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Publié par | gottfried_wilhelm_leibniz_universitat_hannover |
Publié le | 01 janvier 2007 |
Nombre de lectures | 112 |
Langue | English |
Poids de l'ouvrage | 2 Mo |
Extrait
Characterisation and mapping of bacterial wilt (Ralstonia
solanacearum) resistance in the tomato (Solanum lycopersicum)
cultivar Hawaii 7996 and wild tomato germplasm
Von der Naturwissenschaftlichen Fakultät der
Gottfried Wilhelm Leibniz Universität Hannover
zur Erlangung des akademischen Grades eines
Doktorin der Gartenbauwissenschaften
-Dr. rer. hort.-
genehmigte Dissertation
von
Truong Thi Hong Hai, Master of Agriculture
geboren am 18. Juni 1976 in Nghe An, Vietnam
2007
Referentin: Koreferentin:
PD Dr. Elisabeth Esch Prof. Dr. Kerstin Wydra
Tag der Promotion: 14.12.2007
Table of contents i
TABLE OF CONTENTS
TABLE OF CONTENTS........................................................................................................i
LIST OF TABLES.................................................................................................................v
LIST OF FIGURES ..............................................................................................................vi
LIST OF APPENDIX TABLES………………………………………………………… viii
ABBREVIATIONSix
ABSTRACT...........................................................................................................................1
ZUSAMMENFASSUNG ......................................................................................................3
GENERAL INTRODUCTION..............................................................................................5
Chapter 1 Construction of a genetic linkage map for mapping bacterial wilt
resistance in the tomato cultivar Hawaii 7996
1.1 INTRODUCTION .....................................................................................................7
1.2 MATERIALS AND METHODS ............................................................................11
1.2.1 Plant materials.................................................................................................11
1.2.2 DNA preparation and quantification...............................................................11
1.2.2.1 DNA preparation.................................................................................11
1.2.2.2 DNA quantification.............................................................................12
1.2.3 DNA marker analysis......................................................................................13
1.2.3.1 AFLP analysis.....................................................................................13
1.2.3.2 Microsatellite or SSR analysis............................................................16
1.2.3.3 SNP analysis .......................................................................................18
1.2.3.4 Providing of DArT and RFLP markers...............................................20
1.2.3.5 Marker codes.......................................................................................20
1.2.3.6 Linkage analysis .................................................................................20
1.3 RESULTS ................................................................................................................21
1.3.1 Polymorphism screening between H7996 and WVa700 ................................21
1.3.2 Segregation analysis of polymorphic markers................................................23
1.3.3 Genetic linkage map of H7996 x WVa700.....................................................26
1.4 DISCUSSION..........................................................................................................34
1.4.1 Polymorphism between H7996 and WVa70034 Table of contents ii
1.4.2 Segregation distortion.....................................................................................34
1.4.3 Map construction ............................................................................................35
1.5 SUMMARY.............................................................................................................38
Chapter 2 Detection of QTLs for bacterial wilt resistance in Hawaii 7996 and its
relationship with morphological traits
2.1 INTRODUCTION ...................................................................................................39
2.2 MATERIALS AND METHODS ............................................................................43
2.2.1 Plant materials.................................................................................................43
2.2.2 Evaluation of resistance to bacterial wilt........................................................43
2.2.2.1 Bacterial strains and inoculation.........................................................43
2.2.2.2 Evaluation based on visual symptoms................................................44
2.2.2.3 Evaluation based on colonization degree............................................45
2.2.3 Evaluation of morphological traits .................................................................47
2.2.3.1 Experimental design ...........................................................................47
2.2.3.2 Sampling and data collection..............................................................47
2.2.4 Data analysis...................................................................................................49
2.2.5 QTL analysis49
2.2.6 Fine mapping ..................................................................................................50
2.2.6.1 Bulk segregant analysis ......................................................................50
2.2.6.2 Conversion of AFLP, DArT and RFLP markers into PCR-based
markers................................................................................................50
2.2.6.3 Inverse PCR ........................................................................................55
2.2.6.4 Randomly amplified microsatellite polymorphism (RAMP) .............56
2.3 RESULTS ................................................................................................................58
2.3.1 Resistance to strain Pss4 and Pss186 in F RILs ............................................58 9
2.3.2 Colonization by the pathogen in F RILs........................................................62 9
2.3.2.1 Protocol development .........................................................................62
2.3.2.2 Colonization by strain Pss4 in F RILs...............................................64 9
2.3.3 Morphological trait distribution......................................................................65
2.3.3.1 Sympodial index (SPI)........................................................................65
2.3.3.2 Fruit weight.........................................................................................65
2.3.3.3 Skin color............................................................................................67
2.3.3.4 Fruit quality.........................................................................................68 Table of contents iii
2.3.4 Correlation among traits .................................................................................70
2.3.5 QTL detection.................................................................................................73
2.3.5.1 QTLs linked to bacterial wilt resistance .............................................73
2.3.5.2 QTLs affecting morphological traits...................................................78
2.3.5.3 Single marker analysis........................................................................79
2.3.6 Fine mapping ..................................................................................................81
2.3.6.1 Bulk segregant analysis ......................................................................81
2.3.6.2 Conversion of AFLP, DArT and RFLP markers into PCR-based
marker form ........................................................................................81
2.4 DISCUSSION..........................................................................................................89
2.4.1 Resistance to bacterial wilt in H7996 and its associated QTLs......................89
2.4.1.1 Common QTLs important for resistance against race 1 strains..........89
2.4.1.2 Colonization by Pss4 and resistance to bacterial wilt in H7996.........91
2.4.1.3 Plausible strain-specific QTLs to race 1 strains..................................91
2.4.1.4 Plausible environment-specific QTLs to race 1 strains ......................92
2.4.1.5 Comparison of QTLs associated with resistance to race 1 and 3 strains
............................................................................................................92
2.4.2 Morphological traits and their associated QTLs..................