Plant Molecular Biology

, Volume 88, Issue 4–5, pp 487–502 | Cite as

Comparison of gene expression profiles and responses to zinc chloride among inter- and intraspecific hybrids with growth abnormalities in wheat and its relatives

  • Kiyofumi Takamatsu
  • Julio C. M. Iehisa
  • Ryo Nishijima
  • Shigeo TakumiEmail author


Hybrid necrosis is a well-known reproductive isolation mechanism in plant species, and an autoimmune response is generally considered to trigger hybrid necrosis through epistatic interaction between disease resistance-related genes in hybrids. In common wheat, the complementary Ne1 and Ne2 genes control hybrid necrosis, defined as type I necrosis. Two other types of hybrid necrosis (type II and type III) have been observed in interspecific hybrids between tetraploid wheat and Aegilops tauschii. Another type of hybrid necrosis, defined here as type IV necrosis, has been reported in F1 hybrids between Triticum urartu and some accessions of Triticum monococcum ssp. aegilopoides. In types I, III and IV, cell death occurs gradually starting in older tissues, whereas type II necrosis symptoms occur only under low temperature. To compare comprehensive gene expression patterns of hybrids showing growth abnormalities, transcriptome analysis of type I and type IV necrosis was performed using a wheat 38k oligo-DNA microarray. Defense-related genes including many WRKY transcription factor genes were dramatically up-regulated in plants showing type I and type IV necrosis, similarly to other known hybrid abnormalities, suggesting an association with an autoimmune response. Reactive oxygen species generation and necrotic cell death were effectively inhibited by ZnCl2 treatment in types I, III and IV necrosis, suggesting a significant association of Ca2+ influx in upstream signaling of necrotic cell death in wheat hybrid necrosis.


Calcium influx Microarray Postzygotic barrier Programmed cell death Reactive oxygen species Wheat 



We thank emeritus professor Dr. Koichiro Tsunewaki for supplying seeds of NILs, Ne1-S615 and Ne2-S615. We are grateful to Professor Dr. Hitoshi Nakayashiki for his useful suggestions. The diploid wheat seeds used in this study were supplied by the National BioResource Project-Wheat, Japan ( This work was supported by Grants-in-Aid for Scientific Research (B) Nos. 21380005 and 25292008 from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Supplementary material

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Supplementary material 1 (PDF 100 kb)
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Supplementary material 2 (XLSX 848 kb)
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  1. Alcázar R, García AV, Parker JE, Reymond M (2009) Incremental steps toward incompatibility revealed by Arabidopsis epistatic interactions modulating salicylic acid pathway activation. Proc Natl Acad Sci USA 106:334–339PubMedCentralPubMedCrossRefGoogle Scholar
  2. Alcázar R, García AV, Kronholm I, de Meaux J, Koornneef M, Parker JE, Reymond M (2010) Natural variation at Strubbelig Receptor Kinase 3 drives immune-triggered incompatibilities between Arabidopsis thaliana accessions. Nat Genet 42:1135–1139PubMedCrossRefGoogle Scholar
  3. Bindschedler LV, Minibayeva F, Gardner SL, Gerrish C, Davies DR, Bolwell GP (2001) Early signalling events in the apoplastic oxidative burst in suspension cultured French bean cells involve camp and Ca2+. New Phytol 151:185–194CrossRefGoogle Scholar
  4. Bomblies K, Weigel D (2007) Hybrid necrosis: autoimmunity as a potential gene-flow barrier in plant species. Nat Rev Genet 8:382–393PubMedCrossRefGoogle Scholar
  5. Bomblies K, Lempe J, Epple P, Warthmann N, Lanz C, Dangl JL, Weigel D (2007) Autoimmune response as a mechanism for Dobzhansky–Muller-type incompatibility syndrome in plants. PLoS Biol 5:e236PubMedCentralPubMedCrossRefGoogle Scholar
  6. Brandolini A, Vaccino P, Boggini G, Özkan H, Kilian B, Salamini F (2006) Quantification of genetic relationships among A genomes of wheats. Genome 49:297–305PubMedCrossRefGoogle Scholar
  7. Castagna R, Maga G, Perenzin M, Heun M, Salamini F (1994) RFLP-based genetic relationships of einkorn wheats. Theor Appl Genet 88:818–823PubMedCrossRefGoogle Scholar
  8. Castagna R, Gnocchi S, Perenzin M, Heun M (1997) Genetic variability of the wild diploid wheat Triticum urartu revealed by RFLP and RAPD markers. Theor Appl Genet 94:424–430CrossRefGoogle Scholar
  9. Chae E, Bomblies K, Kim ST, Karelina D, Zaidem M, Ossowski S, Martín-Pizarro C, Laitinen RAE, Rowan BA, Tenenboim H, Lechner S, Demar M, Habring-Müller A, Lanz C, Rätsch G, Weigel D (2014) Species-wide genetic incompatibility analysis identifies immune genes as hot spots of deleterious epistasis. Cell 159:1341–1351PubMedCrossRefGoogle Scholar
  10. Chu CG, Faris JD, Friesen TL, Xu SS (2006) Molecular mapping of hybrid necrosis genes Ne1 and Ne2 in hexaploid wheat using microsatellite markers. Theor Appl Genet 112:1374–1381PubMedCrossRefGoogle Scholar
  11. Conesa A, Götz S, García-Gómez JM, Terol J, Talón M, Robles M (2005) Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics 21:3674–3676PubMedCrossRefGoogle Scholar
  12. Dalal M, Khanna-Chopra R (2001) Differential response of antioxidant enzymes in leaves of necrotic wheat hybrids and their parents. Physiol Plant 111:297–304PubMedCrossRefGoogle Scholar
  13. Davies DR, Bindschedler LV, Strickland TS, Bolwell GP (2006) Production of reactive oxygen species in Arabidopsis thaliana cell suspension cultures in response to an elicitor from Fusarium oxysporum: implications for basal resistance. J Exp Bot 57:1817–1827PubMedCrossRefGoogle Scholar
  14. Dhaliwal HS (1977) Basis of difference between reciprocal crosses involving Triticum boeoticum and T. urartu. Theor Appl Genet 49:283–286PubMedCrossRefGoogle Scholar
  15. Fricano A, Brandolini A, Rossini L, Sourdille P, Wunder J, Effgene S, Hidalgo A, Erba D, Piffanelli P, Salamini F (2014) Crossability of Triticum urartu and Triticum monococcum wheats, homoeologous recombination and description of a panel of interspecific introgression lines. G3 4:1931–1941PubMedCentralPubMedCrossRefGoogle Scholar
  16. Gill BS, Waines JG (1978) Paternal regulation of seed development in wheat hybrids. Theor Appl Genet 51:265–270PubMedCrossRefGoogle Scholar
  17. Hatano H, Mizuno N, Matsuda R, Shitsukawa N, Park P, Takumi S (2012) Dysfunction of mitotic cell division at shoot apices triggered severe growth abortion in interspecific hybrids between tetraploid wheat and Aegilops tauschii. New Phytol 194:1143–1154PubMedCrossRefGoogle Scholar
  18. Hermsen JGT (1963) The genetic basis of hybrid necrosis in wheat. Genetica 33:245–287CrossRefGoogle Scholar
  19. Hoat TX, Nakayashiki H, Tosa Y, Mayama S (2006) Specific cleavage of ribosomal RNA and mRNA during victorin-induced apoptotic cell death in oat. Plant J 46:922–933PubMedCrossRefGoogle Scholar
  20. Ishihama N, Yoshioka H (2012) Post-translational regulation of WRKY transcription factors in plant immunity. Curr Opin Plant Biol 15:431–437PubMedCrossRefGoogle Scholar
  21. Jeuken MJW, Zhang NW, McHale LK, Pelgrom K, den Boer E, Lindhout P, Michelmore RW, Visser RGF, Niks RE (2009) Rin4 causes hybrid necrosis and race-specific resistance in an interspecific lettuce hybrid. Plant Cell 21:3368–3378PubMedCentralPubMedCrossRefGoogle Scholar
  22. Johnson BL, Dhaliwal HS (1976) Reproductive isolation of Triticum boeoticum and Triticum urartu and the origin of the tetraploid wheat. Am J Bot 63:1088–1094CrossRefGoogle Scholar
  23. Kawaura K, Mochida K, Ogihara Y (2008) Genome-wide analysis for identification of salt-responsive genes in common wheat. Funct Integr Genomics 8:277–286PubMedCrossRefGoogle Scholar
  24. Khanna-Chopra R, Dalal M, Kumar PG, Laloraya M (1998) A genetic system involving superoxide causes F1 necrosis in wheat (Triticum aestivum L.). Biochem Biophys Res Commun 248:712–715PubMedCrossRefGoogle Scholar
  25. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  26. Lucas A (2011) amap: another multidimentional analysis package. R package version 0.8-7.
  27. Ma W, Berkowitz GA (2007) The grateful dead: calcium and cell death in plant innate immunity. Cell Microbiol 9:2571–2585PubMedCrossRefGoogle Scholar
  28. Masuda Y, Yamada T, Kuboyama T, Marubashi W (2007) Identification and characterization of genes involved in hybrid lethality in hybrid tobacco cells (Nicotiana suaveolens x N. tabacum) using suppression subtractive hybridization. Plant Cell Rep 26:1595–1604PubMedCrossRefGoogle Scholar
  29. Matsuoka Y (2011) Evolution of polyploid Triticum wheats under cultivation: the role of domestication, natural hybridization and allopolyploid speciation in their diversification. Plant Cell Physiol 52:750–764PubMedCrossRefGoogle Scholar
  30. Matsuoka Y, Takumi S, Kawahara T (2007) Natural variation for fertile triploid F1 formation in allohexaploid wheat speciation. Theor Appl Genet 115:509–518PubMedCrossRefGoogle Scholar
  31. Mizumoto K, Hirosawa S, Nakamura C, Takumi S (2002) Nuclear and chloroplast genome genetic diversity in the wild einkorn wheat, Triticum urartu, revealed by AFLP and SSLP analyses. Hereditas 137:208–214CrossRefGoogle Scholar
  32. Mizuno N, Hosogi N, Park P, Takumi S (2010) Hypersensitive response-like reaction is associated with hybrid necrosis in interspecific crosses between tetraploid wheat and Aegilops tauschii Coss. PLoS One 5:e11326PubMedCentralPubMedCrossRefGoogle Scholar
  33. Mizuno N, Shitsukawa N, Hosogi N, Park P, Takumi S (2011) Autoimmune response and repression of mitotic cell division occur in inter-specific crosses between tetraploid wheat and Aegilops tauschii Coss. that show low temperature-induced hybrid necrosis. Plant J 68:114–128PubMedCrossRefGoogle Scholar
  34. Mur LAJ, Kenton P, Lioyd AJ, Ougham H, Prats E (2008) The hypersensitive response; the centenary is upon us but how much do we know? J Exp Bot 59:501–520PubMedCrossRefGoogle Scholar
  35. Myhre S, Tveit H, Mollestad T, Laegreid A (2006) Additional gene ontology structure for improved biological reasoning. Bioinformatics 22:2020–2027PubMedCrossRefGoogle Scholar
  36. Nakano H, Mizuno N, Tosa Y, Yoshida K, Park P, Takumi S (2015) Accelerated senescence and enhanced disease resistance in hybrid chlorosis lines derived from interspecific crosses between tetraploid wheat and Aegilops tauschii. PLoS One. doi: 10.1371/journal.pone.0121583 Google Scholar
  37. Nishikawa K (1960) Hybrid lethality in crosses between Emmer wheats and Aegilops squarrosa, I. Vitality of F1 hybrids between emmer wheats and Ae. squarrosa var. typica. Seiken Ziho 11:21–28Google Scholar
  38. Nishikawa K (1962a) Hybrid lethality in crosses between Emmer wheats and Aegilops squarrosa, II. Synthesized 6x wheats employed as test varieties. Jpn J Genet 37:227–236CrossRefGoogle Scholar
  39. Nishikawa K (1962b) Hybrid lethality in crosses between Emmer wheats and Aegilops squarrosa, III. Gene analysis of type-2 necrosis. Seiken Ziho 14:45–50Google Scholar
  40. Orrenius S, Zhivotovsky B, Nicotera P (2003) Regulation of cell death: the calcium-apoptosis link. Nat Rev Mol Cell Biol 4:552–565PubMedCrossRefGoogle Scholar
  41. Presgraves DC (2010) The molecular evolutionary basis of species formation. Nat Rev Genet 11:175–180PubMedCrossRefGoogle Scholar
  42. Pukhalskiy VA, Martynov SP, Dobrotvorskaya TV (2000) Analysis of geographical and breeding-related distribution of hybrid necrosis genes in bread wheat (Triticum aestivum L.). Euphytica 114:233–240CrossRefGoogle Scholar
  43. R Core Team (2012) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria.
  44. Rieseberg LH, Willis JH (2007) Plant speciation. Science 317:910–914PubMedCentralPubMedCrossRefGoogle Scholar
  45. Roy RP (1955) Semi-lethal hybrids in crosses of species and synthetic amphidiploids of Tritium and Aegilops. Indian J Genet Plant Breed 14:88–98Google Scholar
  46. Rushton PJ, Somssich IE, Ringler P, Shen QJ (2010) WRKY transcription factors. Trend Plant Sci 15:247–258CrossRefGoogle Scholar
  47. Sears ER (1944) Inviability of intergeneric hybrids involving Triticum monococcum and T. aegilopoides. Genetics 29:113–127PubMedCentralPubMedGoogle Scholar
  48. Song CJ, Steinebrunner I, Wang X, Stout SC, Roux SJ (2006) Extracellular ATP induces the accumulation of superoxide via NADPH oxidases in Arabidopsis. Plant Physiol 140:1222–1232PubMedCentralPubMedCrossRefGoogle Scholar
  49. Sugie A, Murai K, Takumi S (2007) Alteration of respiration capacity and transcript accumulation levels of alternative oxidase genes in necrosis lines of common wheat. Genes Genet Syst 82:231–239PubMedCrossRefGoogle Scholar
  50. Takumi S, Mizuno N (2011) Low temperature-induced necrosis shows phenotypic plasticity in wheat triploid hybrids. Plant Signal Behav 6:1431–1433PubMedCentralPubMedCrossRefGoogle Scholar
  51. Takumi S, Motomura Y, Iehisa JCM, Kobayashi F (2013) Segregation distortion caused by weak hybrid necrosis in recombinant inbred lines of common wheat. Genetica 141:463–470PubMedCrossRefGoogle Scholar
  52. Tsunewaki K (1960) Monosomic and conventional analyses in common wheat. III. Lethality. Jpn J Genet 35:71–75CrossRefGoogle Scholar
  53. Tsunewaki K (1970) Necrosis and chlorosis genes in common wheat and its ancestral species. Seiken Ziho 22:67–75Google Scholar
  54. Tsunewaki K (1992) Aneuploid analyses of hybrid necrosis and hybrid chlorosis in tetraploid wheats using the D genome chromosome substitution lines of durum wheat. Genome 35:594–601CrossRefGoogle Scholar
  55. Tsunewaki K, Koba T (1979) Production and genetic characterization of the co-isogenic lines of a common wheat Triticum aestivum cv. S-615 for ten major genes. Euphytica 28:579–592CrossRefGoogle Scholar
  56. Warnes GR (2012) gplots: various R programming tools for plotting data. R package version 2.11.0.
  57. Wojtaszek P (1997) Oxidative burst: an early plant response to pathogen infection. Biochem J 322:681–692PubMedCentralPubMedGoogle Scholar
  58. Yamagishi Y (1987) Phylogenetic differentiation between two species of the wild diploid wheats. Genbunsha, Kyoto, Japan. ISBN4-87609-144-7Google Scholar
  59. Yamamoto E, Takashi T, Morinaka Y, Lin S, Wu J, Matsumoto T, Kitano H, Matsuoka M, Ashikari M (2010) Gain of deleterious function caused an autoimmune response and Bateson–Dobzhansky–Muller incompatibility in rice. Mol Genet Genomics 283:305–315PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Kiyofumi Takamatsu
    • 1
  • Julio C. M. Iehisa
    • 1
  • Ryo Nishijima
    • 1
  • Shigeo Takumi
    • 1
    Email author
  1. 1.Laboratory of Plant Genetics, Graduate School of Agricultural ScienceKobe UniversityKobeJapan

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