Abstract
Two of the domains most widely shared among R genes are the nucleotide binding site (NBS) and protein kinase (PK) domains. The present study describes and maps a number of new oat resistance gene analogues (RGAs) with two purposes in mind: (1) to identify genetic regions that contain R genes and (2) to determine whether RGAs can be used as molecular markers for qualitative loci and for QTLs affording resistance to Puccinia coronata. Such genes have been mapped in the diploid A. strigosa × A. wiestii (Asw map) and the hexaploid MN841801-1 × Noble-2 (MN map). Genomic and cDNA NBS-RGA probes from oat, barley and wheat were used to produce RFLPs and to obtain markers by motif-directed profiling based on the NBS (NBS profiling) and PK (PK profiling) domains. The efficiency of primers used in NBS/PK profiling to amplify RGA fragments was assessed by sequencing individual marker bands derived from genomic and cDNA fragments. The positions of 184 markers were identified in the Asw map, while those for 99 were identified in the MN map. Large numbers of NBS and PK profiling markers were found in clusters across different linkage groups, with the PK profiling markers more evenly distributed. The location of markers throughout the genetic maps and the composition of marker clusters indicate that NBS- and PK-based markers cover partly complementary regions of oat genomes. Markers of the different classes obtained were found associated with the two resistance loci, PcA and R-284B-2, mapped on Asw, and with five out of eight QTLs for partial resistance in the MN map. 53 RGA-RFLPs and 187 NBS/PK profiling markers were also mapped on the hexaploid map A. byzantina cv. Kanota × A. sativa cv. Ogle. Significant co-localization was seen between the RGA markers in the KO map and other markers closely linked to resistance loci, such as those for P. coronata and barley yellow dwarf virus (Bydv) that were previously mapped in other segregating populations.


Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Acevedo M, Jackson EW, Chong J, Rines W, Harrison S, Bonman JM (2010) Identification and validation of quantitative trait loci for partial resistance to crown rust in oat. Genet Resist 100:511–521
Bakker E, Borm T, Prins P, van der Vossen E, Uenk G, Arens M, de Boer J, van Eck H, Muskens M, Vossen J, van der Linden G, van Ham R, Klein-Lankhorst R, Visser R, Smanta G, Bakker J, Goverse A (2011) A genomic-wide genetic map of NB-LRR disease resistance loci in potato. Theor Appl Genet 123:493–508
Barbosa-Neto JF, Siripoonwiwat W, O’Donoughue LS, Gray SM, Smith DM, Kolb FL, Gourmet C, Brown CM, Sorrells ME (2000) Chromosomal regions associated with yellow dwarf virus resistance in oat. Euphytica 114:67–76
Biruma M, Martin T, Fridborg I, Okori P, Dixelieus C (2012) Two loci in sorghum with NB-LRR encoding genes confer resistance to Colletotrichum sublineolum. Theor Appl Genet 124:1005–1015
Brueggeman R, Rostoks N, Kudrna D, Kilian A, Han F, Chen J, Druka A, Steffenson B, Kleinhofs A (2002) The barley stem rust-resistance gene Rpg1 is a novel disease-resistance gene with homology to receptor kinases. Proc Natl Acad Sci USA 99:9328–9333
Brugmans B, Wouters D, van Os H, Hutten R, van der Linden CG, Visser RGF, van Eck HJ, van der Vossen EAG (2008) Genetic mapping and transcription analyses of resistance gene loci in potato using NBS profiling. Theor Appl Genet 117:1379–1388
Bush AL, Wise RP (1996) Crown rust resistance loci on linkage groups 4 and 13 in cultivated oat. J Hered 87:427–432
Calenge F, van der Linden CG, Van de Weg E, Schouten HJ, Van Arkel G, Denance C, Durel CE (2005) Resistance gene analogues identified through the NBS-profiling method map close to major genes and QTL for disease resistance in apple. Theor Appl Genet 110:660–668
Chen G, Chong J, Prashar S, Procunier JD (2007) Discovery and genotyping of high-throughput SNP markers for crown rust resistance gene Pc94 in cultivated oat. Plant Breed 126:379–384
Cheng DW, Armtrong KC, Tinker N, Wight CP, He S, Lybaert A, Fedak G, Molnar SJ (2002) Genetic and physical mapping of Lrk10-like receptor kinase sequences in hexaploid oat (Avena sativa L.). Genome 45:100–109
Fernández-Silva I, Eduardo I, Blanca J, Esteras C, Picó B, Nuez F, Arús P, García-Mas J, Monforte AJ (2008) Bin mapping of genomic and EST-derived SSRs in melon (Cucumis melo L.). Theor Appl Genet 118:139–150
González AM, Marcel TC, Kohutova Z, Stam P, van der Linden CG, Niks RE (2010) Peroxidase profiling reveals genetic linkage between peroxidase gene clusters and basal host and non-host resistance to rusts and mildew in barley. PLoS ONE 5(8):e10495. doi:10.1371/journal.pone.0010495
Groh S, Zacharias A, Kianian SF, Penner GA, Chong J, Rines HW, Phillips RL (2001) Comparative AFLP mapping in two hexaploid oat populations. Theor Appl Genet 102:876–884
Grube RC, Radwanski ER, Jahn M (2000) Comparative genetics of disease resistance within the Solanaceae. Genetics 155:873–887
Gutierrez-Gonzalez JJ, Garvin DF (2011) Reference genome-directed resolution of homologous and homoelogous relationships within and between different oat linkage maps. Plant Genome 4:178–190
Howard W, Yamamoto T, Dirlewanger E, Testolin R, Cosson P, Cipriani G, Monforte AJ, Georgi L, Abbott AG, Arús P (2005) Mapping with a few plants: using selective mapping for microsatellite saturation of the Prunus reference map. Genetics 171:1305–1309
Irigoyen ML, Loarce Y, Fominaya A, Ferrer E (2004) Isolation and mapping of resistance gene analogs from the Avena strigosa genome. Theor Appl Genet 109:713–724
Jackson EW, Obert DE, Menz M, Hu G, Bonman JM (2008) Qualitative and quantitative trait loci conditioning resistance to Puccinia coronata pathotypes MQMG and LGCC in the oat (Avena sativa L.) cultivars Ogle and TAM O-301. Theor Appl Genet 116:517–527
Jacobs MM, Vosman B, Vleeshouwers VG, Visser RG, Henken B, van den Berg RG (2010) A novel approach to locate Phytophthora infestans resistance genes on the potato genetic map. Theor Appl Genet 120:785–796
Jellen EN, Gill BS, Cox TS (1994) Genomic differentiates between A/D- and C-genome chromatin and detects intergenomic translocations in polyploidy oat species (genus Avena). Genome 37:613–618
Jin H, Domier LL, Shen X, Kolb FL (2000) Combined AFLP and RFLP mapping in two hexaploid oat recombinant inbred populations. Genome 43:94–101
Jo KR, Arens M, Kim TY, Jongsma MA, Visser RGF, Jacobsen E, Vossen JH (2011) Mapping of the S. demissum late blight resistance gene R8 to a new locus on chromosome IX. Theor Appl Genet 123:1331–1340
Kremer CA, Lee M, Holland JB (2001) A restriction fragment length polymorphism based linkage map of a diploid Avena recombinant inbred line population. Genome 44:192–204
Kuang H, Woo SS, Meyers BC, Nevo E, Michelmore RW (2004) Multiple genetic processes result in heterogeneous rates of evolution within the major cluster disease resistance genes in lettuce. Plant Cell 16:2870–2894
Leal-Bertioli SC, José AC, Alves-Freitas DM, Moretzsohn MC, Guimarães PM, Nielen S, Vidigal BS, Pereira RW, Pike J, Fávero AP, Parniske M, Varshney RK, Bertioli DJ (2009) Identification of candidate genome regions controlling disease resistance in Arachis. BMC Plant Biol 22:9–112
Llorente F, Alonso-Blanco C, Sánchez-Rodriguez C, Jorda L, Molina A (2005) ERECTA receptor-like kinase and heterotrimeric G protein from Arabidopsis are required for resistance to the necrotrophic fungus Plectosphaerella cucumerina. Plant J 43:165–180
Loarce Y, Hueros G, Ferrer E (1996) A molecular linkage map of rye. Theor Appl Genet 93:1112–1118
Loarce Y, Sanz MJ, Irigoyen ML, Fominaya A, Ferrer E (2009) Mapping of STS markers obtained from oat resistance gene analog sequences. Genome 52:608–619
Madsen LH, Collins NC, Rakwalska M, Backes G, Sandal N, Krusell L, Jensen J, Waterman EH, Jahoor A, Ayliffe M, Pryor A, Langridge P, Schulze-Lefert P, Stougaard J (2003) Barley disease resistance gene analogs of the NBS-LRR class: identification and mapping. Mol Genet Genomics 269:150–161
Martin GB, Brommonschenkel SH, Chunwongse J, Frary A, Ganal MW, Spivey R, Wu T, Earle ED, Tanksley SD (1993) Map-based cloning of a protein kinase gene conferring disease resistance in tomato. Science 262:1432–1436
McCartney CA, Stonehouse RG, Rossnagel BG, Eckstein PE, Scoles GJ, Zatorski T, Beattie AD, Chong J (2011) Mapping of the oat crown rust resistance gene Pc91. Theor Appl Genet 122:317–325
McFadden HG, Lehmensiek A, Lagudah ES (2006) Resistance gene analogues of wheat: molecular genetic analysis of ESTs. Theor Appl Genet 113:987–1002
McHale LK, Truco MJ, Kozik A, Wroblewski T, Ochoa OE, Lahre KA, Knapp SJ, Michelmore RW (2009) The genomic architecture of disease resistance in lettuce. Theor Appl Genet 118:565–580
Meyers BC, Chin DB, Shen KA, Sivaramakrishnan S, Lavelle DO, Zhang Z, Michelmore RW (1998) The major resistance gene cluster in lettuce is highly duplicated and spans several megabases. Plant Cell 10:1817–1832
Meyers BC, Kozik A, Griego A, Kuang H, Michelmore RW (2003) Genome-wide analysis of NBS-LRR-encoding genes in Arabidopsis. Plant Cell 15:809–834
O’Donoughue LS, Kianian SF, Rayapati PJ, Penner GA, Sorrells ME, Tanksley SD, Phillips RL, Rines HW, Lee M, Fedak G, Molnar SJ, Hoffman D, Salas CA, Wu B, Autrique E, Van Deynze A (1995) A molecular linkage map of cultivated oat. Genome 38:368–380
Oliver RE, Lazo GR, Lutz JD, Rubenfield MJ, Tinker NA, Anderson JM, Wisniewski Morehead NH, Adhikary D, Jellen EN, Maughan PJ, Brown Guedira GL, Chao S, Beattie AD, Carson ML, Rines HW, Obert DE, Bonman JM, Jackson EW (2011) Model SNP development for complex genomes based on hexaploid oat using high-throughput 454 sequencing technology. BMC Genomics 12:77
Portyanko VA, Hoffman D, Lee M, Holland JB (2001) A linkage map of hexaploid oat based on grass anchor DNA clones and its relationship to other oat maps. Genome 44:249–265
Portyanko VA, Chen G, Rines HW, Phillips RL, Leonard KJ, Ochocki GE, Stuthman DS (2005) Quantitative trait loci for partial resistance to crown rust, Puccinia coronata, in cultivated oat, Avena sativa L. Theor Appl Genet 111:313–324
Rietman H, Bijsterbosch G, Cano L, Lee HR, Vossen J, Jacobsen E, Visser R, Kamoun S, Vleeshouwers V (2012) Qualitative and quantitative late blight resistance in the potato cultivar Sarpo Mira is determined by perception of five distinct RXLR effector. Mol Plant Microbe Interact. doi:10.1094/MPMI-01-12-0010-R)
Rines HW, Porter HL, Carson ML, Ochocki GE (2007) Introgression of crown rust resistance from diploid oat Avena strigosa into hexaploid cultivated oat A. sativa by two methods: direct crosses and through an initial 2x.4x synthetic hexaploid. Euphytica 158:67–79
Romeis T (2001) Protein kinases in the plant defence response. Current Opinion Plant Biol 4:407–414
Sanz MJ, Jellen EN, Loarce Y, Irigoyen ML, Ferrer E, Fominaya A (2010) A new chromosome nomenclature system for oat (Avena sativa L and A. byzantina C. Koch) based on FISH analysis of monosomic lines. Theor Appl Genet 121:1541–1552
Sanz MJ, Loarce Y, Ferrer E, Fominaya A (2012) Use of tyramide-fluorescence in situ hybridization and chromosome microdissection for ascertaining homology relationships and chromosome linkage group associations in oats. Cytogenet Genome Res 136:145–156
Satheeskumar S, Sharp PJ, Lagudah ES, McIntosh RA, Molnar SJ (2011) Genetic association of crown rust resistance gene Pc68, storage protein loci and resistance gene analogs in oats. Genome 54:484–497
Simons MD (1972) Polygenic resistance to plant disease and its use in breeding resistant cultivars. J Environ Qual 1:232–240
Song WY, Wang GL, Chen LL, Kim HS, Pi LY, Holsten T, Gardner J, Wang B, Zhai WX, Zhu LH, Fauquet C, Ronald P (1995) A receptor kinase-like protein encoded by the rice disease resistance gene Xa21. Science 270:1804–1806
Syed NH, Sørensen AP, Antonise R, van de Wiel C, van der Linden CG, van‘t Westende W, Hooftman DAP, den Nijs HCM, Flavell AJ (2006) A detailed linkage map of lettuce based on SSAP, AFLP and NBS markers. Theor Appl Genet 112:517–527
Tameling VIL, Takken FLW (2008) Resistance proteins: scouts of the plant innate immune system. Eur J Plant Pathol 121:243–255
Tinker NA, Kilian A, Wight CP, Heller-Uszynska K, Wenzl P, Rines HW, Bjørnstad A, Howarth CJ, Jannink JL, Anderson JM, Rossnagel BG, Stuthman DD, Sorrells ME, Jackson EW, Tuvesson S, Kolb FL, Olsson O, Federizzi LC, Carson ML, Ohm HW, Molnar SJ, Scoles GJ, Eckstein PE, Bonman JM, Ceplitis A, Langdon T (2008) New DArT markers for oat provide enhanced map coverage and global germplasm characterization. BMC Genomics 21:10–39
van der Linden CG, Wouters D, Mihalka V, Kochieva E, Smulders M, Vosman B (2004) Efficient targeting of plant disease resistance loci using NBS profiling. Theor Appl Genet 109:384–393
van der Linden, Smulders MJM, Vosman BJ (2005) Motif-directed Profiling: a glance at molecular evolution. In: Bakker Chatrou F, Gravendeel LWB, Pelser PB (eds) Plant species-level systematic: new perspectives on patterns and process. Regnum Vegetabile, vol. 143. ARG Gantner Verlag, Rugell, Liechtenstein, Koeltz, Koeningstein, Germany, pp 201–303
Vision TJ, Brown DG, Shmoys DB, Durrett RT, Tanksley SD (2000) Selective mapping: a strategy for optimizing the construction of high-density linkage maps. Genetics 155:407–420
Wang GX, Chen Y, Zhao JR, Li L, Korban SS, Wang FG, Li JS, Dai JRXuML (2007) Mapping of defense response gene homologs and their association with resistance loci in maize. J Integr Plant Biol 49:1580–1598
Whitaker VM, James M, Bradeen JM, Debener T, Biber A, Hokanson SC (2010) Rdr3, a novel locus conferring black spot disease resistance in tetraploid rose: genetic analysis, LRR profiling, and SCAR marker development. Theor Appl Genet 120:573–585
Wight CP, Tinker NA, Kianian SF, Sorrells ME, O’Donoughue LS, Hoffman DL, Groh S, Scoles GL, Li CD, Webster FH, Phillips RL, Rines HW, Livingston SM, Armstrong KC, Fedak G, Molnar SJ (2003) A molecular marker map in ‘Kanota’ × ‘Ogle’ hexaploid oat (Avena sp) enhanced by additional markers and a robust framework. Genome 46:28–47
Yu J, Herrmann M (2006) Inheritance and mapping of a powdery mildew resistance gene introgressed from Avena macrostachya in cultivated oat. Theor Appl Genet 113:429–437
Yu GX, Wise RP (2000) An anchored AFLP- and retrotransposon-based map of diploid Avena. Genome 43:736–749
Yu GX, Braun E, Wise RP (2001) Rds and Rih mediate hypersensitive cell death independent of gene-for-gene resistance to the oat crown rust pathogen Puccinia coronata f. sp. avenae. Mol Plant Microbe Interact 14:1376–1383
Zhang H, Guan H, Li J, Zhu J, Xie C, Zhou Y, Duan X, Yang T, Sun Q, Liu Z (2010) Genetic and comparative genomics mapping reveals that a powdery mildew resistance gene Ml3D232 originating from wild emmer co-segregates with an NBS-LRR analog in common wheat (Triticum aestivum L.). Theor Appl Genet 121:1613–1621
Zhu S, Kaeppler HF (2003) Identification of quantitative trait loci for resistance to crown rust in oat line MAM17-5. Crop Sci 43:358–366
Zhu S, Kolb FL, Kaeppler HF (2003) Molecular mapping of genomic regions underlying barley yellow dwarf tolerance in cultivated oat (Avena sativa L.) Theor Appl Genet 106:1300–1306
Acknowledgments
The authors thank Drs. R. P. Wise, H. W. Rines and N. A. Tinker for providing the RILs and the segregation data for Asw, MN and KO, respectively. Dr. L. H. Madsen is thanked for her gift of barley cv Renata. Dr. Sara Dezhsetan is acknowledged for her kind help with the initial profiling experiments. This work was supported by the Spanish Ministerio de Ciencia e Innovación via grants AGL2006-04165 and AGL2010-17042.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by T. Miedaner.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Sanz, M.J., Loarce, Y., Fominaya, A. et al. Identification of RFLP and NBS/PK profiling markers for disease resistance loci in genetic maps of oats. Theor Appl Genet 126, 203–218 (2013). https://doi.org/10.1007/s00122-012-1974-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00122-012-1974-8


