Characterization of genes involved in pollen development and anther opening in barley:
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|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Köln
2020
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis Inhaltsverzeichnis |
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| adam_text | TABLE OF CONTENTS
TABLE OF CONTENTS
PUBLICATION I
TABLE OF CONTENTS II
LIST OF FIGURES AND TABLES V
LIST OF ABBREVIATIONS VII
SUMMARY IX
ZUSAMMENFASSUNG XI
1. INTRODUCTION 13
1.1 STAMEN AND ANTHER DEVELOPMENT 14
1.2 POLLEN WALL FORMATION 15
1.3 TAPETUM FUNCTION AND DEVELOPMENT 16
1.4 TAPETUM DEGENERATION 18
1.5 GENETIC CONTROL OF POLLEN AND TAPETUM DEVELOPMENT 20
1.5.1 TRANSCRIPTIONAL REGULATORS 20
1.5.2 ENZYMES INVOLVED IN EXINE FORMATION 21
1.5.3 TAPETAL PCD REGULATION: TRIGGERS AND EXECUTORS 23
1.6 ANTHER OPENING 25
THESIS AIMS 27
2. RESULTS 28
CHAPTER I 28
2.1 ESTABLISHING A SCALE FOR BARLEY ANTHER DEVELOPMENT 28
2.2 PHENOTYPIC CHARACTERIZATION OF MSG32 30
2.2.1 MORPHOLOGICAL DESCRIPTION OF MSG32 30
2.2.2 HISTOLOGICAL FEATURES OF MSG32 ANTHERS IN SEMI-THIN SECTIONS 31
2.2.3 ULTRASTRUCTURAL ANALYSIS OF MSG32 ANTHERS BY TRANSMISSION ELECTRON
MICROSCOPY (TEM) 34
2.2.4 FOLLOWING DNA FRAGMENTATION AS A HALLMARK OF PCD IN BARLEY ANTHERS
49
2.3 IDENTIFICATION OF THE GENE UNDERLYING THE MALE STERILE PHENOTYPE OF
MSG32 53
2.3.1 GENETIC MAPPING OF MSG32 53
2.3.2 IDENTIFICATION OF NEW MUTANT HVALDH2B ALLELES VIA CRISPR-CAS9 54
II
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/1206379111
TABLE OF CONTENTS
2.4 FUNCTIONAL CHARACTERIZATION OF MSG32 56
2.4.1 PHYLOGENETIC ANALYSIS 56
2.4.2 EXPRESSION ANALYSIS OF HVALDH2B 58
2.4.3 COMPARATIVE TRANSCRIPTOMICS OF WILD-TYPE AND MSG32 REPRODUCTIVE
TISSUES 59
CHAPTER II 64
2.5 PHENOTYPIC CHARACTERIZATION OF MSG36.BK 64
2.5.1 MORPHOLOGICAL DESCRIPTION OF MSG36.BK 64
2.5.2 HISTOLOGICAL FEATURES OF MSG36.BK ANTHERS 65
2.6 IDENTIFICATION OF THE MSG36 GENE 66
2.7 FUNCTIONAL CHARACTERIZATION OF MSG36 69
2.7.1 EXPRESSION ANALYSIS OF MSG36.BK 69
2.7.2 STUDYING THE LEVEL OF FUNCTIONAL CONSERVATION IN PECTIN-DEGRADING
ENZYMES 70
3. DISCUSSION 72
CHAPTER I 72
3.1 MSG32 IS INVOLVED IN POST-MEIOTIC MICROSPORE DEVELOPMENT AND TAPETUM
FUNCTION 72
3.1.1 MSG32 DEFECTS APPEAR POST-MEIOTICALLY UPON MICROSPORE VACUOLATION
AND ALTER MICROSPORE
DEVELOPMENT AND TAPETUM FUNCTION 72
3.1.2 MSG32 TAPETUM DEGENERATION INITIATED ON TIME BUT WAS IMPAIRED IN
AUTOPHAGY-RELATED
PROCESSES 76
3.1.3 POTENTIAL FUNCTION OF HVALDH2B IN ANTHER DEVELOPMENT OF BARLEY 77
3.1.4 CONCLUSIONS AND FUTURE PERSPECTIVES 82
CHAPTER II 84
3.2 MSG36 IS REQUIRED FOR CELL SEPARATION PROCESSES IN ANTHER OPENING 84
3.2.1 MSG36.BK DEFECTS OCCURRED IN SEPTUM AND STOMIUM CELLS UPON
MUTATION OF HVPL 84
3.2.2 PRELIMINARY EXPRESSION PROFILE OF HVPL AND STUDIES OF ARABIDOPSIS
HOMOLOGS 85
3.2.3 CONCLUSION AND FUTURE PERSPECTIVES 86
4. MATERIAL AND METHODS 87
4.1 PLANT MATERIAL 87
4.1.1 ARABIDOPSIS 87
4.1.2 BARLEY 87
4.2 PLANT CULTIVATION 88
4.2.1 CHAMBER CULTIVATION OF ARABIDOPSIS 88
4.2.2 GREENHOUSE CULTIVATION OF BARLEY 88
4.3 PHENOTYPING/SCORING OF DEVELOPMENT 89
4.4 COMPLEMENTATION TESTS OF BARLEY 89
4.5 ARABIDOPSIS CROSSING 90
4.6 ALEXANDER S STAIN TO TEST POLLEN VIABILITY 90
4.7 SCANNING ELECTRON MICROSCOPY (SEM) 90
4.8 HISTOLOGICAL ANALYSIS 91
4.8.1 CHEMICALLY FIXED BARLEY FLOWERS 91
III
TABLE OF CONTENTS
4.8.2 BARLEY FLOWERS FIXED BY HIGH-PRESSURE FREEZING/FREEZE SUBSTITUTION
(HPF/FS) 91
4.9 ULTRASTRUCTURAL ANALYSIS OF CHEMICALLY FIXED AND HIGH-PRESSURE
FROZEN/FREEZE-SUBSTITUTED
BARLEY FLOWERS 92
4.9.1 CHEMICALLY FIXED BARLEY FLOWERS 92
4.9.2 BARLEY FLOWERS ANALYSED BY HPF/FS 93
4.10 TRANSMISSION ELECTRON MICROSCOPY (TEM) 94
4.11 POLLEN WALL MEASUREMENTS OF CHEMICALLY FIXED SAMPLES 94
4.12 DNA FRAGMENTATION DETECTION/TUNEL ASSAY 94
4.13 RNA IN SITU HYBRIDISATION OF BARLEY 96
4.14 COLLECTION OF PLANT MATERIAL FOR QRT-PCR AND TRANSCRIPTOME
SEQUENCING FOR DIFFERENTIALLY
EXPRESSED TRANSCRIPT (DET) ANALYSIS 97
4.15 TRANSCRIPTOME SEQUENCING FOR VARIANT CALLING FOR THE IDENTIFICATION
OF CANDIDATE GENES 98
4.16 RNA EXTRACTION, CDNA SYNTHESIS AND QRT-PCR 100
4.17 DNA EXTRACTION 101
4.18 MAPPING FOR CANDIDATE GENE IDENTIFICATION 102
4.19 OLIGONUCLEOTIDES 102
4.20 POLYMERASE CHAIN REACTIONS (PCR) 108
4.21 RESTRICTION ENDONUCLEASE DIGESTION AND AGAROSE GEL ELECTROPHORESIS
OF DNA 108
4.22 PLASMID CONSTRUCTION FOR GENERATION OF TRANSGENIC BARLEY PLANTS 109
4.22.1 CRISPR-CAS9 PLASMID CONSTRUCTION 109
4.22.2 PLANT TRANSFORMATION 110
4.23 SEQUENCE INFORMATION AND DISPLAY 111
4.24 IDENTIFICATION OF HOMOLOGS AND PHYLOGENETIC ANALYSIS 111
5. REFERENCES 114
6. SUPPLEMENTARY DATA 131
7. CURRICULUM VITAE 135
8. AFFLDAVIT/EIDESSTATTLICHE ERKLARUNG 137
9. ACKNOWLEDGEMENT 138
IV
|
| adam_txt |
TABLE OF CONTENTS
TABLE OF CONTENTS
PUBLICATION I
TABLE OF CONTENTS II
LIST OF FIGURES AND TABLES V
LIST OF ABBREVIATIONS VII
SUMMARY IX
ZUSAMMENFASSUNG XI
1. INTRODUCTION 13
1.1 STAMEN AND ANTHER DEVELOPMENT 14
1.2 POLLEN WALL FORMATION 15
1.3 TAPETUM FUNCTION AND DEVELOPMENT 16
1.4 TAPETUM DEGENERATION 18
1.5 GENETIC CONTROL OF POLLEN AND TAPETUM DEVELOPMENT 20
1.5.1 TRANSCRIPTIONAL REGULATORS 20
1.5.2 ENZYMES INVOLVED IN EXINE FORMATION 21
1.5.3 TAPETAL PCD REGULATION: TRIGGERS AND EXECUTORS 23
1.6 ANTHER OPENING 25
THESIS AIMS 27
2. RESULTS 28
CHAPTER I 28
2.1 ESTABLISHING A SCALE FOR BARLEY ANTHER DEVELOPMENT 28
2.2 PHENOTYPIC CHARACTERIZATION OF MSG32 30
2.2.1 MORPHOLOGICAL DESCRIPTION OF MSG32 30
2.2.2 HISTOLOGICAL FEATURES OF MSG32 ANTHERS IN SEMI-THIN SECTIONS 31
2.2.3 ULTRASTRUCTURAL ANALYSIS OF MSG32 ANTHERS BY TRANSMISSION ELECTRON
MICROSCOPY (TEM) 34
2.2.4 FOLLOWING DNA FRAGMENTATION AS A HALLMARK OF PCD IN BARLEY ANTHERS
49
2.3 IDENTIFICATION OF THE GENE UNDERLYING THE MALE STERILE PHENOTYPE OF
MSG32 53
2.3.1 GENETIC MAPPING OF MSG32 53
2.3.2 IDENTIFICATION OF NEW MUTANT HVALDH2B ALLELES VIA CRISPR-CAS9 54
II
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/1206379111
TABLE OF CONTENTS
2.4 FUNCTIONAL CHARACTERIZATION OF MSG32 56
2.4.1 PHYLOGENETIC ANALYSIS 56
2.4.2 EXPRESSION ANALYSIS OF HVALDH2B 58
2.4.3 COMPARATIVE TRANSCRIPTOMICS OF WILD-TYPE AND MSG32 REPRODUCTIVE
TISSUES 59
CHAPTER II 64
2.5 PHENOTYPIC CHARACTERIZATION OF MSG36.BK 64
2.5.1 MORPHOLOGICAL DESCRIPTION OF MSG36.BK 64
2.5.2 HISTOLOGICAL FEATURES OF MSG36.BK ANTHERS 65
2.6 IDENTIFICATION OF THE MSG36 GENE 66
2.7 FUNCTIONAL CHARACTERIZATION OF MSG36 69
2.7.1 EXPRESSION ANALYSIS OF MSG36.BK 69
2.7.2 STUDYING THE LEVEL OF FUNCTIONAL CONSERVATION IN PECTIN-DEGRADING
ENZYMES 70
3. DISCUSSION 72
CHAPTER I 72
3.1 MSG32 IS INVOLVED IN POST-MEIOTIC MICROSPORE DEVELOPMENT AND TAPETUM
FUNCTION 72
3.1.1 MSG32 DEFECTS APPEAR POST-MEIOTICALLY UPON MICROSPORE VACUOLATION
AND ALTER MICROSPORE
DEVELOPMENT AND TAPETUM FUNCTION 72
3.1.2 MSG32 TAPETUM DEGENERATION INITIATED ON TIME BUT WAS IMPAIRED IN
AUTOPHAGY-RELATED
PROCESSES 76
3.1.3 POTENTIAL FUNCTION OF HVALDH2B IN ANTHER DEVELOPMENT OF BARLEY 77
3.1.4 CONCLUSIONS AND FUTURE PERSPECTIVES 82
CHAPTER II 84
3.2 MSG36 IS REQUIRED FOR CELL SEPARATION PROCESSES IN ANTHER OPENING 84
3.2.1 MSG36.BK DEFECTS OCCURRED IN SEPTUM AND STOMIUM CELLS UPON
MUTATION OF HVPL 84
3.2.2 PRELIMINARY EXPRESSION PROFILE OF HVPL AND STUDIES OF ARABIDOPSIS
HOMOLOGS 85
3.2.3 CONCLUSION AND FUTURE PERSPECTIVES 86
4. MATERIAL AND METHODS 87
4.1 PLANT MATERIAL 87
4.1.1 ARABIDOPSIS 87
4.1.2 BARLEY 87
4.2 PLANT CULTIVATION 88
4.2.1 CHAMBER CULTIVATION OF ARABIDOPSIS 88
4.2.2 GREENHOUSE CULTIVATION OF BARLEY 88
4.3 PHENOTYPING/SCORING OF DEVELOPMENT 89
4.4 COMPLEMENTATION TESTS OF BARLEY 89
4.5 ARABIDOPSIS CROSSING 90
4.6 ALEXANDER'S STAIN TO TEST POLLEN VIABILITY 90
4.7 SCANNING ELECTRON MICROSCOPY (SEM) 90
4.8 HISTOLOGICAL ANALYSIS 91
4.8.1 CHEMICALLY FIXED BARLEY FLOWERS 91
III
TABLE OF CONTENTS
4.8.2 BARLEY FLOWERS FIXED BY HIGH-PRESSURE FREEZING/FREEZE SUBSTITUTION
(HPF/FS) 91
4.9 ULTRASTRUCTURAL ANALYSIS OF CHEMICALLY FIXED AND HIGH-PRESSURE
FROZEN/FREEZE-SUBSTITUTED
BARLEY FLOWERS 92
4.9.1 CHEMICALLY FIXED BARLEY FLOWERS 92
4.9.2 BARLEY FLOWERS ANALYSED BY HPF/FS 93
4.10 TRANSMISSION ELECTRON MICROSCOPY (TEM) 94
4.11 POLLEN WALL MEASUREMENTS OF CHEMICALLY FIXED SAMPLES 94
4.12 DNA FRAGMENTATION DETECTION/TUNEL ASSAY 94
4.13 RNA IN SITU HYBRIDISATION OF BARLEY 96
4.14 COLLECTION OF PLANT MATERIAL FOR QRT-PCR AND TRANSCRIPTOME
SEQUENCING FOR DIFFERENTIALLY
EXPRESSED TRANSCRIPT (DET) ANALYSIS 97
4.15 TRANSCRIPTOME SEQUENCING FOR VARIANT CALLING FOR THE IDENTIFICATION
OF CANDIDATE GENES 98
4.16 RNA EXTRACTION, CDNA SYNTHESIS AND QRT-PCR 100
4.17 DNA EXTRACTION 101
4.18 MAPPING FOR CANDIDATE GENE IDENTIFICATION 102
4.19 OLIGONUCLEOTIDES 102
4.20 POLYMERASE CHAIN REACTIONS (PCR) 108
4.21 RESTRICTION ENDONUCLEASE DIGESTION AND AGAROSE GEL ELECTROPHORESIS
OF DNA 108
4.22 PLASMID CONSTRUCTION FOR GENERATION OF TRANSGENIC BARLEY PLANTS 109
4.22.1 CRISPR-CAS9 PLASMID CONSTRUCTION 109
4.22.2 PLANT TRANSFORMATION 110
4.23 SEQUENCE INFORMATION AND DISPLAY 111
4.24 IDENTIFICATION OF HOMOLOGS AND PHYLOGENETIC ANALYSIS 111
5. REFERENCES 114
6. SUPPLEMENTARY DATA 131
7. CURRICULUM VITAE 135
8. AFFLDAVIT/EIDESSTATTLICHE ERKLARUNG 137
9. ACKNOWLEDGEMENT 138
IV |
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| author_GND | (DE-588)1204850062 |
| author_facet | Przybyl, Marine 1989- |
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| spelling | Przybyl, Marine 1989- Verfasser (DE-588)1204850062 aut Characterization of genes involved in pollen development and anther opening in barley vorgelegt von Marine Przybyl Köln 2020 txt rdacontent n rdamedia nc rdacarrier Dissertation Universität zu Köln 2020 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1206379111/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032443892&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Przybyl, Marine 1989- Characterization of genes involved in pollen development and anther opening in barley |
| subject_GND | (DE-588)4113937-9 |
| title | Characterization of genes involved in pollen development and anther opening in barley |
| title_auth | Characterization of genes involved in pollen development and anther opening in barley |
| title_exact_search | Characterization of genes involved in pollen development and anther opening in barley |
| title_exact_search_txtP | Characterization of genes involved in pollen development and anther opening in barley |
| title_full | Characterization of genes involved in pollen development and anther opening in barley vorgelegt von Marine Przybyl |
| title_fullStr | Characterization of genes involved in pollen development and anther opening in barley vorgelegt von Marine Przybyl |
| title_full_unstemmed | Characterization of genes involved in pollen development and anther opening in barley vorgelegt von Marine Przybyl |
| title_short | Characterization of genes involved in pollen development and anther opening in barley |
| title_sort | characterization of genes involved in pollen development and anther opening in barley |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1206379111/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032443892&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT przybylmarine characterizationofgenesinvolvedinpollendevelopmentandantheropeninginbarley |
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