Effects of Rht-B1 and Ppd-D1 loci on pollinator traits in wheat

  • Takashi OkadaEmail author
  • J. E. A. Ridma M. Jayasinghe
  • Paul Eckermann
  • Nathan S. Watson-Haigh
  • Patricia Warner
  • Yonina Hendrikse
  • Mathieu Baes
  • Elise J. Tucker
  • Hamid Laga
  • Kenji Kato
  • Marc Albertsen
  • Petra Wolters
  • Delphine Fleury
  • Ute Baumann
  • Ryan Whitford
Original Article


Key message

Elite wheat pollinators are critical for successful hybrid breeding. We identified Rht-B1 and Ppd-D1 loci affecting multiple pollinator traits and therefore represent major targets for improving hybrid seed production.


Hybrid breeding has a great potential to significantly boost wheat yields. Ideal male pollinators would be taller in stature, contain many spikelets well-spaced along the spike and exhibit high extrusion of large anthers. Most importantly, flowering time would match with that of the female parent. Available genetic resources for developing an elite wheat pollinator are limited, and the genetic basis for many of these traits is largely unknown. Here, we report on the genetic analysis of pollinator traits using biparental mapping populations. We identified two anther extrusion QTLs of medium effect, one on chromosome 1BL and the other on 4BS coinciding with the semi-dwarfing Rht-B1 locus. The effect of Rht-B1 alleles on anther extrusion is genotype dependent, while tall plant Rht-B1a allele is consistently associated with large anthers. Multiple QTLs were identified at the Ppd-D1 locus for anther length, spikelet number and spike length, with the photoperiod-sensitive Ppd-D1b allele associated with favourable pollinator traits in the populations studied. We also demonstrated that homeoloci, Rht-D1 and Ppd-B1, influence anther length among other traits. These results suggest that combinations of Rht-B1 and Ppd-D1 alleles control multiple pollinator traits and should be major targets of hybrid wheat breeding programs.



This research was supported by DuPont—Pioneer Hi-Bred International. We thank David Correia, Yuriy Onyskiv, Vy Nguyen, Alex Kovalchuk and Dr Ursula Langridge for assisting with glasshouse work. We also thank Drs Radoslaw Suchecki and Beata Sznajder for data analysis and Dr. Ajay Sandhu for critical advice for the project. We also thank Margaret Pallotta for technical advice and critical reading and editing of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standards

The authors declare that this study complies with the current laws of the countries in which the experiments were performed.

Supplementary material

122_2019_3329_MOESM1_ESM.pdf (545 kb)
Effect of severe dwarf (SD) on pollinator traits and data analysis including SD plants (PDF 546 kb)
122_2019_3329_MOESM2_ESM.pdf (1.2 mb)
Supplemental Figures S1–S9 (PDF 1251 kb)
122_2019_3329_MOESM3_ESM.xlsx (483 kb)
Table S1. KASPTM markers used for developing genetic linkage map for populations #1 and #2. Table S2. GBS markers used for developing genetic linkage map for population #1. Table S3. GBS markers used for developing genetic linkage map for population #2. Table S4. Summary information for genetic linkage map and QTL analysis for population #1 and #2. Table S5. A list of genetic loci associated with anther extrusion reported in the previous publications. Table S6. Phenology genes mapped on Chinese Spring reference sequence IWGSC RefSeq v1.0 to compare physical location of genetic loci associated with pollinator traits. Table S7. Summary statistics for evaluated traits in Rht-1 and Ppd-1 NILs used in this study. Table S8. Physical location of AE loci and flowering time (FT) or plant/floral architecture (FA) genes/loci in CS reference map IWGSC RefSeq v1.0 (XLSX 483 kb)


  1. Abe M, Kobayashi Y, Yamamoto S, Daimon Y, Yamaguchi A, Ikeda Y, Ichinoki H, Notaguchi M, Goto K, Araki T (2005) FD, a bZIP protein mediating signals from the floral pathway integrator FT at the shoot apex. Science 309:1052–1056CrossRefGoogle Scholar
  2. Alghabari F, Lukac M, Jones HE, Gooding MJ (2014) Effect of Rht alleles on the tolerance of wheat grain set to high temperature and drought stress during booting and anthesis. J Agron Crop Sci 200:36–45CrossRefGoogle Scholar
  3. Beales J, Turner A, GriYths S, Snape JW, Laurie DA (2007) A Pseudo-Response Regulator is misexpressed in the photoperiod insensitive Ppd-D1a mutant of wheat (Triticum aestivum L.). Theor Appl Genet 115:721–733CrossRefGoogle Scholar
  4. Binghua L, Jingyang D (1986) A dominant gene for male-sterility in wheat. Plant Breed 97:204–209CrossRefGoogle Scholar
  5. Boden SA, Cavanagh C, Cullis BR, Ramm K, Greenwood J, Jean Finnegan E, Trevaskis B, Swain SM (2015) Ppd-1 is a key regulator of inflorescence architecture and paired spikelet development in wheat. Nat Plants 1:14016CrossRefGoogle Scholar
  6. Boeven PHG, Longin CFH, Leiser WL, Kollers S, Ebmeyer E, Wurschum T (2016) Genetic architecture of male floral traits required for hybrid wheat breeding. Theor Appl Genet 129:2343–2357CrossRefGoogle Scholar
  7. Boeven PHG, Würschum T, Rudloff J, Ebmeyer E, Longin CFH (2018) Hybrid seed set in wheat is a complex trait but can be improved indirectly by selection for male floral traits. Euphytica 214:110CrossRefGoogle Scholar
  8. Broman KW, Wu H, Sen S, Churchill GA (2003) R/qtl: QTL mapping in experimental crosses. Bioinformatics 19:889–890CrossRefGoogle Scholar
  9. Buerstmayr M, Buerstmayr H (2015) Comparative mapping of quantitative trait loci for Fusarium head blight resistance and anther retention in the winter wheat population Capo × Arina. Theor Appl Genet 128:1519–1530CrossRefGoogle Scholar
  10. Buerstmayr M, Buerstmayr H (2016) The semidwarfing alleles Rht-D1b and Rht-B1b show marked differences in their associations with anther-retention in wheat heads and with Fusarium head blight susceptibility. Phytopathology 106:1544–1552CrossRefGoogle Scholar
  11. Cheng Y, Dai X, Zhao Y (2006) Auxin biosynthesis by the YUCCA flavin monooxygenases controls the formation of floral organs and vascular tissues in Arabidopsis. Genes Dev 20:1790–1799CrossRefGoogle Scholar
  12. Daviere JM, Achard P (2016) A pivotal role of DELLAs in regulating multiple hormone signals. Mol Plant 9:10–20CrossRefGoogle Scholar
  13. Distelfeld A, Li C, Dubcovsky J (2009) Regulation of flowering in temperate cereals. Curr Opin Plant Biol 12:178–184CrossRefGoogle Scholar
  14. Elshire RJ, Glaubitz JC, Sun Q, Poland JA, Kawamoto K, Buckler ES, Mitchell SE (2011) A robust, simple genotyping-by-sequencing (GBS) approach for high diversity species. PLoS ONE 6:e19379CrossRefGoogle Scholar
  15. FAO (2015) Statistical pocketbook world food and agriculture 2015. Food and Agriculture Organization of the United Nations. Accessed 25 Nov 2018
  16. Ghanem ME, Marrou H, Sinclair TR (2015) Physiological phenotyping of plants for crop improvement. Trends Plant Sci 20:139–144CrossRefGoogle Scholar
  17. Gilmour A, Gogel B, Cullis B, Thompson R (2009) ASReml user guide release 3.0. VSN International Ltd. Accessed 25 Nov 2018
  18. Gils M, Kempe K, Boudichevskaia A, Jerchel R, Pescianschi D, Schmidt R, Kirchhoff M, Schachschneider R (2013) Quantitative assessment of wheat pollen shed by digital image analysis of trapped airborne pollen grains. Adv Crop Sci Technol 2:119Google Scholar
  19. Griffiths S, Dunford RP, Coupland G, Laurie DA (2003) The evolution of CONSTANS-like gene families in barley, rice, and Arabidopsis. Plant Physiol 131:1855–1867CrossRefGoogle Scholar
  20. Guo Z, Song Y, Zhou R, Ren Z, Jia J (2010) Discovery, evaluation and distribution of haplotypes of the wheat Ppd-D1 gene. New Phytol 185:841–851CrossRefGoogle Scholar
  21. Halliwell J, Borrill P, Gordon A, Kowalczyk R, Pagano ML, Saccomanno B, Bentley AR, Uauy C, Cockram J (2016) Systematic investigation of FLOWERING LOCUS T-like Poaceae gene families identifies the short-day expressed flowering pathway gene, TaFT3 in wheat (Triticum aestivum L.). Plant Sci 7:857Google Scholar
  22. He X, Lillemo M, Shi JR, Wu JR, Bjornstad A, Belova T, Dreisigacker S, Duveiller E, Singh P (2016a) QTL characterization of Fusarium head blight resistance in CIMMYT bread wheat line Soru#1. PLoS ONE 11:e0158052CrossRefGoogle Scholar
  23. He X, Singh PK, Dreisigacker S, Singh S, Lillemo M, Duveiller E (2016b) Dwarfing genes Rht-B1b and Rht-D1b are associated with both Type I FHB susceptibility and low anther extrusion in two bread wheat populations. PLoS ONE 11:e0162499CrossRefGoogle Scholar
  24. International Wheat Genome Sequencing C (2018) Shifting the limits in wheat research and breeding using a fully annotated reference genome. Science 361:eaar7191CrossRefGoogle Scholar
  25. Jimenez-Berni JA, Deery DM, Rozas-Larraondo P, Condon ATG, Rebetzke GJ, James RA, Bovill WD, Furbank RT, Sirault XRR (2018) High throughput determination of plant height, ground cover, and above-ground biomass in wheat with LiDAR. Front Plant Sci 9:237CrossRefGoogle Scholar
  26. Kneipp J (2017) Control of Fusarium head blight in northern NSW. Accessed 25 Nov 2018
  27. Kosuge K, Watanabe N, Kuboyama T, Melnik VM, Yanchenko VI, Rosova MA, Goncharov NP (2008) Cytological and microsatellite mapping of mutant genes for spherical grain and compact spikes in durum wheat. Euphytica 159:289–296CrossRefGoogle Scholar
  28. Kowalski AM, Gooding M, Ferrante A, Slafer GA, Orford S, Gasperini D, Griffiths S (2016) Agronomic assessment of the wheat semi-dwarfing gene Rht8 in contrasting nitrogen treatments and water regimes. Field Crops Res 191:150–160CrossRefGoogle Scholar
  29. Langer SM, Longin CFH, Wurschum T (2014) Phenotypic evaluation of floral and flowering traits with relevance for hybrid breeding in wheat (Triticum aestivum L.). Plant Breed 133:433–441CrossRefGoogle Scholar
  30. Li C, Dubcovsky J (2008) Wheat FT protein regulates VRN1 transcription through interactions with FDL2. Plant J 55:543–554CrossRefGoogle Scholar
  31. Longin CFH, Muhleisen J, Maurer HP, Zhang HL, Gowda M, Reif JC (2012) Hybrid breeding in autogamous cereals. Theor Appl Genet 125:1087–1096CrossRefGoogle Scholar
  32. Longin CF, Gowda M, Muhleisen J, Ebmeyer E, Kazman E, Schachschneider R, Schacht J, Kirchhoff M, Zhao Y, Reif JC (2013) Hybrid wheat: quantitative genetic parameters and consequences for the design of breeding programs. Theor Appl Genet 126:2791–2801CrossRefGoogle Scholar
  33. Lu Q, Lillemo M, Skinnes H, He X, Shi J, Ji F, Dong Y, Bjornstad A (2013) Anther extrusion and plant height are associated with Type I resistance to Fusarium head blight in bread wheat line ‘Shanghai-3/Catbird’. Theor Appl Genet 126:317–334CrossRefGoogle Scholar
  34. Manske GGB, Ortiz-Monasterio JI, van Ginkel RM, Rajaram S, Vlek PLG (2002) Phosphorus use efficiency in tall, semi-dwarf and dwarf near-isogenic lines of spring wheat. Euphytica 125:113–119CrossRefGoogle Scholar
  35. Miedaner T, Schulthess AW, Gowda M, Reif JC, Longin CF (2017) High accuracy of predicting hybrid performance of Fusarium head blight resistance by mid-parent values in wheat. Theor Appl Genet 130:461–470CrossRefGoogle Scholar
  36. Milohnic J, Jost M (1970) Pollen production and anther extrusion of wheat (Triticum aestivum L. Em Thell.). Acta Agron Acad Sci Hung 19:17–23Google Scholar
  37. Mulki MA, Bi X, von Korff M (2018) FLOWERING LOCUS T3 controls spikelet initiation but not floral development. Plant Physiol 178:1170–1186CrossRefGoogle Scholar
  38. Muqaddasi QH, Lohwasser U, Nagel M, Borner A, Pillen K, Roder MS (2016) Genome-wide association mapping of anther extrusion in hexaploid spring wheat. PLoS ONE 11:e0155494CrossRefGoogle Scholar
  39. Muqaddasi QH, Brassac J, Borner A, Pillen K, Roder MS (2017a) Genetic architecture of anther extrusion in spring and winter wheat. Front Plant Sci 8:754CrossRefGoogle Scholar
  40. Muqaddasi QH, Pillen K, Plieske J, Ganal MW, Roder MS (2017b) Genetic and physical mapping of anther extrusion in elite European winter wheat. PLoS ONE 12:e0187744CrossRefGoogle Scholar
  41. Nemoto Y, Kisaka M, Fuse T, Yano M, Ogihara Y (2003) Characterization and functional analysis of three wheat genes with homology to the CONSTANS flowering time gene in transgenic rice. Plant J 36:82–93CrossRefGoogle Scholar
  42. Nguyen V, Fleury D, Timmins A, Laga H, Hayden M, Mather D, Okada T (2015) Addition of rye chromosome 4R to wheat increases anther length and pollen grain number. Theor Appl Genet 128:953–964CrossRefGoogle Scholar
  43. Okada T, Whitford R (2019) Hybrid wheat and abiotic stress. In: Rajpal VR, Sehgal D, Kumar A, Raina SN (eds) Genomics assisted breeding of crops for abiotic stress tolerance, vol 2. Sustainable development and biodiversity 21. Springer, Switzerland.
  44. Okada T, Jayasinghe J, Nansamba M, Baes M, Warner P, Kouidri A, Correia D, Nguyen V, Whitford R, Baumann U (2018) Unfertilized ovary pushes wheat flower open for cross-pollination. J Exp Bot 69:399–412CrossRefGoogle Scholar
  45. Pearce S, Saville R, Vaughan SP, Chandler PM, Wilhelm EP, Sparks CA, Al-Kaff N, Korolev A, Boulton MI, Phillips AL, Hedden P, Nicholson P, Thomas SG (2011) Molecular characterization of Rht-1 dwarfing genes in hexaploid wheat. Plant Physiol 157:1820–1831CrossRefGoogle Scholar
  46. Peng J, Richards DE, Hartley NM, Murphy GP, Devos KM, Flintham JE, Beales J, Fish LJ, Worland AJ, Pelica F, Sudhakar D, Christou P, Snape JW, Gale MD, Harberd NP (1999) ‘Green revolution’ genes encode mutant gibberellin response modulators. Nature 400:256–261CrossRefGoogle Scholar
  47. Pickett A (1993) Hybrid wheat results and problems. Paul Parey Scientific, BerlinGoogle Scholar
  48. Poland J, Endelman J, Dawson J, Rutkoski J, Wu SY, Manes Y, Dreisigacker S, Crossa J, Sanchez-Villeda H, Sorrells M, Jannink JL (2012) Genomic selection in wheat breeding using genotyping-by-sequencing. Plant Genome 5:103–113CrossRefGoogle Scholar
  49. Rebetzke GJ, Richards RA, Fettell NA, Long M, Condon AG, Forrester RI, Botwright TL (2007) Genotypic increases in coleoptile length improves stand establishment, vigour and grain yield of deep-sown wheat. Field Crops Res 100:10–23CrossRefGoogle Scholar
  50. Richards RA (1992) The effect of dwarfing genes in spring wheat in dry environments. 1. Agronomic characteristics. Aust J Agric Res 43:517–527CrossRefGoogle Scholar
  51. Rogowsky PM, Sorrels ME, Shepherd KW, Langridge P (1993) Characterization of wheat-rye recombinants with RFLP and PCR probes. Theor Appl Genet 85:1023–1028CrossRefGoogle Scholar
  52. RStudio_Team (2015) RStudio: integrated development for R. RStudio, Inc. Accessed 25 Nov 2018
  53. Sasakuma T, Maan SS, Williams ND (1978) EMS-induced male-sterile mutants in euplasmic and alloplasmic common wheat. Crop Sci 18:850–853CrossRefGoogle Scholar
  54. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) FIJI: an open-source platform for biological-image analysis. Nat Methods 9:676–682CrossRefGoogle Scholar
  55. Shaw LM, Turner AS, Laurie DA (2012) The impact of photoperiod insensitive Ppd-1a mutations on the photoperiod pathway across the three genomes of hexaploid wheat (Triticum aestivum). Plant J 71:71–84CrossRefGoogle Scholar
  56. Skinnes H, Semagn K, Tarkegne Y, Maroy AG, Bjornstad A (2010) The inheritance of anther extrusion in hexaploid wheat and its relationship to Fusarium head blight resistance and deoxynivalenol content. Plant Breed 129:149–155CrossRefGoogle Scholar
  57. Song X, Feng J, Cui Z, Zhang C, Sun D (2018) Genome-wide association study for anther length in some elite bread wheat germplasm. Czech J Genet Plant Breed 54:109–114CrossRefGoogle Scholar
  58. Tanio M, Kato K (2007) Development of near-isogenic lines for photoperiod-insensitive genes, Ppd-B1 and Ppd-D1, carried by the Japanese wheat cultivars and their effect on apical development. Breed Sci 57:65–72CrossRefGoogle Scholar
  59. Taylor J, Butler D (2017) R package ASMap: efficient genetic linkage map construction and diagnosis. J Stat Softw 79:1–29CrossRefGoogle Scholar
  60. Tester M, Langridge P (2010) Breeding technologies to increase crop production in a changing world. Science 327:818–822CrossRefGoogle Scholar
  61. Tricker PJ, ElHabti A, Schmidt J, Fleury D (2018) The physiological and genetic basis of combined drought and heat tolerance in wheat. J Exp Bot 69:3195–3210CrossRefGoogle Scholar
  62. Turner A, Beales J, Faure S, Dunford RP, Laurie DA (2005) The pseudo-response regulator Ppd-H1 provides adaptation to photoperiod in barley. Science 310:1031–1034CrossRefGoogle Scholar
  63. Voorrips RE (2002) MapChart: software for the graphical presentation of linkage maps and QTLs. J Hered 93:77–78CrossRefGoogle Scholar
  64. VSN_International (2011) GenStat for windows, 14th edn. VSN International, Hemel HempsteadGoogle Scholar
  65. Whitford R, Fleury D, Reif JC, Garcia M, Okada T, Korzun V, Langridge P (2013) Hybrid breeding in wheat: technologies to improve hybrid wheat seed production. J Exp Bot 64:5411–5428CrossRefGoogle Scholar
  66. Wilhelm EP, Boulton MI, Al-Kaff N, Balfourier F, Bordes J, Greenland AJ, Powell W, Mackay IJ (2013) Rht-1 and Ppd-D1 associations with height, GA sensitivity, and days to heading in a worldwide bread wheat collection. Theor Appl Genet 126:2233–2243CrossRefGoogle Scholar
  67. Wilkinson PA, Winfield MO, Barker GLA, Allen AM, Burridge A, Coghill JA, Edwards KJ (2012) CerealsDB 2.0: an integrated resource for plant breeders and scientists. BMC Bioinformatics 13:219CrossRefGoogle Scholar
  68. Würschum T, Liu G, Boeven PHG, Longin CFH, Mirdita V, Kazman E, Zhao Y, Reif JC (2018) Exploiting the Rht portfolio for hybrid wheat breeding. Theor Appl Genet 131:1433–1442CrossRefGoogle Scholar
  69. Yan L, Loukoianov A, Tranquilli G, Helguera M, Fahima T, Dubcovsky J (2003) Positional cloning of the wheat vernalization gene VRN1. Proc Natl Acad Sci USA 100:6263–6268CrossRefGoogle Scholar
  70. Yan L, Loukoianov A, Blechl A, Tranquilli G, Ramakrishna W, SanMiguel P, Bennetzen JL, Echenique V, Dubcovsky J (2004) The wheat VRN2 gene is a flowering repressor down-regulated by vernalization. Science 303:1640–1644CrossRefGoogle Scholar
  71. Yan L, Fu D, Li C, Blechl A, Tranquilli G, Bonafede M, Sanchez A, Valarik M, Yasuda S, Dubcovsky J (2006) The wheat and barley vernalization gene VRN3 is an orthologue of FT. Proc Natl Acad Sci USA 103:19581–19586CrossRefGoogle Scholar
  72. Youssefian S, Kirby EJM, Gale MD (1992) Pleiotropic effects of the Ga-Insensitive Rht dwarfing genes in wheat. 2. Effects on leaf, stem, ear and floret growth. Field Crops Res 28:179–190CrossRefGoogle Scholar
  73. Zadoks JC, Chang TT, Konzak CF (1974) Decimal code for growth stages of cereals. Weed Res 14:415–421CrossRefGoogle Scholar
  74. Zhang XK, Xiao YG, Zhang Y, Xia XC, Dubcovsky J, He ZH (2008) Allelic variation at the vernalization genes Vrn-A1, Vrn-B1, Vrn-D1, and Vrn-B3 in Chinese wheat cultivars and their association with growth habit. Crop Sci 48:458–470CrossRefGoogle Scholar
  75. Zhao Y, Li Z, Liu G, Jiang Y, Maurer HP, Wurschum T, Mock HP, Matros A, Ebmeyer E, Schachschneider R, Kazman E, Schacht J, Gowda M, Longin CF, Reif JC (2015) Genome-based establishment of a high-yielding heterotic pattern for hybrid wheat breeding. Proc Natl Acad Sci USA 112:15624–15629Google Scholar
  76. Zhao XY, Hong P, Wu JY, Chen XB, Ye XG, Pan YY, Wang J, Zhang XS (2016) The tae-miR408-mediated control of TaTOC1 genes transcription is required for the regulation of heading time in wheat. Plant Physiol 170:1578–1594Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Agriculture, Food and Wine, Plant Genomics CentreUniversity of AdelaideUrrbraeAustralia
  2. 2.College of Science, Health, Engineering and EducationMurdoch UniversityMurdochAustralia
  3. 3.Phenomics and Bioinformatics Research CentreUniversity of South AustraliaMawson LakesAustralia
  4. 4.Graduate School of Environmental and Life ScienceOkayama UniversityOkayamaJapan
  5. 5.DuPont-Pioneer Hi-Bred International Inc.JohnstonUSA

Personalised recommendations