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  2. 生命科學院
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請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60489
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor楊健志(Chien-Chih Yang)
dc.contributor.authorChia-Chun Linen
dc.contributor.author林佳君zh_TW
dc.date.accessioned2021-06-16T10:19:33Z-
dc.date.available2018-09-06
dc.date.copyright2013-09-06
dc.date.issued2013
dc.date.submitted2013-08-16
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高艾玲 (2010) 阿拉伯芥 AtMAPRs 之分子特性研究. 臺灣大學微生物與生化學研究所學位論文. 臺灣大學, pp 1-221
郭時鈺 (2012) 阿拉伯芥中 AtMAPR3 表現受 d-amino levulinic acid 及逆境誘導之研究. 臺灣大學生化科技學系學位論文. 臺灣大學, pp 1-102
dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60489-
dc.description.abstractRecombinant AtMAPR3 purified from E. coli has been shown to possess heme binding ability. AtMAPR3 expression is increased by treated with hydrogen peroxide (H2O2) or δ-amino levulinic acid (ALA), the precursor of tetrapyrroles. The ALA-induced AtMAPR3 expression is blocked by dipyridyl (DPD), an inhibitor of ferrochelatase, indicating a possible induction route via heme.
To further clarify if the “via heme” route related to the “via H2O2” route, we treated Arabidopsis with ascorbate, a H2O2 scavenger. It was surprised to find that the AtMAPR3 expression was increased by 8-fold after 6 hours treatment. This result suggested that the AtMAPR3 expression was most likely responsive to a reactive oxygen species (ROS) scavenging system. Since ALA is the precursor of several candidates of retrograde signaling molecules, whether AtMAPR3 expression was regulated by retrograde signaling was also studied. The expression of photosynthesis-associated nuclear genes (PhANGs) were monitored in wild-type, AtMAPR3 overexpression mutant (AtMAPR3-OX) and AtMAPR3 knockout mutant (AtMAPR3-KO) with or without ALA treatment. It was interesting that AtMAPR3 showed opposite expression profile to PhANGs in response to the tetrapyrrole perturbation. Further analysis concerning the role of AtMAPR3 in retrograde signaling is needed. A possible role of AtMAPR3 was proposed that it accommodated free heme and the expression of AtMAPR3 was through a ROS scavenging system triggered by a “specific ROS pool”, which was enhanced in this study by ALA treatment.
en
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Previous issue date: 2013
en
dc.description.tableofcontentsContents………………………………………………………………………………..I
Contents of figures………………………………………………………………….....IV
Abbreviation List………………………………………………………………………V
中文摘要……………………………………………………………………………..VII
Abstract……………………………………………………………………………...VIII
Chapter 1 Introduction ................................................................................................... 1
1.1 Membrane-Associated Progesterone Receptor in Arabidopsis thaliana (AtMAPR)
....................................................................................................................................... 1
1.2 ALA is the precursor of tetrapyrroles ..................................................................... 2
1.2.1 ALA is synthesized via C5-pathway in plants .................................................. 2
1.2.2 Tetrapyrroles mediate various biological processes......................................... 3
1.3 ROS is everywhere in cell ....................................................................................... 4
1.4 Ascorbate is a plentiful antioxidant in plants .......................................................... 5
1.4.1 Ascorbate is synthesized from D-glucose-6-phosphate.................................... 5
1.4.2 Ascorbate has multiple functions ..................................................................... 5
1.5 Plastid to nucleus intracellular signaling................................................................. 6
1.5.1 Historical overview of retrograde research ...................................................... 7
1.5.2 The candidates of retrograde signal molecules are generated in chloroplasts.. 7
1.6 Aims of the research................................................................................................ 9
Chapter 2 Materials and Methods ................................................................................ 10
2.1 Materials ........................................................................................................................... 10
2.1.1 Wild type.................................................................................................................... 10
2.1.2 AtMAPR3 knockout mutant ...................................................................................... 10
2.1.3 AtMAPR3 overexpression mutant ............................................................................. 10
2.1.4 AtMAPR3 promoter500::GUS ................................................................................... 11
2.2 Methods............................................................................................................................. 12
2.2.1 Plant growth condition ............................................................................................... 12
2.2.2 Chemical treatments before sampling........................................................................ 12
2.2.3 H2O2 gradient plant .................................................................................................... 13
2.2.4 Quantitative real-time PCR (qRT-PCR) .................................................................... 14
2.2.5 Histochemical analysis of β-glucuronidase (GUS) activity ....................................... 15
2.2.6 Detection of H2O2 ...................................................................................................... 16
Chapter 3 Results ......................................................................................................... 17
3.1 Ascorbate induced AtMAPR3 expression higher than ALA did ....................................... 17
3.2 Histochemistry with transgenic plant carrying pAtMAPR3::GUS.................................... 20
3.2.1 AtMAPR3::GUS histochemical analysis correlated with qRT-PCR results............... 20
3.2.2 AtMAPR3::GUS showed activity in cotyledons......................................................... 22
3.3 Is AtMAPR3 expression regulated by retrograde signaling?............................................. 23
3.4 Does AtMAPR3 mutants accumulate more H2O2? ........................................................... 24
3.5 Phenotype observation of AtMAPR3 mutants in response to oxidative stress ................. 26
Chapter 4 Discussion ................................................................................................... 27
4.1 Does AtMAPR3 remove free heme?................................................................................. 28
4.2 AtMAPR3 expression may be induced by specific “H2O2 pool” ....................................... 29
4.3 AtMAPR3 expression may be driven by heme-induced ROS scavenging system ............ 30
Chapter 5 Prospect ....................................................................................................... 32
5.1 The induction route of AtMAPR3...................................................................................... 32
5.1.1 The fate of exogenous ALA in the cell ...................................................................... 32
5.1.2 The connection between ALA, H2O2 and ascorbate on the induction of AtMAPR3
expression............................................................................................................................ 33
5.2 The physiological roles for AtMAPR3 ............................................................................. 33
5.2 The physiological roles for AtMAPR3 ............................................................................. 33
5.2.1 The cellular condition of AtMAPR3 mutants ............................................................ 33
5.2.2 Does AtMAP3 participate in the retrograde signaling? ............................................. 34
5.2.3 Possible physiological roles for AtMAPR3 in peroxisome ....................................... 34
References....................................................................................................................35
Figures………………………………………………………………………………..40
Appendices…………………………………………………………………………...54
dc.language.isoen
dc.subjectAtMAPR3zh_TW
dc.subjectδ-aminolevulinic acidzh_TW
dc.subjectAscorbatezh_TW
dc.subjectROS poolzh_TW
dc.subjectRetrogradezh_TW
dc.titleAtMAPR3 受δ-amino levulinic acid 誘導而表現之機制研究zh_TW
dc.titleStudies on the mechanism of ALA-induced AtMAPR3 expressionen
dc.typeThesis
dc.date.schoolyear101-2
dc.description.degree碩士
dc.contributor.oralexamcommittee王愛玉,李昆達,黃楓婷,廖憶純
dc.subject.keywordAtMAPR3,δ-aminolevulinic acid,Ascorbate,ROS pool,Retrograde,zh_TW
dc.relation.page55
dc.rights.note有償授權
dc.date.accepted2013-08-16
dc.contributor.author-college生命科學院zh_TW
dc.contributor.author-dept生化科技學系zh_TW
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