6th Africa Agriculture Science Week and FARA General Assembly FINALS OF THE 3rd AFRICA-WIDE WOMEN AND YOUNG PROFESSIONALS IN SCIENCE COMPETITIONS 15-16 July 2013 6th Africa Agriculture Science Week and FARA General Assembly Accra, Ghana QTL mapping for resistance to race Ug99 and identification of stem rust resistance genes in Ethiopian durum wheat varieties Jemanesh Haile, Ayele Badebo, Miloudi Nachit and Marion Roder

Outline 1. Introduction 2. QTL mapping for resistance to race Ug99 Identification of stem rust resistance genes Conclusions and potential impact Dissemination strategies e Ethiopian Institute of Agricultural Research

Introduction Wheat World's major crop Ethiopia: Second largest producer of wheat in sub-Saharan Africa Third most grown crop World’s secondary centres of diversity Low productivity: < 2 t/ha e Ethiopian Institute of Agricultural Research

Introduction cont‘d Stem Rust Puccinia graminis f. sp. tritici (Pgt) Why is stem rust important? 70% yield loss Race Ug99 What is Ug99? Sr31 High virulence diversity and evolution rate e Ethiopian Institute of Agricultural Research

Ug99 race group is now in 11 countries Potential area under the risk from Ug99: 50 million ha of wheat 1 billion people will be affected 117 million tons production loss (ca. 19% world’s production) • Value US$16.4 billion (using US$140/t)

1. QTL mapping of resistance to Ug99 race of Puccinia graminis f. sp 1. QTL mapping of resistance to Ug99 race of Puccinia graminis f. sp. tritici Objective: Identification of QTL regions conferring resistance to race Ug99 e Ethiopian Institute of Agricultural Research

Plant materials: 95 RILs - F8 Parents: ‘Kristal’ × ‘Sebatel’ - ICARDA Material and Methods Plant materials: 95 RILs - F8 Parents: ‘Kristal’ × ‘Sebatel’ - ICARDA Experimental sites: Debre-Zeit and Denbi, Ethiopia between 2008-10 e Ethiopian Institute of Agricultural Research

Material and Methods cont‘d Genotyping: Genomic DNA Polymerase Chain Reaction (PCR) Amplifications of markers Fragment detection Genetic map construction: 209 SSR markers Mapmaker v 2.0 QTL analysis: CIM options of QTL Cartographer v 2.5 QSr.ipk (International Rules of Genetic Nomenclature) e Ethiopian Institute of Agricultural Research

Results of QTL Analysis Kaur et al . 2009 R2 = 6.0 Sr2 Xcfa2193 R2 = 5.0 XBF485004 XBE423242 Sr35 Xcfa2076 Xcfa2170 Xgwm480 QTL for resistance to stem rust Markers linked with reported major Sr genes Approximate position of centromere 10 e Ethiopian Institute of Agricultural Research

Results of QTL analysis cont‘d Xwmc580 Xdupw167 Xbarc104 Sr13 Xcfa2123 Xbarc121 Sr22 R2 = 5.4 R2 = 5.1 R2 = 9.3 R2 = 7.3 R2 = 12.6 QTL for resistance to stem rust Markers linked with reported major Sr genes Approximate position of centromere 10 e Ethiopian Institute of Agricultural Research

New QTL for Ug99 race resistance in durum wheat Summary Nine QTL regions confer resistance to Ug99 race were identified. QTL on 1AL (QSr.ipk-1A), 2AS (QSr.ipk-2A), 4BL (QSr.ipk-4B), 7A (QSr.ipk-7A.1) and 7BL (QSr.ipk-7B) have not been previously detected. New QTL for Ug99 race resistance in durum wheat Two QTL co-localized with chromosomal regions harboring known Sr genes Sr2 (QSr.ipk-3B), Sr22 (QSr.ipk-7A.2) e Ethiopian Institute of Agricultural Research

2. Haplotype analysis of molecular markers linked to stem rust resistance genes Objective: Identification of stem rust resistance genes in Ethiopian durum wheat varieties e Ethiopian Institute of Agricultural Research

Material and Methods Plant materials: 22 Ethiopia 4 ICARDA 26 Durum wheat varieties: 22 Ethiopia 4 ICARDA e Ethiopian Institute of Agricultural Research

Material and Methods Cont’d Marker analyses: 17 PCR markers Sr2, Sr13, Sr22 and Sr35 Check lines: Sr2 – Kingbird Sr13 – W2691SR13 Sr22 – SWSR22TB Sr35 – W3763-SR35 Data analysis: Haplotypes sorted - size of fragments Compare to check lines e Ethiopian Institute of Agricultural Research

Results of Haplotype Analysis Sr2 Variety GWM533 BARC133 KINGBIRD 120 122 Sebatel Hitosa e Ethiopian Institute of Agricultural Research

Results of Haplotype Analysis cont‘d Sr13 Variety BE403950 DUPW167 BARC104b WMC580 BARC104c W2691SR13 691 243 273 316 175 Sebatel Quamy Boohai 177 Cocorit-71 290 Cham-1 e Ethiopian Institute of Agricultural Research

Sr22 Sr35 – no variety Results of Haplotype Analysis cont‘d Variety CFA2019 CFA2123 WMC633 BARC121 SWSR22TB 168 234 119 170 Sebatel 249 223 228 Boohai 259 233 238 Mamouri 240 JennahKhetifa 253 227 Sr35 – no variety e Ethiopian Institute of Agricultural Research

only a limited number of Sr genes with diagnostic markers. Summary Resistance against Ug99 in ‘Sebatel‘ might be due to the combination of Sr2 and Sr22. It is likely that the tested varieties also had resistant genes which were not detected. only a limited number of Sr genes with diagnostic markers. e Ethiopian Institute of Agricultural Research

Conclusions and Potential Impact Markers closely linked to the QTL could be used for MAS for resistance to race Ug99. `Sebatel` could be used as a donor variety. The durum wheat varieties released in Ethiopia do not carry diverse Sr genes. Need to broaden the genetic base of the varieties. e Ethiopian Institute of Agricultural Research

Dissemination Strategies International Journals Haile et al. (2012) Mol. Breed. 30:1479-1493 Haile et al. (2013) Genet Resour Crop Evol. 60: 853-864 Posters 8th Internationâl Wheat Conference - Russia 10th Sequence-informed Crop Research Conference – Germany 2011 Field Crop Rust Symposium – USA 2012 Borlaug Global Rust Imitative Workshop – China Presentations 6th Plant Science Student Conference – Germany 2011 International wheat pathology training – Mexico 2013 National Wheat Review - Ethiopia Newsletters Ann. Wheat Newsl. 57:12-15 Ann. Wheat Newsl. 58:68-71

Thanks for your attention! ACKNOWLEDGEMENTS CTA, FARA, IFS, ANAFE, RUFORUM, NPCA, AGRA EIAR, ICARDA, IPK, Kassel University, German Academic Exchange Service Thanks for your attention! DAAD

It is not just about productivity and economics, it is now an issue of world food security