Regulation of respiration during low oxygen availability:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2014
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 144 Bl. Ill., graph. Darst. |
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Datensatz im Suchindex
| _version_ | 1804153209604800512 |
|---|---|
| adam_text | SUMMARY
3
ZUSAMMENFASSUNG 5
1 INTROPUCTION 12
1.1 OXYGEN GRADIENTS INSIDE PLANT TISSUES 12
1.2 OXIDATIVE PHOSPHORYLATION 14
1.3 RATE OF RESPIRATORY OXYGEN CONSUMPTION 16
1.4 METABOLIC RESPONSES OF PLANTS TO HYPOXIC CONDITIONS 17
1.4.1
METABOLIC RESPONSES OF PLANTS TO FLOODING, WATERLOGGING, AND SUBMERGENCE
18
1.4.2
METABOLIC RESPONSES TO HYPOXIA IN BULKY PLANT TISSUES AND DEVELOPING
PLANTS
21
1.5 REGULATION OF ENZYME ACTIVITY UNDER LOW OXYGEN
AVAILABILITY 23
1.5.1
ENZYMATIC ADAPTATION OF PRIMARY METABOLISM DUERING
OXYGEN LIMITATION
23
1.5.2
REGULATION OF THE MITOCHONDRIAL ELECTRON TRANSPORT CHAIN
25
1.6 HYPOTHESES OF A REGULATORY MECHANISM FOR
RESPIRATORY OXYGEN CONSUMPTION 26
1.7 CHLAMYDOMONAS REINHARDTII AS AN EXPERIMENTAL
SYSTEM FOR MITOCHONDRIAL RESPIRATION 28
1.8 ALM OFTHIS STUDY 30
2 METHODOLOGY 34
2.1 MATERIALS 34
2.1.1 CHEMICALS 34
2.1.2
EQUIPMENT
34
2.1.2.1 GENERAL LAB EQUIPMENT 34
2.1.2.2 CENTRIFUGES 35
2.1.2.3 CELL GROWTH 35
2.1.2.4 BIOREACTOR COMPONENTS 35
2.1.3 PLANT AND ALGAE MATERIAL 36
2.1.4
MEDIAAND BUFFER 36
2.1.4.1 COMPOSITION OF GROWTH MEDIA
36
2.1.5 STANDARD SOLUTIONS AND BUFFER 38
2.2 METHODS 39
2.2.1 PLANT GROWTH CONDITIONS 39
2.2.1.1 STERILE ARABIDOPSIS GROWTH
39
2.2.1.2 LIQUID CULTURES 40
2.2.2
NLTRIC OXIDE MEASUREMENTS
40
2.2.3
CONFOCAL LASER SCANNING MICROSCOPY
40
2.2.4
TRANSVERSAL ROOTSECTIONS
41
2.2.5 OXYGEN CONSUMPTION MEASUREMENT 41
2.2.6
CHLAMYDOMONAS
GROWTH CONDITIONS 43
7
HTTP://D-NB.INFO/1069586846
2.2.6.1 CULTIVATION ON PLATES 43
2.2.6.2 CULTIVATION IN FLASKS 43
2.2.6.3 CULTIVATION IN A BIOREACTOR 43
2.2.7 GLOVEBOX -A VARIABLE ATMOSPHERE WORKSTATION 45
2.2.8 MONITORING GROWTH AND CELL CYCLE (CELL COUNTING) 46
2.2.9 CELL HARVEST 46
2.2.9.1 STANDARD 47
2.2.9.2 BIOREACTOR 47
2.2.10 WHOLE CELL PROTEIN EXTRACTION 47
2.2.10.1 SDS-PAGE 48
2.2.10.2 WESTERN BLOT 48
2.2.10.3 IMMUNODETECTION OF PROTEINS 48
2.2.11 DETERMINATION OF CHLOROPHYLL 49
2.2.12 STARCHASSAY 49
2.2.13 ACETATEASSAY 49
2.2.14 ETHANOL ASSAY 50
2.2.15 METABOLITE ANALYSIS 50
2.2.15.1 EXTRACTION OF POLAR METABOLITES FOR ADAPTATION OF
CHLAMYDOMONAS TO DECREASING OXYGEN
AVAILABILITY 51
2.2.15.2 EXTRACTION OF POLAR METABOLITES FROM MEDIA 51
2.2.15.3 EXTRACTION OF POLAR METABOLITES FROM GLOVEBOX EXPERIMENT 51
2.2.15.4 METABOLITE MEASUREMENT FOR ADAPTATION OF CHLAMYDOMONAS TO
DECREASING OXYGEN
AVAILABILITY 52
2.2.15.5 METABOLITE MEASUREMENT FOR PRIMARY METABOLITES CHANGES TO
HYPOXIC CONDITIONS AND
PRIMARY METABOLITES IN EXUDATES 52
2.2.15.6 METABOLITE DATA NORMALIZATION, VISUALIZATION AND STATISTICAL
ANALYSIS 52
2.2.16 MODEUNG RESPIRATION DATA 53
2.2.16.1 MULTISCALE GAS-EXCHANGE MODEL FOR ARABIDOPSIS ROOTS 53
2.2.16.1.1 GEOMETRICAL MODEL 53
2.2.16.1.2 GAS EXCHANGE MODEL 53
2.2.16.1.3 MODEL PARAMETERS 54
2.2.16.2 MODELING CHLAMYDOMONAS RESPIRATION USING DIFFERENT
MODIFICATIONS OF MICHAELIS-MENTEN
KINETICS 55
2.2.16.2.1 MODEL 1 - CLASSICAL MICHAELIS-MENTEN MODEL 56
2.2.16.2.2 MODEL 2 - MICHAELIS-MENTEN MODEL WITH AN LINEAR EXTENSION 56
2.2.16.2.3 MODEL 3 - MICHAELIS-MENTEN MODEL WITH 0
2
REGULATED SINGLE ENZYME INACTIVATION 57
2.2.16.2.4 MODEL 4 - MICHAELIS-MENTEN MODEL WITH 0
2
REGULATED PARALLEL ENZYME INACTIVATION 58
2.2.16.2.5 MODEL 5 - KINETIC MODEL WITH 0
2
REGULATED ENZYME TRANSFORMATION 58
2.2.16.3 SENSITIVITY ANALYSIS 59
8
3 RESULTS 62
3.1 MODEUNG OXYGEN CONCENTRATION DEPENDENT RESPIRATORY ACTIVITY 62
3.1.1 DEVELOPMENT OF A MULTISCALE GAS-EXCHANGE
MODEL FOR
ARABIDOPSIS THALIANA
62
3.1.2 THE UNICELLULAR ORGANISM
CHLAMYDOMONAS REINHARDTII
SHOWS SIMILAR
RESPIRATORY PROFILE AS VASCULAR
PLANTS
64
3.1.3
VARIOUS MODEUNG APPROACHES DESCRIBE
CHLAMYDOMONAS
RESPIRATORY
ACTIVITY IN RESPONSE TO CHANGING
OXYGEN CONCENTRATION 66
3.1.3.1 MODEL 1 - CLASSICAL MICHAELIS-MENTEN KINETICS 66
3.1.3.2 MODEL
2 -
MICHAELIS-MENTEN KINETICS MODEL WITH A LINEAR EXTENSION 67
3.1.3.3 MODEL 3 - MICHAELIS-MENTEN MODEL WITH 0
2
REGULATED SINGLE ENZYME INACTIVATION 68
3.1.3.4 MODEL 4 - MICHAELIS-MENTEN MODEL WITH 0
2
REGULATED PARALLEL ENZYME INACTIVATION 69
3.1.3.5 MODEL 5 - MICHAELIS-MENTEN MODEL WITH 0
2
-REGULATED ENZYME TRANSFORMATION 70
3.1.4 BACK TO THE ROOTS -
KINETIC MODEL INCORPORATED TO THE
ARABIDOPSIS
GAS EXCHANGE MODEL 72
3.1.5 SENSITIVITY ANALYSIS 75
3.1.6 OXYGEN DISTRIBUTION IN
ARABIDOPSIS
ROOTS USING REP0
2
RTER AND AN APPLIED
BOUNDARY RESISTANCE 76
3.2 MANIPULATION OF THE OXYGEN CONSUMPTION BY THE METC OF ARABIDOPSIS
THALIANA 80
3.2.1 NLTRIC OXIDE HAS NO EFFECT ON RESPIRATION BY AFFECTING THE COX
ACTIVITY 80
3.2.2
PYRUVATE HAS AN EFFECT ON THE RESPIRATION, BUT DOES
NOT CHANGE THE CYTOLOGY OF THE ROOTS
82
3.2.3 THE RATIO COX TO AOX CAPACITY CHANGES AT LOW OXYGEN
AVAILABILITY 84
3.3 MANIPULATION OF THE OXYGEN CONSUMPTION BY THE METC IN
CHLAMYDOMONAS REINHARDTII 85
3.3.1 DIFFERENTGROWTH MEDIA AFFECT DIVERSE THE OXYGEN CONSUMPTION 85
3.3.1.1 RESPIRATORY RATE OF PHOTOAUTOTROPHICALLY GROWN CHLAMYDOMONAS 86
3.3.1.2 RESPIRATORY RATE OF MIXOTROPHICALLY GROWN CHLAMYDOMONAS 86
3.3.1.3 RESPIRATORY CAPACITY PROFILES OF TERMINAL OXIDASES IN METC 87
3.3.1.4 PYRUVATE INCREASES THE RESPIRATORY CAPACITY OF AOX 88
3.3.1.5 THE AOX PROTEIN ABUNDANCE INCREASES WHEN PYRUVATE IS ADDED 88
3.3.2 DIFFERENT GROWTH MEDIA INTERFERE IN DIFFERENT WAYS WITH THE OXYGEN
CONSUMPTION WHEN NAHC0
3
IS
ADDED 89
3.3.2.1 RESPIRATION PROFILES OF DIFFERENT GROWTH CONDITIONS INCLUDING
NAHC0
3
AS CARBON SOURCE 89
3.3.2.2 RESPIRATORY CAPACITY PROFILES OF TERMINAL OXIDASES OF METC IN
PHOTOAUTOTROPHIC CONDITIONS 90
3.3.2.3 RESPIRATORY CAPACITY PROFILES OF TERMINAL OXIDASES OF METC IN
MIXOTROPHIC CONDITIONS 91
3.3.2.4 PROTEIN ABUNDANCE OF BOTH TERMINAL OXIDASES INCREASES IN THE
PRESENCE OF PYRUVATE 92
3.4 ADAPTATION OF CHLAMYDOMONAS REINHARDTII BIOREACTOR CELL CULTURES TO
LOW OXYGEN AVAILABILITY
93
3.4.1 CHANGES IN OXYGEN CONSUMPTION AS RESPONSE TO ADAPTATION TO LOW
OXYGEN AVAILABILITY 94
3.4.2 PRIMARY METABOLITE
CHANGES
95
3.4.2.1 THE DAY / NIGHT EFFECT ON PRIMARY METABOLITES 98
3.4.2.2 THE CHANGE OF PRIMARY METABOLITES LEVELS TO LOW OXYGEN
AVAILABILITY 98
3.4.2.3 PRIMARY METABOLITES IN THE EXUDATE 100
3.4.2.4 FERMENTATION PRODUCTS 100
3.4.3 CHANGES OF GROWTH RATE AND CYTOLOGICAL PARAMETERS AS ADAPTATIONAL
RESPONSE TO LOW OXYGEN
AVAILABILITY
101
9
3.4.4 PRIMARY METABOLITES CHANGES THROUGH TIME TO HYPOXIC CONDITIONS
4 DISCUSSION
104
107
4.1 MODELLING THE OXYGEN CONSUMPTION 108
4.1.1 RESPIRATORY PROFILE
FOR
ARABIDOPSIS THAUANA
ROOTS 108
4.1.2 VARIOUS MODELS CAN DESCRIBE
CHLAMYDOMONAS
RESPIRATORY ACTIVITY IN
RESPONSE TO CHANGING OXYGEN
CONCENTRATION 109
4.1.3 BACK TO THE ROOTS -
KINETIC MODEL INCORPORATION TO THE
ARABIDOPSIS
MODEL 111
4.2 MANIPULATION OF THE OXYGEN CONSUMPTION BY THE METC 112
4.2.1
NLTRIC OXIDE
113
4.2.2 PYRUVATE 114
4.2.3 PARALALEACTIVATIONOFAOXANDCOX 116
4.2.4 MANIPULATION OF THE OXYGEN CONSUMPTION BY THE
METC IN CHLAMYDOMONAS 116
4.3 METABOLIC ADAPTATION OF CHLAMYDOMONAS REINHARDTII TO LOW OXYGEN
AVAILABILITY 118
4.3.1 HYPOXIC RESPIRATION PROFILES 119
4.3.2 METABOLIC ADAPTATION 120
4.4 SUGGESTION FOR PUTITATIVE REGULATIVE MECHANISM OF RESPIRATORY
ACTIVITY 123
5 REFERENCES 125
6 CONTENT OF ILLUSTRATIONS 139
6.1 TABLES 139
6.2
FLGURES
139
7 LIST OF ABBREVIATIONS 141
10
|
| any_adam_object | 1 |
| author | Päpke, Carola 1984- |
| author_GND | (DE-588)1067691421 |
| author_facet | Päpke, Carola 1984- |
| author_role | aut |
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| dewey-full | 572.472 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 572 - Biochemistry |
| dewey-raw | 572.472 |
| dewey-search | 572.472 |
| dewey-sort | 3572.472 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
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| spelling | Päpke, Carola 1984- Verfasser (DE-588)1067691421 aut Regulation of respiration during low oxygen availability von Carola Päpke 2014 144 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Potsdam, Univ., Diss., 2014 Atmung (DE-588)4003404-5 gnd rswk-swf Sauerstofftransport (DE-588)4179219-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Atmung (DE-588)4003404-5 s Sauerstofftransport (DE-588)4179219-1 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027915819&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Päpke, Carola 1984- Regulation of respiration during low oxygen availability Atmung (DE-588)4003404-5 gnd Sauerstofftransport (DE-588)4179219-1 gnd |
| subject_GND | (DE-588)4003404-5 (DE-588)4179219-1 (DE-588)4113937-9 |
| title | Regulation of respiration during low oxygen availability |
| title_auth | Regulation of respiration during low oxygen availability |
| title_exact_search | Regulation of respiration during low oxygen availability |
| title_full | Regulation of respiration during low oxygen availability von Carola Päpke |
| title_fullStr | Regulation of respiration during low oxygen availability von Carola Päpke |
| title_full_unstemmed | Regulation of respiration during low oxygen availability von Carola Päpke |
| title_short | Regulation of respiration during low oxygen availability |
| title_sort | regulation of respiration during low oxygen availability |
| topic | Atmung (DE-588)4003404-5 gnd Sauerstofftransport (DE-588)4179219-1 gnd |
| topic_facet | Atmung Sauerstofftransport Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027915819&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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