Aus dem Institut für Pflanzenzüchtung und Pflanzenschutz Advanced Backcross QTL analysis and genetic study of an introgressed powdery-mildew resistance gene derived from Avena macrostachya in oat (Avena sativa) Der Landwirtschaftlichen Fakultät der Martin-Luther-Universität Halle-Wittenberg als Dissertation zur Erlangung des akademischen Grades doctor agriculturarum (Dr. agr.) vorgelegt von M.Sc. in Bio. Jianzhong Yu geb. am 21.01.1976 in Hubei, Volksrepublik China Gutachter: Prof. Dr. W.E. Weber Prof. Dr. P. Wehling PD Dr. S. Maak Verteidigung am: 24.10.2005 Halle/Saale 2005 urn:nbn:de:gbv:3-000009169 [http://nbn-resolving.de/urn/resolver.pl?urn=nbn%3ade%3agbv%3a3-000009169]
Table of Contents Table of Contents... i Abbreviations... iv 1 Summary... 1 1.1 Oat AB-QTL analysis... 1 1.2 Genetic study of an introgressed powdery-mildew resistance gene derived from Avena macrostachya in oat (Avena sativa)... 2 2 Zusammenfassung... 4 2.1 AB-QTL-Analyse bei Hafer... 4 2.2 Genetische Charakterisierung der introgressierten Mehltauresistenz aus Avena macrostachya im Saathafer (Avena sativa)... 5 3 Literature review... 7 3.1 Oat: an important cereal crop... 7 3.1.1 Oat β-glucan: physiological values... 7 3.1.2 Oat β-glucan: contents and methods for analysis... 8 3.1.3 Oat β-glucan: inheritance and correlated responses... 8 3.2 Genetic markers... 9 3.2.1 Conventional genetic markers... 9 3.2.2 DNA-based molecular markers...11 3.3 Advanced backcross QTL analysis... 13 3.4 Oat genome and mapping... 15 3.5 QTL analysis in oat... 16 3.6 Oat powdery-mildew resistance... 18 3.6.1 Resources of oat powdery-mildew resistances... 18 3.6.2 Avena macrostachya, a useful wild species for oat breeding... 20 3.6.3 Application of molecular markers for disease resistance in oat.... 20 4 Materials and Methods... 22 4.1 AB-QTL analysis... 22 4.1.1 Population development... 22 4.1.2 Field evaluations... 22 4.1.3 Trait evaluations... 24 4.1.4 DNA isolation... 25 4.1.5 SSR analysis... 25 4.1.6 AFLP analysis... 26 4.1.7 Statistical analysis... 27 4.2 Genetic analysis of oat powdery-mildew resistance... 28 4.2.1 Plant materials... 28 4.2.2 Resistance testing... 30 4.2.3 DNA isolation and bulked segregant analysis... 30 i
4.2.4 SSR and AFLP analysis... 31 4.2.5 Cloning and conversion of AFLP markers... 31 4.2.6 Linkage analysis... 31 5 Results... 32 5.1 AB-QTL analysis... 32 5.1.1 Phenotypic traits... 32 5.1.2 ANOVA for genotypes and environments... 36 5.1.3 Correlation between traits... 38 5.1.4 Evaluation of agronomic values... 41 5.1.5 Marker polymorphism and segregation... 41 5.1.6 Map development... 41 5.1.7 QTL analysis... 43 5.2 Genetic analysis of oat powdery-mildew resistance... 50 5.2.1 Inheritance of the powdery-mildew resistance... 50 5.2.2 SSR polymorphisms and BSA analysis... 52 5.2.3 AFLP polymorphisms and BSA analysis... 53 5.2.4 Cloning and conversion of AFLP markers... 54 5.2.5 Linkage analysis... 57 6 Discussion... 59 6.1 AB-QTL analysis... 59 6.1.1 Phenotypic distribution... 59 6.1.2 Trait correlation... 60 6.1.3 ANOVA analysis... 62 6.1.4 Evaluation of agronomic values... 63 6.1.5 Marker polymorphism and segregation... 63 6.1.6 Map development... 64 6.1.7 QTL analysis... 66 6.1.7.1 Method for QTL analysis... 66 6.1.7.2 QTL analysis in advanced backcross population... 66 6.1.7.3 Conservation of QTLs across environments... 67 6.1.7.4 Co-localization of QTLs... 68 6.1.7.5 QTL comparison and validation... 68 6.2 Genetic analysis of oat powdery-mildew resistance... 69 6.2.1 Avena macrostachya is an important genetic resource for oat breeding... 69 6.2.2 Inheritance of powdery-mildew resistance... 70 6.2.3 BSA is powerful for development of linked markers... 70 6.2.4 SSR as molecular markers for MAS in oat... 71 6.2.5 AFLP mapping of Eg5 and AFLP-derived STS markers development... 71 6.2.6 Mapping of resistance gene Eg5... 73 7 Conclusions and outlooks... 75 7.1 Oat AB-QTL analysis... 75 ii
7.2 Genetic study of an introgressed powdery-mildew resistance gene derived from Avena macrostachya in oat (Avena sativa)... 76 8 References... 78 iii
Abbreviations Abbreviations 2D-PAGE AB AFLP ANOVA APR bp BSA BYDV CIM cm CTAB DNA DWD EDTA EST FIA HPLC LOD MAS NIL NIRS OMR PAGE PCR QTL RAPD RFLP RIL SIM SNP SPA SSR STS TE Tris USDA two-dimensional polyacrylamide gel electrophoresis advanced backcross amplified fragment length polymorphism analysis of variance adult plant resistance basepair bulked segregant analysis barley yellow dwarf virus composite interval mapping centimorgan cetyltrimethylammonium bromide deoxyribonucleic acid deutscher wetterdienst ethylenediaminetetraacetic acid expressed sequence tag flow injection analysis high performance liquid chromatography logarithm of odds marker assisted selection near isogenic line near infrared reflectance spectroscopy oat mildew resistance olyacrylamide gel electrophoresis polymerase chain reaction quantitative trait locus random amplified polymorphic DNA restriction fragment length polymorphism recombinant inbred line simple interval mapping single nucleotide polymorphism single point analysis simple sequence repeat sequence-tagged-site Tris/EDTA 2-Amino-2 (hydroxymethyl)-1,3-propandiol united states department of agriculture iv