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dc.contributorYuki Nakamuraen_US
dc.contributor.authorHai Anh Ngoen_US
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dc.description.abstractPhospholipases play crucial roles in plant membrane lipid signaling and homeostasis. Non-specific phospholipase C (NPC) is a unique class of phospholipase found only in plants and certain bacteria. NPCs catalyze the hydrolysis of primary membrane phospholipid classes including phosphatidylcholine (PC) and phosphatidylethanolamine (PE) to release diacylglycerol (DAG) and the corresponding phosphoaminoalcohol. Six isoforms of NPCs were found in Arabidopsis thaliana, designated NPC1-NPC6. Although the physiological roles for most NPC isoforms in stress response have been described, function of NPC2 and NPC6 is unkown. In addition, for characterized NPC isoforms, molecular mechanism by which these NPCs mediate the stress response remain elusive. In this thesis, I report on our investigation about NPC2 and NPC6 in gametophyte development (Chapter 2) and NPC3 in ER stress response (Chapter 3). In Chapter 2, we showed that NPC2 and NPC6 are required for male and female gametophyte development in Arabidopsis. We found that npc2-1/- npc6-2/- was not retrieved due to the lethal effect. Both NPC2 and NPC6 proteins are localized to the plastids. Promoter-GUS assays in transgenic Arabidopsis revealed that NPC2 and NPC6 are preferentially expressed in floral organs rather than in leaves. In vitro enzyme assays showed that NPC2 and NPC6 hydrolyze phosphatidylcholine and phosphatidylethanolamine, but not phosphatidate. Lipidomic analysis of npc2-1/- npc6-2/+ and npc2-1/+ npc6-2/- showed increased contents of PC, PE, and PG in floral buds but not in mature flowers. Thus, our results indicate that NPC2 and NPC6 are functional NPCs involved in gametophyte development and glycerolipid metabolism in floral buds of A. thaliana. On the other hand, lipid metabolism is altered during endoplasmic reticulum (ER) stress in animal cells. In plants, however, was still little known. In Chapter 3, we showed that a T-DNA insertional mutant of NPC3, designed npc3-3, was hyposensitivity to Tunicamycin (TM)-induced-ER stress treatment compare to the wild-type plants. Under normal growth condition, we found that expression levels of ER stress response genes were higher in the npc3-3 mutant than that of the wild type. NPC3 has phospholipase C activity for PC and PE. Confocal microscopy observation of the subcellular localization of NPC3 showed that NPC3 was localized to the ER under normal condition but was found in nucleus after TM treatment. Our data demonstrated the requirement and nuclear translocation of NPC3 in response to ER stress. Taken together, in Chapter 4, we summarized our major findings on the NPCs, discussed on the differential roles of NPCs, provided testable hypothesis, and experimental plans. In summary, our findings described in this thesis conceptually advance our understanding on the function of NPCs in Arabidopsisen_US
dc.description.tableofcontentsAcknowledgements i Abstract iii List of Tables vi List of Figures vii List of Abbreviations ix CHAPTER 1 1 General Introduction 1 CHAPTER 2 18 A Pair of Non-specific Phospholipases C, NPC2 and NPC6, is Involved in Gametophyte Development and Glycerolipid Metabolism in Arabidopsis 18 Abstract 19 Introduction 20 Materials and Methods 23 Discussion 36 CHAPTER 3 65 Endoplasmic Reticulum-Nuclear Translocation of Non-specific Phospholipase C3 Involved in Endoplasmic Reticulum Stress Response in Arabidopsis thaliana 65 Abstract 66 Introduction 67 Materials and Methods 69 Results 74 Discussion 78 CHAPTER 4 90 Conclusion and Future Perspectives References 97 Resume 105zh_TW
dc.subjectArabidopsis thalianaen_US
dc.subjectnon-specific phospholipase Cen_US
dc.subjectGametophyte developmenten_US
dc.subjectER stressen_US
dc.title阿拉伯芥非特異性磷脂酶 (NPC)在生長發育過程之功能研究zh_TW
dc.titleA study on the function of non-specific phospholipase Cs (NPCs) in growth and development in Arabidopsis thalianaen_US
dc.typethesis and dissertationen_US
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