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Prebreeding for resistance – examples from barley and wheat Frank Ordon www.jki.bund.de Prebreeding Effective Screening methods Identification of donors of resistance Genetics and marker development H. vulgare Cultivars Landraces H. vulgare ssp. spontaneum H. bulbosum Hordeum sp. n=30 Institute for Resistance Research and Stress Tolerance Marker based exploitation Marker assisted selection Marker assisted backcrossing Pyramiding etc. Barley yellow mosaic virus disease First discovery in Europe: 1978 Germany 1980 UK Cultivar BaYMV/BaMMVBaYMV/BaMMV- Yield Reaction t/ha relative Asorbia (6(6-rowed) Corona (6(6-rowed) Romanze (2(2-rowed) Marinka (2(2-rowed) resistant susceptible resistant susceptible 5.33 3.53 4.20 2.38 100 65 100 57 Yuka (6(6-rowed) Grete (6(6-rowed) Duet (2(2-rowed) Angora (2(2-rowed) resistant susceptible resistant suceptible 7.66 4.10 6.30 4.24 100 54 100 67 W. Huth 10 m2 plots, 3 replications; LSD (5%) = 0.25 t/ha and 0.35 t/ha; yield yield in ATM BaMMV, BAMMV-SIL, BaMMV-EIK, BaYMV, BaYMV-2 Institute for Resistance Research and Stress Tolerance Economic losses caused by BaMMV/BaYMV Acreage (2008): 1418200 ha Yield: 6.61 t = 9374302 t W. Huth Barley price: 140 € pro t Economic value: 1312402280 Euro 50% of barley acreage potentially infested (Huth 1988 ): 709100 ha Moderate yield loss of 25%: 1171787 t Economic loss: 164050180 € Institute for Resistance Research and Stress Tolerance Screening for resistance and genetic analysis BaMMV BaYMV BaYMV-2 No. Genotypes Hordeum vulgare ssp. vulgare 201 + 86 + 53 + + 43 Hordeum vulgare ssp. spontaneum + + 3 2 - = negative ELISA (resistant), + = positive ELISA ( susceptible) Ordon et al. 1993, Habekuss et al. 2000 F2 Segregation r : s χ2** 7r:9s H. vulgare ssp vulgare Chikurin Ibaraki 1 x Ogra Russia 57 x Diana Bulgarian 347 x Diana 49 : 69 68 : 81 61 : 89 0.233 0.214 0.574 98 95 95 Bulgarian 347 x Russia 57 Taihoku A x Chikurin Ibaraki 1 Bulgarian 347 x Chikurin Ibaraki 42 : 73 50 : 68 53 : 63 2.442 0.091 0.0114 100 100 100 Cross Institute for Resistance Research and Stress Tolerance Infect.rate (%) Götz & Friedt 1993, Ordon & Friedt 1993 Kartierung und Entwicklung molekularer Marker Rym14HB rym7 rym11 rym1 rym13 rym9 rym8 rym4 rym5 rym6 rym10 Rym16HB rym15 rym3 Chikurin Ibaraki 1 100cM 7H(1) 2H 3H 4H Distanz Marker cM name BaYMV/BaYMV-2 Resistenz Bmac0306 Bmac0163 0.0 Bmac0113 3.2 Ebmac0970 1.1 Bmag0394 5H Institute for Resistance Research and Stress Tolerance 1H(5) 6H 5H(7) mod. Graner et al. 2000, Ordon et al. 2005 Pyramiding of BaMMV/BaYMV Resistance Genes BaYMV BaYMV-2 BaMMV BaMMV-SIL BaMMV-EIK rym4 r s r r r rym5 r r r s s rym9 s s r r r rym11 r r r r r rym4 x rym9 x rym11 rym4 x rym9 rym4 rym4 x rym11 rym9 x rym11 rym5 Institute for Resistance Research and Stress Tolerance rym5 x rym9 x rym11 rym5 x rym9 rym9 rym5 x rym11 rym11 Pyramiding of BaMMV/BaYMV Resistance Genes 1 Locus homozygous Kreuzung P rym4 RYM9 rym4 RYM9 F1 F1´ X RYM4 rym9 rym4 RYM11 RYM4 rym9 rym4 RYM11 rym4 rym9 RYM11 RYM4 RYM9 RYM11 rym4 RYM9 RYM4 RYM11 rym4 RYM11 RYM9 RYM4 rym11 RYM9 RYM4 RYM9 rym4 rym11 RYM9 rym9 rym11 RYM4 rym11 rym4 rym11 RYM9 RYM4 RYM11 RYM9 RYM9 RYM4 RYM11 RYM4 RYM11 RYM9 RYM11 RYM4 rym9 RYM11 RYM4 RYM9 RYM11 rym4 rym9 RYM11 rym4 RYM9 RYM11 rym4 RYM9 RYM11 rym4 RYM9 RYM11 rym4 RYM9 RYM11 rym4 RYM9 RYM9 RYM11 rym11 RYM4 RYM4 rym9 RYM9 RYM11 rym11 RYM4 RYM9 RYM11 rym4 RYM4 RYM9 rym11 RYM4 rym9 RYM9 RYM11 RYM9 RYM11 RYM4 rym9 RYM11 RYM4 RYM9 RYM11 rym4 rym9 RYM11 RYM4 RYM9 RYM11 RYM4 RYM9 RYM11 RYM4 RYM9 RYM11 RYM4 RYM9 RYM11 rym4 RYM9 RYM11 RYM4 rym9 RYM11 RYM4 RYM9 RYM11 rym4 rym9 RYM11 rym4 RYM9 RYM9 rym11 RYM9 rym11 rym4 Bmac 29 (rym4) rym4 rym4 HVM 67 (rym9) rym9 x rym4 X rym9 x rym11 11 rym11 x rym4 X rym11 x rym9 3 rym5 x rym9 X rym5 x rym11 3 rym9 x rym5 X rym9 x rym11 3 rym11 x rym5 X rym11 x rym9 8 - RYM9 HVM 03 (rym11) 14 6 rym11 rym4 rym11 5 1 rym9 rym4 RYM11 rym4 x rym9 X rym4 x rym11 rym4 RYM4 RYM11 X x X x 2 Loci heterozygous 3 Loci heterozygous 8 - rym11 DH Line Production A1 rym4 rym9 rym1 1 rym4 rym9 RYM 11 rym4 RYM 9 rym1 1 rym4 RYM 9 RYM 11 rym4 rym9 rym1 1 rym4 rym9 RYM 11 rym4 RYM 9 rym1 1 rym4 RYM 9 RYM 11 Marker Selection Werner K, W Friedt, F Ordon, 2005. Mol. Breeding 16: 45-55 Institute for Resistance Research and Stress Tolerance rym4, rym9 and rym11 fixed rym4 and rym9 fixed rym4 and rym11 fixed Pyramiding of BaMMV/BaYMV Resistance Genes 4r 4r 4r 9r 4r 9r 9r 5r 5r 11r 11r 9r 11r 11r rym4 x rym9 x rym11 rym5 x rym9 x rym11 20 von 107 DHs, Chi2 = 0,072 27 von 187 DHs, Chi2 = 2,186 9r 9r 11 11 4r 4r 9 9 11r 11r 4/5 4/5 9r 9r 11r 11r 5r 5r 9 9 11r 11r 5r 5r 9r 9r 11 11 rym4 x rym9 rym4 x rym11 rym9 x rym11 rym5 x rym11 rym5 x rym9 31 von 107 DHs 17 von 107 DHs 42 von 294 DHs 21 von 187 DHs 31 von 187 DHs Chi2 =7,489 Chi2 =0,086 Chi2 =0,033 Chi2 =1,642 Chi2 =0,629 Werner K, W Friedt, F Ordon, 2005. Mol. Breeding 16: 45-55 Institute for Resistance Research and Stress Tolerance Gen Identification Rym4/Rym5 High resolution mapping A P11M36 0.10 P21M68 B MWG838 0.10 0.05 0.10 0.97 0.05 rym4 MWG838+ Y57C10+ E37*M36 E37*M63 E35*M81 E39*M39 STSE31*M41+ rym5 0.034 E62*M67 0.82 P20M46 0.05 B368N12 OP-Z04 0.34 0.88 OP-AF18H971 E45*M69 MWG010 BMac029 0.34 0.66 figure taken from: Buchanan, Gruissem, Jones (2000) Biochemistry & Molecular Biology of plants OP-AF18H971+ E45*M69 MWG010+ BMac029* Pellio et al. 2005 BAC-contig Co-segregation proximal 100 kb Institute for Resistance Research and Stress Tolerance eIF4E allele mining 1000 accessions selected year1 resequencing known haplotypes 40 novel exon haplotypes resistance tests 13 susceptible 27 resistant haplotypes year2/3 test crosses, resistance tests year3 Institute for Resistance Research and Stress Tolerance non allelic genes identification of 8 novel eIF4E alleles A. Graner Leaf rust (Puccinia hordei) in wheat Use of Triticum monococcum Lr37 2007 2005 Institute for Resistance Research and Stress Tolerance Accession 7038 7041 36594 37351 37353 BGRC 42007 BGRC 042013 BGRC 042017 EK 2 PI 167526 PI 272560 PI 352473 PI 352475 PI 355538 PI 428174 TRI 00620 TRI 17633 TRI 17668 TRI 577/81 TRI 645/76 TRI 1988/76 TRI 1989/76 TRI 1996/75 TRI 2125/91 TRI 4304/74 TRI 00620 PTR r r r r r r r r r r r mr r r mr r r Pow dery Mildew s r r r r r s s s s r r r r r r r r r r r r Leaf rust r r s s s r r r r r s r mr r r r r r r r r r T.monococcum (resistant) T. monococcum (medium susceptibility) Wheat (susceptible) Prehaustorial resistance 48 h post inoculation 96 h post inoculation 168 h post inoculation 10 d pst inoculation Balken = 20 µm A. Serfling Institute for Resistance Research and Stress Tolerance Preliminary results of QTL-analyses Institute for Resistance Research and Stress Tolerance Stem rust (Puccinia graminis) IRAN 2008 http://image.space.rakuten.co.jp/lg01/88/0000061588/25/imgb9 01d9fazikbzj.jpeg Institute for Resistance Research and Stress Tolerance Analyses of Pgt races Shewa 13 races (DPBTR, KHHTT, MLBDC, PTHSR, QLDGH, QMQTR, RKHSR, RRTTR, TCHTT, THHST, TTHSR, TTKSR, TTTTR) Arsi 8 races (KCCST, PTHSR, RMTTM, RRTTR, TTHSH, TTHSR, TTHTR, TTKSR) Bale 7 races (KCCST, MRHLR, PTHSR, RRTTR, TTHSR, TTKSR) NWE 8 races (HRTSH, KCCST, KRHST, QQQCM, RRTTR, TMHSR, TTHSR, TTKSR) Total 22 races . 2009 Admassu et al Institute for Resistance Research and Stress Tolerance Effectiveness of resistance genes Differential host Resistance gene Shewa Arsi NWEa Bale Total No. % No. % No. % No. % No. % CnSSr9g Sr9g 51 94.4 39 100 33 100 25 96.2 148 97.4 W2691Sr10 Sr10 53 98.1 39 100 31 93.9 25 96.2 148 97.4 ISr11-Ra Sr11 39 72.2 37 94.9 32 97 22 84.6 130 85.5 W2691Sr13 Sr13 10 18.5 2 5.1 1 3 9 34.6 22 14.5 Combination VII Sr17 48 88.9 39 100 33 100 25 96.2 145 95.4 CS_T_mono_deriv Sr21 43 79.6 34 87.2 27 81.8 26 100 130 85.5 Bt Sr30 Wst Sr30 21 38.9 19 48.7 17 51.5 9 34.6 66 43.4 W2691SrTt-2 Sr36 12 22.2 7 17.9 2 6.1 7 26.9 28 18.4 Triumph SrTmp 18 33.3 9 23.1 5 15.2 4 15.4 36 23.7 McNair 701 SrMcN 54 100 39 100 33 100 26 100 152 100 Admassu et al.2009 Institute for Resistance Research and Stress Tolerance Genetic Mapping of stem rust resistance gene Sr13 Sr13 – effective against Ug99 and other races Crossing between the susceptible cultivar (Morocco) and resistant line (Khapstein/9*LMPG ) 158 F2 population developed and phenotyped using well characterized Pgt isolate 10 – 15 progenies of each F2 plant re-tested A 0 XSSR1 11 XSSR2 15 XSSR3 18 XSr13 24 XSSR4 37 XSSR5 Admassu et al.(in prep.) Institute for Resistance Research and Stress Tolerance Pre-breeding in the future Puccinia triticina Einkorn (schwach anfällig) Weizen (anfällig) -Sequence information -omics Technologies Einkorn (resistent) Identification of genes and networks G. Hammer et al. 2006: Trends in Plant Science 11, 587-593 48 h nach Inokulation 96 h nach Inokulation Detection of allelic diversity 168 h nach Inokulation 10 d nach Inokulation mit Braunrost (Puccinia triticina) Balken = 20 µm Pre- breeding and breeding on the allele level Institute for Resistance Research and Stress Tolerance Prof. Dr. Andreas Graner Dr. Nils Stein Prof. Dr. Dr. h.c. W. Friedt Dr. Jens Weyen Dr. Andrea Schiemann Dr. Konstanze Scheurer Dr. Bettina Pellio Dr. Kay Werner Dr. Dragan Perovic Dr. Antje Habekuß Dr. Albrecht Serfling Dr. Volker Lind Belayneh Admassu DFG GFP BMBF BMELV KAAD EU Institute for Resistance Research and Stress Tolerance