Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana:
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| Format: | Thesis Book |
| Language: | English |
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2013
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| Online Access: | Inhaltsverzeichnis |
| Item Description: | Zsfassung in dt. und engl. Sprache |
| Physical Description: | IX, 114 S. Ill., graph. Darst. |
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| 100 | 1 | |a Glöckle, Barbara Magdalena |e Verfasser |0 (DE-588)1051427703 |4 aut | |
| 245 | 1 | 0 | |a Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana |c vorgelegt von Barbara Magdalena Glöckle |
| 264 | 1 | |c 2013 | |
| 300 | |a IX, 114 S. |b Ill., graph. Darst. | ||
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| 500 | |a Zsfassung in dt. und engl. Sprache | ||
| 502 | |a Köln, Univ., Diss., 2013 | ||
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| adam_text | Titel: Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana
Autor: Glöckle, Barbara Magdalena
Jahr: 2013
Contents
Contents.................................................................................................................................I
Kurzzusammenfassung.................................................................................................IV
Abstract...............................................................................................................................VI
Nomenclature..................................................................................................................VII
Figure and table index................................................................................................VIII
1 Introduction.................................................................................................................1
1.1 Plant life cycle...................................................................................................................1
1.1.1 Gametophyte development.................................................................................................2
1.2 Cell cycle.............................................................................................................................5
1.2.1 Cell-cycle regulation in plants............................................................................................6
1.2.1.1 CD Ks......................................................................................................................................................6
1.2.1.1 Cyclins..................................................................................................................................................8
1.2.1.2 E2F transcription factors...........................................................................................................10
1.3 Histones and Chromatin.............................................................................................12
1.3.1 Histone H3 in Arabidopsis thallana................................................................................13
1.3.2 Histone H3 methylation......................................................................................................14
1.3.3 Chromatin changes during male gametophyte development............................17
1.4 General outline and scope of this work.................................................................18
2 Results.........................................................................................................................20
2.1 Establishment of a protocol for whole mount immunostaining in pollen .20
2.1.1 Analysis of different histone marks...............................................................................21
2.1.2 Dynamic pattern of H3K27mel during male gametogenesis.............................23
2.1.3 Analysis of histone marks in chromatin mutants....................................................25
2.1.3.1 Characterization of the htrlO mutant line..........................................................................29
2.2 Analysis of single-celled pollen of cdka;l*/- fbll 7+/- and e2fa/fbll 7*1-
double mutants.........................................................................................................................31
2.2.1 Viability test of pollen from cdka;l*/- fbll7*7 and e2fa/- fbll7*/- mutants ....31
2.2.2 Single-celled pollen of cdka;l*/-ßll 7*/- expresses the vegetative cell marker
PR0csdl4- GUS............................................................................................................................................32
2.2.3 Germline marker PROhtrw:HTR10-RFP is absent in single-cell pollen of
cdka;l*/ßil7*/- plants........................................................................................................................33
2.2.4 H3K27mel immunostaining in single-celled pollen confirms vegetative
development of single-cell pollen..................................................................................................34
2.2.5 The single-celled pollen of cdka;l*/ßjll7*7- plants does not express
PR0htri2:HTR12-GFP............................................................................................................................36
2.2.6 Transposons become expressed in single-cell pollen............................................39
2.2.7 Single-celled pollen can germinate in vitro................................................................40
2.3 Combination of D-type cyclin (CYCD) mutations with cdka;l*l· orßll 7*7-41
2.3.1 D-type cyclins expressed in pollen.................................................................................41
2.3.2 Characterization of CYCD1;1 and CYCD2;1 T-DNA mutants................................43
2.3.3 Expression of D-type Cyclin in the corresponding T-DNA mutant...................44
2.3.4 D-type cyclin mutations in a cdka;!*/¦ or fbll7*l- background enhance the
mutant pollen phenotype..................................................................................................................45
2.4 Combination of the MULTICOPYSUPRESSOR OF IRAI (MSI1) mutation with
editor;!*/-.......................................................................................................................................48
2.5 Screening for mutants with altered segregation ratios...................................49
2.5.1 Characterization of two T-DNA insertion lines for CSLD1...................................52
2.5.2 The mutant allele of csldl/+ plants is not transmitted paternally...................53
2.5.3 Analysis of pollen from csldl+Y mutant plants..........................................................54
3 Discussion..................................................................................................................59
3.1 Regulation of cell-cycle progression.......................................................................59
3.2 Cell-cycle progression and chromatin remodeling............................................60
3.3 Cell-fate and pollen function.....................................................................................64
4 Materials and Methods..........................................................................................69
4.1 Plant work.......................................................................................................................69
4.1.1 Plant strains.............................................................................................................................69
4.1.2 Antibiotics and herbicide used for plant selection..................................................71
4.1.3 Seed sterilization....................................................................................................................71
4.1.3.1 Liquid seed sterilization.............................................................................................................71
4.1.3.2 Seed sterilization with chlorine gas......................................................................................72
4.1.4 Growing conditions...............................................................................................................72
4.1.5 Crossing......................................................................................................................................72
4.1.6 Pollen germination assay...................................................................................................73
4.1.7 Callose staining of in vitro germinated pollen tubes..............................................74
4.1.8 Whole mount immunohistology in pollen..................................................................74
4.1.8.1 Flower preparation......................................................................................................................74
4.1.8.2 Antibody staining..........................................................................................................................75
4.1.9 Antibodies.................................................................................................................................77
4.1.10 Viability staining of pollen..............................................................................................77
4.1.11 DAPI staining of pollen and fluorescence marker investigation....................78
4.1.12 GUS staining of pollen.......................................................................................................78
4.2 DNA work.........................................................................................................................79
4.2.1 DNA isolation of plants........................................................................................................79
4.2.1 PCR...............................................................................................................................................80
4.2.1.1 Genotyping PCR.............................................................................................................................80
4.2.1.2 Endpoint PCR with cDNA...........................................................................................................81
4.2.2 Gel electrophoresis...............................................................................................................81
4.2.3 qRT-PCR.....................................................................................................................................82
4.2.3.1 RNA extraction...............................................................................................................................82
4.2.3.2 cDNA preparation.........................................................................................................................82
4.2.3.3 Quantitative Real Time PCR......................................................................................................83
4.2.4 Primer and primer combinations...................................................................................84
Abbreviations and gene names..................................................................................91
Amino acid abbreviation..............................................................................................94
References........................................................................................................................95
Publication......................................................................................................................110
Acknowledgement........................................................................................................Ill
Erklärung........................................................................................................................HI
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| spelling | Glöckle, Barbara Magdalena Verfasser (DE-588)1051427703 aut Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana vorgelegt von Barbara Magdalena Glöckle 2013 IX, 114 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. und engl. Sprache Köln, Univ., Diss., 2013 (DE-588)4113937-9 Hochschulschrift gnd-content HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027469843&sequence=000004&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Glöckle, Barbara Magdalena Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana |
| subject_GND | (DE-588)4113937-9 |
| title | Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana |
| title_auth | Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana |
| title_exact_search | Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana |
| title_full | Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana vorgelegt von Barbara Magdalena Glöckle |
| title_fullStr | Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana vorgelegt von Barbara Magdalena Glöckle |
| title_full_unstemmed | Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana vorgelegt von Barbara Magdalena Glöckle |
| title_short | Dissection of pollen development and function using cell-cycle mutants in Arabidopsis thaliana |
| title_sort | dissection of pollen development and function using cell cycle mutants in arabidopsis thaliana |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027469843&sequence=000004&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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