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Improvement of virus resistance breeding in barley by the help of H. bulbosum

Improvement of virus resistance breeding in barley by the help of H. bulbosum. Julia Kretsch, Dragan Perovic, Antje Habekuß , Viktor Korzun, Klaus Oldach , Neele Wendler, Hélène Pidon, Nils Stein, Frank Ordon. BYDV and BaMMV/BaYMV. Dr. Antje Habekuß , JKI. Dr. Antje Habekuß , JKI. 2.

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Improvement of virus resistance breeding in barley by the help of H. bulbosum

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  1. Improvement of virus resistance breeding in barley by the help of H. bulbosum • Julia Kretsch, Dragan Perovic, Antje Habekuß, Viktor Korzun, Klaus Oldach, Neele Wendler, Hélène Pidon, Nils Stein, Frank Ordon

  2. BYDV and BaMMV/BaYMV Dr. Antje Habekuß, JKI Dr. Antje Habekuß, JKI 2

  3. Resistance donor Hordeum bulbosum • Secondary genepool • Genetic potential for barley breeding • „New“ resistances and tolerances • But also negative traits and lower recombination rate I. Hordeum vulgare subsp. vulgareH. vulgare subsp. spontaneum II. Hordeum bulbosum III. ~ 30 Hordeum species Introgression lines • Small chromosome section of H. bulbosum was integrated into the genome of barley • Less linkage-drag caused by a reduction of the H. bulbosum fragment • But further reduction of the H. b. introgression is necessary Dr. Richard Pickering, The New Zealand Institute for Plant and Food Research Limited Dr. Paul A. Johnston, The New Zealand Institute for Plant and Food Research Limited 3

  4. Aim and workflow Identification and cloning of genes for resistance/tolerance against BYDV and BaMMV/BaYMV in Hordeum vulgare • Molecular and phenotypical testing of different DHand F2 populations • High-resolution mapping • Use ofGenotypingbySequencing (GBS), Exome Capture and Illumina 50K data • Resistance gene enrichment sequencing (RenSeq) • Screening of a non-gridded BAC library

  5. Previous work on Rym16Hb and Ryd203S11Hb Chr 2HL Chr 2HL Marker name Distance (cM) Marker name Distance (Mb) Rym16Hb Ryd203S11Hb 3 Mb Rym16Hb Perovic et al., unpublished Johnston et al., 2015 Ruge-Wehling et al., 2006

  6. Genotyping Workflow: Conversion of “old” flanking markers to KASP-markers for high-throughput analysis Molecular analysis of F2 and DH populations Detection of recombinant plants Resistant parent Susceptible parent Recombinant offspring Marker 1 Phenotyping Marker 2

  7. Phenotyping BaMMV / BaYMV for Rym16Hb • Field test on virus-contaminated field • Symptom scoring • ELISA • Climate chambertest • Mechanical virusinoculation • Symptom scoring • ELISA (only possible for BaMMV) • Phenotypical scoring: • Date of heading • Plant height • Yield parameters Dr. Antje Habekuß, JKI Dr. Antje Habekuß, JKI Performance of infected plant Performance of control plant = Tolerance in % 7 BYDV for Ryd203S11Hb • Test fortolerancein the gauze house

  8. ResultsofRym16Hb 4246 F2 analyzed plants 32 recombinant plants This represents 0.38 cMin an interval of 10 Mbp 16 of them were genotyped with an Illumina 50K chip 10 Mbp First F4 RILs will be phenotyped in autumn

  9. ResultsofRyd203S11Hb 12084 F2 seeds available ~ 8000 plants were analyzed 2 recombinant plants were found One of them was genotyped with the 50K chip RILs of this plant are currently in phenotyping

  10. RenSeq for Rym16Hb Resistance gene enrichment Sequencing No sequencing of the whole genome, but only sequencing of enriched DNA based on the information of resistance genes • General benefits • Usable in areas with low recombination rate • No fine-mapping necessary (but helpful) • Usable to get R genes from sexually incompatible species • de novo assembly of R genes of previously unsequenced, highly divergent wild relatives possible • Our plan • RenSeq of • H.b. donor lines (Rym16Hb) • Recombinant plants (susc. / res.) 10 Jupe et al., 2013

  11. BAC library for Ryd203S11Hb Ryd203S11Hb Interval Marker 1 BAC1 114 kb Non-gridded Bacterial Artificial Chromosome library BAC libraries are large DNA insert libraries of fragmented DNA of choice Marker 2 Marker 3 • The BAC library was screened with markers in the interval of Ryd203S11Hb to find BAC clones with DNA fragments of interest • These BACs were sequenced • The sequence will be analyzed for genes of interest (R genes) Marker 4 BAC2 109 kb Marker 5 4.2 Mbp • Usefulness of these BACs will become clear after phenotyping of RILs (to shorten the interval) Marker 6 Marker 7 Marker 8 Marker 9

  12. Outlook • Further geno- and phenotyping of populations for Ryd203S11Hband Rym16Hb • High-resolution mapping and marker saturation by help of Exome Capture, GBS and Illumina 50K data • Analyzing the non-gridded BAC-library for the tolerance Ryd203S11Hb • RenSeq for the resistance Rym16Hb • Mapbasedcloning The plant material and the diagnostic markers will accelerate the breeding of resistant cultivars.

  13. Thanksto: KWS LOCHOW GMBH (coordinator) Dr. Viktor Korzun / Dr. Jörg Großer / Anja Maasberg / Dr. Neele Wendler Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)Genomics of Genetic Resources (GGR) Prof. Dr. Nils Stein / Dr. Hélène Pidon Julius-Kühn Institute, Federal Research Centre for Cultivated Plants Institute for Resistance Research and Stress Tolerance Prof. Dr. Frank Ordon / Dr. Antje Habekuß / Dr. Dragan Perovic Technical assistance: Anne Kersten and Kerstin Herz Förderkennzeichen: 2818201515

  14. Thank you for your attention!

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