Studies on double fertilisation and early embryogenesis of Arabidopsis thaliana Heynh. and Zea mays L.:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2012
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| Schlagworte: | |
| Online-Zugang: | kostenfrei https://nbn-resolving.org/urn:nbn:de:bvb:355-epub-257918 Inhaltsverzeichnis |
| Beschreibung: | IV, 177 S. Ill., graph. Darst. |
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Datensatz im Suchindex
| _version_ | 1804149473032536065 |
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| adam_text | IMAGE 1
CONTENTS
CHAPTER 1 - GENERAL INTRODUCTION: SEXUAL REPRODUCTION IN PLANTS I
I. I DEVELOPMENT AND FUNCTION O F THE FEMALE GAMETOPHYTE IN ANGIOSPERMS
I
1.2 DEVELOPMENT AND FUNCTION O F THE MALE GAMETOPHYTE IN ANGIOSPERMS 5
1.3 GENETIC AND HORMONAL CONTROL O F EMBRYO DEVELOPMENT IN MAIZE 6
1.4 A I M S O F THE W O R K 9
CHAPTER 2 - SPOROPHYTIC CONTROL O F POLLEN TUBE GROWTH AND GUIDANCE IN
MAIZE 11
2.1 INTRODUCTION I I
2.2 EXPERIMENTAL PROCEDURES 14
2.2.1 PLANT MATERIAL, POLLINATION AND SAMPLE PREPARATION 14
2.2.1.1 PLANT GROWTH 14
2.2.1.2 POLLINATION AND SAMPLE PREPARATION I S
2.2.2 ANILINE BLUE STAINING (MODIFIED AFTER MARTIN 1959) AND MICROSCOPY
15
2.2.3. HISTOLOGICAL STUDIES O F FIXED AND CLEARED OVULES 15
2.3 RESULTS 16
2.3.1 POLLEN GERMINATION EFFICIENCY 16
2.3.2 POLLEN TUBE GROWTH RANGE IN MAIZE AND TRIPSACWN DACTYLOIDES SILKS
18
2.3.4 PT GUIDANCE TOWARDS THE OVULAR CAVITY AND MICROPYLC 23
2.3 DISCUSSION 27
2.5 SUMMARY 32
CHAPTER 3 - CALCIUM SIGNALLING DURING DOUBLE FERTILISATION ILL
ARABIDOPSIS LHALIANA OVUIES33
3.1 INTRODUCTION 33
3.1.1 CALCIUM SIGNALS 33
3.1.2 PROTEINS INVOLVED IN CALCIUM SIGNALLING 34
3.1.3 CALCIUM SIGNALLING IN PLANT LIFE 37
3.1.4 A I M O F THIS W O R K 39
3.2 EXPERIMENTAL PROCEDURES 4 0
3.2.1 ARABIDOPSIS LHALIANA MATERIAL AND GROWTH CONDITIONS 4 0
3.2.1.1 PLANT GROWTH 4 0
3.2.1.2 STABLE TRANSFORMATION O F A. THALIANA 4 1
3.2.1.3 ASSAY O F TRANSGENE EXPRESSION 4 1
3.2.1.4 GENOMIC DNA ISOLATION 42
3.2.2 TOBACCO GROWTH CONDITIONS AND TRANSIENT TRANSGENE EXPRESSION 42
3.2.3 BACTERIAL MATERIAL, TRANSFORMATION AND GROWTH CONDITIONS 43
3.2.4 PCR AND MOLECULAR CLONING 4 4
3.2.4.1 PCR AND DNA-MODIFYING REACTIONS 44
I
HTTP://D-NB.INFO/1027030491
IMAGE 2
3.2.4.2 CLONING STRATEGIES 45
3.2.5 FLUORESCENCE MICROSCOPY 4 6
3.2.6 CONFOCAL MICROSCOPY, FRET-ASSAY AND DATA PROCESSING 47
3.3 RESULTS 48
3.3.1 EXPRESSION O F THE PLASMA MEMBRANE MARKER A C P M G IN TOBACCO AND
ARABIDOPSIS THALIANA 48
3.3.2 EXPRESSION O F THE RECOMBINANT CALCIUM SENSOR C E R T N - L I 5 IN
VEGETATIVE AND GAMETOPHYTIC CELL TYPES O F ARABIDOPSIS THALIANA 50
3.3.3 I N VITRO GROWTH O F CERTN-POLLEN TUBES AND FRET-MEASUREMENT 53
3.4 DISCUSSION 55
3.5 SUMMARY 57
CHAPTER 4 - EGG CELL SIGNALING B Y Z M E A L L CONTROLS ANTIPODAL CELL
FATE 58
4.1 INTRODUCTION 58
4.2. EXPERIMENTAL PROCEDURES 6 0
4.2.1 PLANT MATERIAL 6 0
4.2.2 HISTOLOGICAL STUDIES, IMMUNOSTAINING AND EGFP IMAGING 6 0
4.2.3 ISOLATION O F MALE AND FEMALE GAMETOPHYTIC CELLS FROM MAIZE 61
4.2.4 PLASMOLYSIS AND PCIB TREATMENT 61
4.2.5 GENERATION O F CONSTRUCTS AND STABLE MAIZE TRANSFORMATION 62
4.2.6 SEMI-QUANTITATIVE SINGLE CELL RT-PCR (SC RT-PCR) 63
4.2.7 D N A AND R N A EXTRACTION, SOUTHERN BLOT ANALYSIS AND RT-PCR 63
4.2.8 TRANSIENT TRANSFORMATION O F MAIZE B M S CELLS 64
4.2.9 QUANTITATIVE REAL-TIME PGR (QRT-PCR) 65
4.2.10 LMMUNOCYTOCHEMISTRY 66
4.3 RESULTS 67
4.3.1 ACTIVATION O F Z M E A L L AT THE MICROPYLAR POLE O F THE SYNCYLIC
EMBRYO SAC OCCURS INDEPENDENT FROM AUXIN 67
4.3.2 Z M E A L L ENCODES AN EGG CELL SECRETED E A L - B O X PEPTIDE 69
4.3.3 Z M E A L L ACTIVITY IS REQUIRED TO PREVENT ANTIPODAL CELLS FROM
ADOPTING CENTRAL CELL FATE 75
4.3.4 Z M E A L L ACTIVITY IS REQUIRED F O R I G I EXPRESSION 78
4.4 DISCUSSION 78
4.5 SUMMARY 80
CHAPTER 5 - CHARACTERISATION O F THE SIGNALLING PEPTIDE Z M E A L L IN
EMBRYO DEVELOPMENT AND MAIZE CELL SUSPENSION CULTURE 81
5.1 INTRODUCTION 81
5.2 EXPERIMENTAL PROCEDURES 84
5.2.1 ZEA MAYS L. MATERIAL PREPARATION 84
5.2.1.1 PREPARATION O F GENOMIC DNA F R O M MAIZE 84
I I
IMAGE 3
5.2.1.2 PREPARATION O F TOTAL RNA F R O M MAIZE MATERIAL AND DNASE
TREATMENT 85
5.2.1.3 PREPARATION O F PROTEINS F R O M MAIZE 86
5.2.2 NICOTIANA BENTHAMIANA TRANSIENT TRANSGENE EXPRESSION, MATERIAL
PREPARATION AND
PROTEIN PURIFICATION 86
5.2.3 BACTERIA CULTURE AND PROTEIN PURIFICATION FROM E. C O L I 87
5.2.4 YEAST CULTURE, MATERIAL PREPARATION AND YEAST-2-HYBRID EXPERIMENTS
89
5.2.4.1 YEAST CULTURE AND STRAINS 89
5.2.4.2 YEAST TRANSFORMATION 89
5.2.4.3 PLASMID PREPARATION F R O M YEAST 90
5.2.4.4 PREPARATION O F GENOMIC DNA FROM YEAST 9 1
5.2.4.5 YEAST-2-HYBRID EXPERIMENTS 9 1
5.4.2.6 COMPLEMENTATION O F SECL8 ,$ G R O W T H PHENOTYPE BY
Y2H-CANDIDATE #33 92
5.2.5 B M S CELL EULTURE AND TRANSIENT EXPRESSION O F FLUORESCENCE
PROTEIN FUSIONS 92
5.2.6 ACRIFLAVINE STAINING AND MICROSCOPY 93
5.2.7 MOLECULAR CLONING 94
5.2.7.1 TRANSIENT EXPRESSION O F AFFINITY TAGGED EA1 AND EAL1 PROTEINS
IN TOBACCO 94
5.2.7.2 HETEROLOGOUS EXPRESSION IN E. COLI 95
5.2.7.3 YEAST EXPRESSION VECTORS 96
5.2.8 PCR-BASED TRANSCRIPT LEVEL ANALYSIS 97
5.2.9 MICROARRAY ANALYSIS 98
5.2.10 IN VITRO TRANSCRIPTION/TRANSLATION AND CO-IMMUNE PRECIPITATION
(COIP) 98
5.2.11 WESTERN BLOT ANALYSIS 99
5.3 RESULTS 100
5.3.1 HETEROLOGOUS EXPRESSION O F RECOMBINANT Z M E A L AND Z M E A L L
100
5.3.1.1 EXPRESSION O F ZMEAL AND ZMEALL IN NICOTIANA BENTHAMIANA 100
5.3.1.2 EXPRESSION O F ZMEALL IN E. COLI 106
5.3.2 IDENTIFICATION O F INTERACTION PARTNERS O F I R E A L L 108
5.3.2.1 Y2H-EXPERIMENTS ON FRAGMENTS AND FULL LENGTH CLONES O F
INTERACTOR CANDIDATES.. 110
5.3.2.2 CO-IMMUNE PRECIPITATION (COIP) O F TREALL AND FULL-LENGTH
INTERACTOR CANDIDATES. 112
5.3.2.3 CHARACTERISATION O F INTERACTOR CANDIDATE #33 113
5.3.3 PATTERN FORMATION AND THE ROLE O F Z M E A L L IN EARLY EMBRYO
DEVELOPMENT 120
5.3.3.1 EXPRESSION O F MARKER CONSTRUCTS IN FERTILIZATION PRODUCTS O F
Z. MAYS 120
5.3.4.2 ANALYSIS O F EO/I-RNAI LINES DURING EARLY EMBRYO DEVELOPMENT 123
5.3.5 IDENTIFICATION O F DOWNSTREAM TARGETS O F E A L L IN B M S CELLS
126
5.3.5.1 PHENOTYPIC ANALYSIS O F BMS CELLS INCUBATED W I T H TREALL 126
5.3.5.2 MICROARRAY ANALYSIS O F BMS CELLS INCUBATED W I T H TREALL 127
III
IMAGE 4
5.4 DISCUSSION 136
5.4.1 INTERACTION PARTNER O F T R E A L I 136
5.4.3 PATTERN FORMATION AND REGULATION O L Z M E A L L B Y AUXIN 137
5.4.5 DOWNSTREAM TARGETS O F Z M E A L L SIGNALLING 140
5.4.6 OUTLOOK 141
5.5 SUMMARY 142
CHAPTER 6 - COMPREHENSIVE DISCUSSION AND OUTLOOK 144
REFERENCES 148
SUPPLEMENTARY INFORMATION 163
ANTIBIOTICS, ANTIBODIES AND PRIMER 163
VECTOR MAPS 166
ABBREVIATIONS 175
ACKNOWLEDGEMENTS 177
IV
|
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| spelling | Lausser, Andreas Verfasser (DE-588)1027030394 aut Studies on double fertilisation and early embryogenesis of Arabidopsis thaliana Heynh. and Zea mays L. vorgelegt von Andreas Lausser 2012 IV, 177 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Regensburg, Univ., Diss., 2012 Embryonalentwicklung (DE-588)4070792-1 gnd rswk-swf Mais (DE-588)4037135-9 gnd rswk-swf Ackerschmalwand (DE-588)4141299-0 gnd rswk-swf Befruchtung (DE-588)4005234-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Ackerschmalwand (DE-588)4141299-0 s Mais (DE-588)4037135-9 s Befruchtung (DE-588)4005234-5 s Embryonalentwicklung (DE-588)4070792-1 s b DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:355-epub-257918 http://epub.uni-regensburg.de/25791/ Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:355-epub-257918 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025269199&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Lausser, Andreas Studies on double fertilisation and early embryogenesis of Arabidopsis thaliana Heynh. and Zea mays L. Embryonalentwicklung (DE-588)4070792-1 gnd Mais (DE-588)4037135-9 gnd Ackerschmalwand (DE-588)4141299-0 gnd Befruchtung (DE-588)4005234-5 gnd |
| subject_GND | (DE-588)4070792-1 (DE-588)4037135-9 (DE-588)4141299-0 (DE-588)4005234-5 (DE-588)4113937-9 |
| title | Studies on double fertilisation and early embryogenesis of Arabidopsis thaliana Heynh. and Zea mays L. |
| title_auth | Studies on double fertilisation and early embryogenesis of Arabidopsis thaliana Heynh. and Zea mays L. |
| title_exact_search | Studies on double fertilisation and early embryogenesis of Arabidopsis thaliana Heynh. and Zea mays L. |
| title_full | Studies on double fertilisation and early embryogenesis of Arabidopsis thaliana Heynh. and Zea mays L. vorgelegt von Andreas Lausser |
| title_fullStr | Studies on double fertilisation and early embryogenesis of Arabidopsis thaliana Heynh. and Zea mays L. vorgelegt von Andreas Lausser |
| title_full_unstemmed | Studies on double fertilisation and early embryogenesis of Arabidopsis thaliana Heynh. and Zea mays L. vorgelegt von Andreas Lausser |
| title_short | Studies on double fertilisation and early embryogenesis of Arabidopsis thaliana Heynh. and Zea mays L. |
| title_sort | studies on double fertilisation and early embryogenesis of arabidopsis thaliana heynh and zea mays l |
| topic | Embryonalentwicklung (DE-588)4070792-1 gnd Mais (DE-588)4037135-9 gnd Ackerschmalwand (DE-588)4141299-0 gnd Befruchtung (DE-588)4005234-5 gnd |
| topic_facet | Embryonalentwicklung Mais Ackerschmalwand Befruchtung Hochschulschrift |
| url | http://epub.uni-regensburg.de/25791/ https://nbn-resolving.org/urn:nbn:de:bvb:355-epub-257918 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025269199&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT lausserandreas studiesondoublefertilisationandearlyembryogenesisofarabidopsisthalianaheynhandzeamaysl |