Phosphoinositide‐specific phospholipase C9 is involved in the thermotolerance of Arabidopsis

Summary Intracellular calcium (Ca2+) increases rapidly after heat shock (HS) in the Ca2+/calmodulin (Ca2+/CaM) HS signal transduction pathway: a hypothesis proposed based on our previous findings. However, evidence for the increase in Ca2+ after HS was obtained only through physiological and pharmac...

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Veröffentlicht in:The Plant journal : for cell and molecular biology 2012-02, Vol.69 (4), p.689-700
Hauptverfasser: Zheng, Shu‐Zhi, Liu, Yu‐Liang, Li, Bing, Shang, Zhong‐ lin, Zhou, Ren‐Gang, Sun, Da‐Ye
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container_title The Plant journal : for cell and molecular biology
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creator Zheng, Shu‐Zhi
Liu, Yu‐Liang
Li, Bing
Shang, Zhong‐ lin
Zhou, Ren‐Gang
Sun, Da‐Ye
description Summary Intracellular calcium (Ca2+) increases rapidly after heat shock (HS) in the Ca2+/calmodulin (Ca2+/CaM) HS signal transduction pathway: a hypothesis proposed based on our previous findings. However, evidence for the increase in Ca2+ after HS was obtained only through physiological and pharmacological experiments; thus, direct molecular genetic evidence is needed. The role of phosphoinositide‐specific phospholipase C (PI‐PLC) is poorly understood in the plant response to HS. In this work, atplc9 mutant plants displayed a serious thermosensitive phenotype compared with wild‐type (WT) plants after HS. Complementation of atplc9 with AtPLC9 rescued both the basal and acquired thermotolerance phenotype of the WT plants. In addition, thermotolerance was even improved in overexpressed lines. The GUS staining of AtPLC9 promoter:GUS transgenic seedlings showed that AtPLC9 expression was ubiquitous. The fluorescence distribution of the fusion protein AtPLC9 promoter:AtPLC9:GFP revealed that the subcellular localization of AtPLC9 was restricted to the plasma membrane. The results of a PLC activity assay showed a reduction in the accumulation of inositol‐1,4,5‐trisphosphate (IP3) in atplc9 during HS and improved IP3 generation in the overexpressed lines. Furthermore, the heat‐induced increase in intracellular Ca2+ was decreased in atplc9. Accumulation of the small HS proteins HSP18.2 and HSP25.3 was downregulated in atplc9 and upregulated in the overexpressed lines after HS. Together, these results provide molecular genetic evidence showing that AtPLC9 plays a role in thermotolerance in Arabidopsis.
doi_str_mv 10.1111/j.1365-313X.2011.04823.x
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However, evidence for the increase in Ca2+ after HS was obtained only through physiological and pharmacological experiments; thus, direct molecular genetic evidence is needed. The role of phosphoinositide‐specific phospholipase C (PI‐PLC) is poorly understood in the plant response to HS. In this work, atplc9 mutant plants displayed a serious thermosensitive phenotype compared with wild‐type (WT) plants after HS. Complementation of atplc9 with AtPLC9 rescued both the basal and acquired thermotolerance phenotype of the WT plants. In addition, thermotolerance was even improved in overexpressed lines. The GUS staining of AtPLC9 promoter:GUS transgenic seedlings showed that AtPLC9 expression was ubiquitous. The fluorescence distribution of the fusion protein AtPLC9 promoter:AtPLC9:GFP revealed that the subcellular localization of AtPLC9 was restricted to the plasma membrane. The results of a PLC activity assay showed a reduction in the accumulation of inositol‐1,4,5‐trisphosphate (IP3) in atplc9 during HS and improved IP3 generation in the overexpressed lines. Furthermore, the heat‐induced increase in intracellular Ca2+ was decreased in atplc9. Accumulation of the small HS proteins HSP18.2 and HSP25.3 was downregulated in atplc9 and upregulated in the overexpressed lines after HS. 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Psychology ; Gene expression ; Gene Expression - genetics ; Gene Expression Regulation, Plant - genetics ; Genotype &amp; phenotype ; heat shock ; Heat-Shock Proteins - metabolism ; Heat-Shock Response - physiology ; Hot Temperature ; Inositol 1,4,5-Trisphosphate - metabolism ; Molecular and cellular biology ; Molecular biology ; Mutagenesis, Insertional ; Mutation ; Phenotype ; Phosphatidylinositols - metabolism ; Phospholipases - genetics ; Phospholipases - metabolism ; Plant biology ; Plant physiology and development ; Plants, Genetically Modified - genetics ; Plants, Genetically Modified - metabolism ; Promoter Regions, Genetic - genetics ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - metabolism ; Seedlings - genetics ; Seedlings - metabolism ; Seedlings - physiology ; Signal Transduction ; Survival Analysis ; thermotolerance</subject><ispartof>The Plant journal : for cell and molecular biology, 2012-02, Vol.69 (4), p.689-700</ispartof><rights>2011 The Authors. 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However, evidence for the increase in Ca2+ after HS was obtained only through physiological and pharmacological experiments; thus, direct molecular genetic evidence is needed. The role of phosphoinositide‐specific phospholipase C (PI‐PLC) is poorly understood in the plant response to HS. In this work, atplc9 mutant plants displayed a serious thermosensitive phenotype compared with wild‐type (WT) plants after HS. Complementation of atplc9 with AtPLC9 rescued both the basal and acquired thermotolerance phenotype of the WT plants. In addition, thermotolerance was even improved in overexpressed lines. The GUS staining of AtPLC9 promoter:GUS transgenic seedlings showed that AtPLC9 expression was ubiquitous. The fluorescence distribution of the fusion protein AtPLC9 promoter:AtPLC9:GFP revealed that the subcellular localization of AtPLC9 was restricted to the plasma membrane. 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However, evidence for the increase in Ca2+ after HS was obtained only through physiological and pharmacological experiments; thus, direct molecular genetic evidence is needed. The role of phosphoinositide‐specific phospholipase C (PI‐PLC) is poorly understood in the plant response to HS. In this work, atplc9 mutant plants displayed a serious thermosensitive phenotype compared with wild‐type (WT) plants after HS. Complementation of atplc9 with AtPLC9 rescued both the basal and acquired thermotolerance phenotype of the WT plants. In addition, thermotolerance was even improved in overexpressed lines. The GUS staining of AtPLC9 promoter:GUS transgenic seedlings showed that AtPLC9 expression was ubiquitous. The fluorescence distribution of the fusion protein AtPLC9 promoter:AtPLC9:GFP revealed that the subcellular localization of AtPLC9 was restricted to the plasma membrane. The results of a PLC activity assay showed a reduction in the accumulation of inositol‐1,4,5‐trisphosphate (IP3) in atplc9 during HS and improved IP3 generation in the overexpressed lines. Furthermore, the heat‐induced increase in intracellular Ca2+ was decreased in atplc9. Accumulation of the small HS proteins HSP18.2 and HSP25.3 was downregulated in atplc9 and upregulated in the overexpressed lines after HS. Together, these results provide molecular genetic evidence showing that AtPLC9 plays a role in thermotolerance in Arabidopsis.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>22007900</pmid><doi>10.1111/j.1365-313X.2011.04823.x</doi><tpages>12</tpages></addata></record>
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subjects Acclimatization - physiology
Arabidopsis
Arabidopsis - enzymology
Arabidopsis - genetics
Arabidopsis - physiology
Arabidopsis Proteins - genetics
Arabidopsis Proteins - metabolism
AtPLC9
Biological and medical sciences
Calcium - analysis
Calcium - metabolism
Cell Membrane - enzymology
Cell physiology
Enzymes
Fundamental and applied biological sciences. Psychology
Gene expression
Gene Expression - genetics
Gene Expression Regulation, Plant - genetics
Genotype & phenotype
heat shock
Heat-Shock Proteins - metabolism
Heat-Shock Response - physiology
Hot Temperature
Inositol 1,4,5-Trisphosphate - metabolism
Molecular and cellular biology
Molecular biology
Mutagenesis, Insertional
Mutation
Phenotype
Phosphatidylinositols - metabolism
Phospholipases - genetics
Phospholipases - metabolism
Plant biology
Plant physiology and development
Plants, Genetically Modified - genetics
Plants, Genetically Modified - metabolism
Promoter Regions, Genetic - genetics
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Seedlings - genetics
Seedlings - metabolism
Seedlings - physiology
Signal Transduction
Survival Analysis
thermotolerance
title Phosphoinositide‐specific phospholipase C9 is involved in the thermotolerance of Arabidopsis
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