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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 靳宗洛 | |
dc.contributor.author | Dan-Li Luo | en |
dc.contributor.author | 羅丹利 | zh_TW |
dc.date.accessioned | 2021-06-15T06:52:21Z | - |
dc.date.available | 2016-02-20 | |
dc.date.copyright | 2011-02-20 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-02-14 | |
dc.identifier.citation | Alexandrov V (1994) Functional aspects of cell response to heat shock. International Review of Cytology 148: 171–227
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/48317 | - |
dc.description.abstract | 在乾旱或缺水等等的逆境環境下,植物會誘導細胞壁蛋白及具分解細胞壁功能的酵素,在細胞壁中扮演重要的角色。細胞壁果膠甲基酯酶(Pectin Methylesterase, PME)為一種普遍存在於細菌、真菌及植物的酵素,對植物細胞壁果膠進行去甲基酯化作用(demethylesterification),會改變果膠構造影響植物細胞壁之堅固程度。目前研究顯示,單子葉植物水稻及雙子葉植物黃豆,在熱休克(HS)誘導下能藉由調節PME之活性,參與細胞壁之重建並增強細胞間之黏結作用,恢復植物抵抗高溫之能力。分析阿拉伯芥基因組,顯示至少具有66個編譯PME的基因,本研究以正向遺傳(forward genetics)的方式,針對阿拉伯芥53個T-DNA插入造成PME基因缺失之突變株,進行熱逆境處理之篩選。結果顯示,PME34 (At3g49220)基因的缺失,明顯地減少其誘導耐熱性(acquired thermotolerance),存活率下降50%。以阿拉伯芥原生質體及洋蔥表皮細胞短暫表現PME34,證實其位於原生質膜上;另外,切除N端跨膜區域後PME34呈現點狀分散於細胞質。HS處理後,pme34突變株之PME及聚半乳醣醛酸酵素(PG)酵素活性均較野生型高,推測會加速細胞壁結構之分解而降低突變株之耐熱性。在pme34中研究與HS相關基因的表現,顯示PME34可能會受到AtHas32和AtHsfA3的調控。本研究以遺傳的層次,證實這個特殊的PME34對於植物誘導耐熱性之獲得扮演重要的角色,因此植物可藉由PME活性之調節來反應外在環境之變化。 | zh_TW |
dc.description.abstract | Pectin modification is catalyzed by a large enzyme family of pectin methylesterase (PME) residing in the cell wall. PME might modify the structure of pectin in order to adjust the characteristics of cell wall by pectin demethylesterification. In our previous study, we found that the activity of PME is induced by heat shock (HS), we further established the role of PME by retained plasma membrane integrity and coordinated with heat shock response (HSP) to confer acquired thermotolerance in planta. At least potential 66 members of PME genes are annotated in the Arabidopsis genome by a phylogenetic analysis. We tested 53 AtPME T-DNA insertion lines by thermotolerance assays, two mutants allele of AtPME34 (At3g49220) were characterized and revealed a consistent reduction in the acquired thermotolerance. The plasma membrane-localized of AtPME34 was transiently expressed in Arabidopsis protoplasts and onion cells. However, AtPME34 lacking the transmembrane domain showed the punctuated shape structures in the cytoplasm. We confirm again that the action of PME and PG during HSR might have a pronounced effect on the development of acquired thermotolerance. Furthermore, we demonstrated that the transcript of AtPME34 is probably regulated by HS–related genes such as AtHsa32 and AtHSFA3 involved in the HS response (HSR). In this study, we provide a genetic evidence, address that the unique AtPME34 plays a role during HSR to confer the acquisition of thermotolerance. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T06:52:21Z (GMT). No. of bitstreams: 1 ntu-100-R97b42018-1.pdf: 6522868 bytes, checksum: 27af4134c94eb21474b01a42de2e167b (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | ABSTRACT 4
In English …...4 In Chinese ……………………5 Abbreviations 6 Chapter 1: Introduction 7 1.1 Heat Shock Response 7 1.2 Pectin Structure 8 1.3 Pectin Methylesterase 10 1.4 Pectin Methylesterase and Inhibitor 12 1.5 The Physiology Roles of Pectin Methylesterase 13 1.6 Purpose of Research 14 Chapter 2: Materials and Methods 16 2.1 Plant Materials and Growth Condition 16 2.2 Identification of PME Insertion Mutant Lines 16 2.3 Thermotolerance Test 16 2.4 Plasmid DNA Isolation from E.coli 17 2.5 Plant Transformation 18 2.6 RNA Isolation, cDNA Synthesis and (RT)-PCR in Rice Root 18 2.7 Abiotic Stress and Hormone Treatments 19 2.8 Expression of AtPME34 in E. coli 20 2.9 Determination of PME Enzymatic Activity 20 2.10 Polygalacturonase Activity Assay 21 2.11 Subcellular Localization of AtPME34 Fusion Protein in Onion Epidermal Cell 22 2.12 Arabidopsis Protoplast Isolation and Transfection 22 Chapter 3: Results 24 3.1 Screening of AtPME T-DNA Insertion Mutants 24 3.2 Characterization of AtPME34 T-DNA Insertion Mutants 25 3.3 Expression of AtPME34 Was Reduced under Different Stress Conditions 25 3.4 Amino Acid Sequence Alignment of Arabidopsis PME Genes and Differential Expression at Development Stage of Plant by Microarray Database 26 3.5 Crude Extract from AtPME34 Expressing Bacterial Cultures Showed form Inclusion Body Formation 26 3.6 PME Activity of pme34 Mutant in Response to Heat Stress 27 3.7 PG Activity of pme34 Mutant and Wild-type under Heat Stress 27 3.8 Subcellular Localization of AtPME34 Protein in Arabidopsis Protoplast and Onion Epidermal Cells 27 3.9 Overexpressing PME34 in Arabidopsis Show the Increased PME Activity 30 3.10 Transcript of Heat Shock Transcriptional Factor A3 Was Increased in pme34 30 Chapter 4: Discussion 57 4.1 Specific Arabidopsis PME Genes Related to Heat Shock Response by Forward Genetic Analysis 57 4.2 PME Genes Reveal Dissimilarity by Microarray Database and Aligment 58 4.3 The activity of PME and PG during HSR Might Have a Pronounced Effect on the Development of Heat Tolerance 59 4.4 Transmembrane Domain of AtPME34 Is Required for Protein Targeting 61 4.5 The Defective of Heat-Related Genes Was Increased AtPME34 Transcript Levels 63 Conclusions and Prospects .. 64 Tables and Figures 32 Table 1. The lists of 53 PME knock-out mutants 32 Figure 1. Identification of Arabidopsis PME-KO mutants by thermotolerance test 34 Figure 2. Identification of Arabidopsis PME-KO mutants by thermotolerance test 35 Figure 3. Identification of Arabidopsis PME-KO mutants by thermotolerance test 36 Figure 4. Identification of Arabidopsis PME-KO mutants by thermotolerance test 37 Figure 5. Identification of Arabidopsis PME-KO mutants by thermotolerance test 38 Figure 6. Identification of Arabidopsis PME-KO mutants by thermotolerance test 39 Figure 7. Identification of Arabidopsis PME-KO mutants by thermotolerance test 40 Figure 8. Identification of Arabidopsis PME-KO mutants by thermotolerance test 41 Figure 9. The defective pme23, pme28, and pme34 mutant lines are loss acquired thermotolerance 42 Figure 10. The identification of AtPME34 T-DNA insertion line 44 Figure 11. The transcript levels of AtPME7, 28 and 34 genes were affected by plant hormone and abiotic stresses treatments 45 Figure 12. The aligment of amino acid sequences of AtPME7, AtPME34, and AtPME28 47 Figure 13. Overexpression of AtPME34 in Escherichia coli 48 Figure 14. The activity of PME and PG activity on pme28 and pme34 mutant lines under acquired thermotolerance condition 49 Figure 15. Subcellur localization of AtPME34 in Arabidopsis protoplasts 51 Figure 16. Transient expression of AtPME34 in onion epidermal cell by particle bombardment 53 Figure 17. The overexpresion lines of AtPME34 in Arabidopsis 54 Figure 18. The correlation between the expression of AtPME34 and Arabidopsis heat shock factors genes in pme34 and heat-related mutants 55 References 66 Appendixes 75 Appendix 1A. Plant cell wall and structure of pectin 75 Appendix 1B. Plant cell wall and structure of pectin 76 Appendix 1C. Plant cell wall and structure of pectin 77 Appendix 2A. The three type pectin-degrading enzymes are pectin methylesterase, polygalacturonase, and pectate lyase 78 Appendix 2B. Arabidopsis PMEs can be subdivided into two types 79 Appendix 3. A Hypothetical model of pectin modification in the pollen tube (A) and Group II PME processing (B), respectively 80 Appendix 4. The putative roles of PME genes analyze by microarray 81 Appendix 5. Construst map of pRTL2 for AtPME34 for the cloning of AtPME34 tests the subcellular localization of AtPME34 in plant cells 82 Appendix 6. The map of p2FGW7 for the cloning of AtPME34 tests the subcellular localization of AtPME34 in plant cells 83 Appendix 7. The map of pCambia3300 for the cloning of AtPME34 that was overexpressed in Arabidopsis. 84 Appendix 8. Construst map of pGEX-6P-1 for AtPME34 overexpression in E. coli. ……… 85 Appendix 9. The transcript level of AtPME7 (At1g02810) gene in the different aspects of plant development 86 Appendix 10. The transcript level of AtPME28 (At5g27870) gene in the aspects of plant development 87 Appendix 11. AtPME34 (At3g49220) gene is conserved expressed at different aspects of plant development 88 Appendix 12. The alignment of amino acid sequences of AtPME34 and AtPME34-iso2 89 Appendix 13. The list of primers 90 | |
dc.language.iso | en | |
dc.title | 阿拉伯芥果膠甲基酯酶34基因在熱逆境下之功能性研究 | zh_TW |
dc.title | Functional Study of a Pectin Methylesterases Gene, AtPME34, in Response to Heat Stress in Arabidopsis | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 林彩雲,林秋榮,常怡雍,張孟基 | |
dc.subject.keyword | 耐熱性,果膠甲基酯化酶,熱休克, | zh_TW |
dc.subject.keyword | Thermotolerance,Pectin Methylesterase,Heat Shock, | en |
dc.relation.page | 90 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-02-14 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 植物科學研究所 | zh_TW |
顯示於系所單位: | 植物科學研究所 |
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