State Key Laboratory of Crop Genetics and Germplasm Enhancement,Nanjing Agricultural University,Nanjing 210095,China [2]Institute of Botany,Jiangsu Province and Chinese Academy of Sciences(Nanjing Botanical Garden Mem.Sun Yat-Sen),Nanjing 210014,China [3]Cluster Green Life Sciences,Section Plant Cell Biology,Swammerdam Institute for Life Sciences,University of Amsterdam,Amsterdam,the Netherlands [4]Department of Biology,University of Missouri-St.Louis,St.Louis,MO 63121,USA [5]Donald Danforth Plant Science Center,St.Louis,MO 63132,USA

出　　处：《Plant Communications》2023年第3期99-111,共13页植物通讯（英文）

基　　金：supported by the National Key R&D Program of China(2022YFA1303400 and 2022YFD1201700);the National Natural Science Foundation of China(32270301 and 31970300);the Jiangsu Agriculture Science and Technology Innovation Fund(CX-20-2007);the Fundamental Research Funds for the Central Universities(XUEKEN2022002).

摘　　要：Membrane fluidity,permeability,and surface charges are controlled by phospholipid metabolism and transport.Despite the importance of phosphatidic acid(PA)as a bioactive molecule,the mechanical properties of PA translocation and subcellular accumulation are unknown.Here,we used a mobilizable,highly responsive genetically encoded fluorescent indicator,green fluorescent protein(GFP)-N160RbohD,to monitor PA dynamics in living cells.The majority of GFP-N160RbohD accumulated at the plasma membrane and sensitively responded to changes in PA levels.Cellular,pharmacological,and genetic analyses illustrated that both salinity and abscisic acid rapidly enhanced GFP-N160RbohD fluorescence at the plasma membrane,which mainly depended on hydrolysis of phospholipase D.By contrast,heat stress induced nuclear translocation of PA indicated by GFP-N160RbohD through a process that required diacylglycerol kinase activity,as well as secretory and endocytic trafficking.Strikingly,we showed that gravity triggers asymmetric PA distribution at the root apex,a response that is suppressed by PLDz2 knockout.The broad utility of the PA sensor will expand our mechanistic understanding of numerous lipid-associated physiological and cell biological processes and facilitate screening for protein candidates that the synthesis,transport,and metabolism of PA.

关 键 词：PHOSPHOLIPID phosphatidic acid RbohD heat stress SALINITY abscisic acid 

分 类 号：TP212[自动化与计算机技术—检测技术与自动化装置]