Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana:
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| Format: | Thesis Book |
| Language: | English |
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2011
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | VIII, 86 S. Ill., graph. Darst. |
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| adam_text | Titel: Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana
Autor: Kumar, Manoj
Jahr: 2011
Contents
CONTENTS
Contents I
Abstract IV
Zusammenfassung V
Publications VI
Abbreviations and gene names VII
1 Introduction.....................................................................................................1
1.1 Plant life cycle and seed formation.................................................................................................1
1.2 Endosperm development....................................................................................................................2
1.2.1 Syncytial phase....................................................................................................................................2
1.2.2 Cellular phase........................................................................................................2
1.2.3 Maturation phase...............................................................................................................................4
1.3 Genetic studies of endosperm development and seed growth...........................................4
1.4 Cell cycle regulators during endosperm development..........................................................5
1.5 Cell cycle control in plants..................................................................................................................6
1.5.1 Cyclin-Dependent Kinases...............................................................................................................8
1.5.2 Regulation of CDK-cyclin activity................................................................................................9
1.5.3 Cyclins....................................................................................................................................................10
1.5.3.1 Cyclin domains....................................................................................................................................12
1.6 Cyclins in Arabidopsis.......................................................................................................................12
1.6.1 B type cyclins......................................................................................................................................13
1.7 CYCB1 gene family in Arabidopsis...........................................................15
2 Aim of the study.............................................................................................17
3 Results............................................................................................................18
3.1 Expression and localisation analysis of CyclinBl genes in Arabidopsis......................18
3.1.1 Expression analysis of CyclinBl genes in Arabidopsis.....................................................18
3.1.2 localisation ofBl type cyclins in mitotic cells...................................................................21
3.2 Isolation of CyclinBl mutants........................................................................................................23
Contents
3.3 Analysis of CYCB1 mutants..............................................................................................................25
3.3.1 CYCB1 T-DNA mutants are null allele.....................................................................................25
3.3.2 CYCB1 single mutants show altered endosperm nuclei proliferation.......................26
3.3.3 Certain double mutant combinations ofCYCBls cause seed abortion.....................27
3.3.4 Endosperm nuclei proliferation defect in CYCB1 double mutants..............................30
3.3.5 cycbl;2-/- shows maternal effect in endosperm proliferation.....................................33
3.3.6 The cycbl;l-/-cycbl;2-/- and cycbl;2-/-cycbl;3-/- plants show sporophyte
defects....................................................................................................................................................34
3.3.7 Growth rate defect in primary roots of cycbl;l-/-cycbl;2-/-.......................................37
3.3.8 cycbl;l-/-cycbl;2-/- roots show mitotic defects................................................................38
3.3.9 Mitotic defects in cycbl.l-/-cycbl;2-/- endosperm..........................................................38
3.3.10 Ploidy analysis reveals the accumulation of cells in G2 phase...................................40
3.3.11 Marker lines confirm mitotic arrest of cells in G2-M in double mutants..............42
3.3.12 Stomata development remains unaltered in cycbl;l-/- cycbl;2-/-........................43
3.4 The seed abortion phenotype of cycbl;l and cycbl;2 double mutants is
significantly enhanced by additional loss of CYCB1;3..........................................................43
3.5 Analysis of quadruple mutants of CYCBljl, CYCB1;2, CYCB1;3 and CYCB1;4.............45
3.6 EDE1. a possible substrate for CYCB1/CDK complex?........................................................45
3.7 QTL analysis of seed weight using three sets of R1L populations...................................47
3.8 QTL analysis of autonomous endosperm development upon pollination with
cdka;l pollen..........................................................................................................................................48
4 Discussion.......................................................................................................51
4.1 Bl-type cyclins accumulate in G2/M and localise to chromatin and spindle during
mitosis......................................................................................................................................................51
4.2 cycbl;2-/- represents a maternal effect mutant.....................................................................53
4.3 CYCBls are required for nuclear division of endosperm and seed development in
Arabidopsis.............................................................................................................................................54
4.4 Embryo development in spite of compromised endosperm proliferation in cycbl;l-
/-cycbl;2-/- and cycbl;2-/-cycbl;3-/- seeds-----------------..................................................55
4.4.1 The endosperm fail to cellularise in cycbl;l-/-cycbl;2-/- and cycbl;2-/-cycbl;3-/-
seeds............................................................................................................................................................56
4.4.2 The cycbl;l-/-cycbl;2-/- and cycbl;2-/-cycbl;3-/- show sporophyte growth
defects..................................................~....................................................................................................57
4.4.3 The cycbl;l-/-cycbl;2-/- and cycbl;2-/-cycbl;3-/- show G2/M arrest...................57
Contents
4.4.4 The mitotic defects in cycbl;l-/-cycbl;2-/- roots..............................................................58
4.4.5 The mitotic defects in cycbl;l-/-cycbl;2-/- endosperm.................................................59
4.4.6 The triple mutants ofCYCBl;l, CYCB1;2 and CYCB1;3 indicategametophytic
defect.........................................................................................................................................60
4.5 The quadruple mutants of CYCB1;1, CYCB1;2, CYCB1;3 and CYCB1;4 cause
gametophytic defect...........................................................................................................................60
4.6 Identification of potential substrates for CYCB1/CDK complex?...................................61
4.7 Identification of QTL loci regulating seed mass.....................................................................62
4.8 Identification of one locus controlling the autonomous endosperm division upon
pollination with cdka;l pollen........................................................................................................64
5 Materials Methods......................................................................................65
5.1 Materials..................................................................................................................................................65
5.1.1 Chemicals and antibiotics.............................................................................................................65
5.1.2 Enzymes, primers and kits............................................................................................................65
5.1.3 Plant material....................................................................................................................................65
5.2 Methods...................................................................................................................................................66
5.2.1 Plant growth conditions................................................................................................................66
5.2.2 Genomic DNA preparation from plant tissue.......................................................................66
5.2.3 Identification of mutant alleles by PCR..................................................................................67
5.2.4 Quantitative PCR..............................................................................................................................68
5.2.5 Crossing of plants.............................................................................................................................69
5.2.6 Whole-Mount preparation of seeds..........................................................................................69
5.2.7 Stomata staining..............................................................................................................................69
5.2.8 Flow cytometry for ploidy analysis...............................................................................70
5.2.9 Feulgen staining................................................................................70
5.2.10 lmmunostaining.............................................................................................................................71
5.2.11 QTL mapping...................................................................................................................................72
6 References......................................................................................................73
Appendix.....................................................................84
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| spelling | Kumar, Manoj 1980- Verfasser (DE-588)1015524559 aut Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana vorgelegt von Manoj Kumar 2011 VIII, 86 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Köln, Univ., Diss., 2011 (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=024599578&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Kumar, Manoj 1980- Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana |
| subject_GND | (DE-588)4113937-9 |
| title | Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana |
| title_auth | Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana |
| title_exact_search | Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana |
| title_full | Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana vorgelegt von Manoj Kumar |
| title_fullStr | Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana vorgelegt von Manoj Kumar |
| title_full_unstemmed | Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana vorgelegt von Manoj Kumar |
| title_short | Analysis of cell proliferation and growth control during seed development in Arabidopsis thaliana |
| title_sort | analysis of cell proliferation and growth control during seed development in arabidopsis thaliana |
| topic_facet | Hochschulschrift |
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