Skip to main content
SpringerLink
Log in

In vitro Culture for Doubled Haploids: Tools for Molecular Breeding

  • Protocol
  • First Online:
Crop Breeding

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1145))

Abstract

Doubled haploid (DH) techniques using immature microspores or embryos rescued in vitro following interspecific hybridization are important tools for breeding wheat and barley and for generating populations for gene mapping purposes. Here, we describe methods being used for the successful production of spring wheat and barley DHs in Australia.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Abbreviations

BA:

6-Benzyladenine

IAA:

Indole-3-acetic acid

2,4-D:

2,4-Dichlorophenoxyacetic acid

References

  1. Khush GS, Virmani SS (1996) Haploids in plant breeding. In: Jain SR, Sopory SK, Veilleux RE (eds) In vitro haploid production in higher plants, vol 1, Fundamental aspects and methods. Kluwer Academic Publishers, Dordrecht, pp 11–33

    Chapter  Google Scholar 

  2. Touraev A, Pfosser M, Heberle-Bors E (2001) The microspore: a haploid multipurpose cell. Adv Bot Res 35:53–109

    Article  Google Scholar 

  3. Ascher-Ellis JS, Graham RD, Hollamby GJ et al (2001) In: Reynolds MP, Ortiz-Monasterio JI, McNab A (eds) Application of physiology in wheat breeding. CIMMYT, Mexico, DF, pp 219–240

    Google Scholar 

  4. Devaux P, Pickering R (2005) Haploids in the improvement of Poaceae. In: Palmer CE, Keller WA, Kasha KJ (eds) Biotechnology in agriculture and forestry—56 haploids in crop improvement II. Springer, Heidelberg, pp 215–242

    Google Scholar 

  5. Jauhar PP, Xu SS, Baenziger PS (2009) Haploidy in cultivated wheats: induction and utility in basic and applied research. Crop Sci 49:737–755

    Article  Google Scholar 

  6. Collard BCY, Jahurfer MZZ, Brouer JB et al (2005) An introduction to markers, quantitative trait loci (QTL) mapping and marker-assisted selection for crop improvement: the basic concepts. Euphytica 142:169–196

    Article  CAS  Google Scholar 

  7. Kammholz SJ, Campbell AW, Sutherland MW et al (2001) Establishment and characterisation of wheat genetic mapping populations. Aust J Agric Res 52:1079–1088

    Article  CAS  Google Scholar 

  8. Cakir M, Gupta S, Li CD et al (2011) Genetic mapping and QTL analysis of disease resistance traits in the barley population Baudin × AC Metcalfe. Crop Pasture Sci 62:152–161

    Article  Google Scholar 

  9. Howes NK, Woods SM, Townley-Smith TF (1998) Simulations and practical problems of applying marker-assisted selection and doubled haploids to wheat breeding programs. Euphytica 100:225–230

    Article  Google Scholar 

  10. Bonnett DG, Rebetzke GJ, Spielmeyer W (2005) Strategies for efficient implementation of molecular markers in wheat breeding. Mol Breed 15:75–85

    Article  CAS  Google Scholar 

  11. Kuchel H, Ye G, Fox R et al (2005) Genetic and economic analysis of a targeted marker-assisted wheat breeding strategy. Mol Breed 16:67–78

    Article  Google Scholar 

  12. Devaux P, Kasha K (2009) Overview of barley doubled haploid production. In: Touraev A, Forster BP, Jain SM (eds) Advances in haploid production in higher plants. Springer, Heidelberg, pp 47–63

    Chapter  Google Scholar 

  13. Ding XL, Luckett DJ, Darvey NL (1995) A cost based index of anther culture response in diverse wheat-breeding germplasm. Aust J Exp Agric 35:395–401

    Article  Google Scholar 

  14. Broughton S (2008) Ovary co-culture improves embryo and green plant production in anther culture of Australian spring wheat genotypes (Triticum aestivum L.). Plant Cell Tiss Org Cult 95:185–195

    Article  Google Scholar 

  15. Tuvesson S, Ljungberg A, Johansson N et al (2000) Large-scale production of wheat and triticale doubled haploids through the use of a single-anther culture method. Plant Breed 119:455–459

    Article  Google Scholar 

  16. Davies P (2003) Barley isolated microspore culture (IMC) method. In: Malszynnski M, Kasha KJ, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants: a manual. Kluwer Academic Publishers, Dordrecht, pp 29–34

    Google Scholar 

  17. Weyen J (2009) Barley and wheat doubled haploids in breeding. In: Touraev A, Forster BP, Jain SM (eds) Advances in haploid production in higher plants. Springer, Heidelberg, pp 179–187

    Chapter  Google Scholar 

  18. Jacquard C, Wojnarowiez G, Clement C (2003) Anther culture in barley. In: Malszynnski M, Kasha KJ, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants: a manual. Kluwer Academic Publishers, Dordrecht, pp 21–27

    Chapter  Google Scholar 

  19. Broughton S (2011) The application of n-butanol improves embryo and green plant production in anther culture of Australian wheat (Triticum aestivum L.) genotypes. Crop Pasture Sci 62:813–822

    Article  CAS  Google Scholar 

  20. Ouyang J, Liang H, Jia S et al (1994) Studies on the chromosome doubling of wheat pollen plants. Plant Sci 98:209–214

    Article  CAS  Google Scholar 

  21. Navarro-Alvarez W, Baenziger PS, Eskridge KM et al (1994) Addition of colchicine to wheat anther culture media to increase doubled haploid production. Plant Breed 112:192–198

    Article  CAS  Google Scholar 

  22. Barnabás B, Pfahler PL, Kovacs G (1991) Direct effect of colchicine on the microspore embryogenesis to produce dihaploid plants in wheat (Triticum aestivum L.). Theor Appl Genet 81:675–678

    Article  PubMed  Google Scholar 

  23. Hansen NJP, Anderson SB (1998) In vitro chromosome doubling with colchicine during microspore culture in wheat (Triticum aestivum L). Euphytica 102:101–108

    Article  CAS  Google Scholar 

  24. Soriano M, Cistué L, Vallés MP et al (2007) Effects of colchicine on anther and microspore culture of bread wheat (Triticum aestivum L.). Plant Cell Tiss Org Cult 91:225–234

    Article  CAS  Google Scholar 

  25. Weyen J, Hoenemann C (personal communication)

    Google Scholar 

  26. Suenaga K, Nakajima K (1989) Efficient production of haploid wheat T. aestivum through crosses between Japanese wheat and maize Zea mays. Plant Cell Rep 8:263–266

    Article  CAS  PubMed  Google Scholar 

  27. Zenkteler M, Nitzsche W (1984) Wide hybridization experiments in cereals. Theor Appl Genet 68:311–315

    Article  CAS  PubMed  Google Scholar 

  28. Laurie DA, O’Donoughue S, Bennet MD (1990) Wheat x maize and other wide sexual hybrids: their potential for genetic manipulation and crop improvement. In: Gustavson JP (ed) Gene manipulation in plant improvement. Plenum, New York, pp 95–126

    Chapter  Google Scholar 

  29. Suenaga K (1994) Doubled haploid system using the intergeneric crosses between wheat (Triticum aestivum) and maize (Zea mays). Bull Nat Inst Agrobiol Resour 9:83–139

    CAS  Google Scholar 

  30. Sood S, Dhawaan R, Singh K et al (2003) Development of novel chromosome doubling strategies for wheat × maize system of wheat haploid production. Plant Breed 122:493–496

    Article  Google Scholar 

  31. Jensen CJ (1974) Chromosome doubling techniques in haploids. In: Kasha KJ (ed) Haploids in higher plants. Advances and potential. Proceedings of the first international symposium. Guelph University Press, Guelph, pp 153–190

    Google Scholar 

  32. Kasha KJ (2005) Chromosome doubling and the recovery of doubled haploid plants. In: Palmer CE, Keller WA, Kasha KJ (eds) Biotechnology in agriculture and forestry—56 haploids in crop improvement II. Springer, Heidelberg, pp 123–147

    Google Scholar 

  33. Cistué L, Ramos A, Castillo AM et al (1994) Production of large numbers of doubled haploid plants from barley anthers pre-treated with high concentrations of mannitol. Plant Cell Rep 13:709–712

    Article  PubMed  Google Scholar 

  34. Broughton S, Priest P (1997) Barley doubled haploids in Western Australia—improved green plant regeneration from anthers pretreated with high concentrations of mannitol. Eighth Australian barley technical symposium, Gold Coast, Queensland, 7–12 Sep 1997

    Google Scholar 

  35. Hoekstra S, van Bergen S, van Brouwershaven IR et al (1997) Androgenesis in Hordeum vulgare L.: effects of mannitol, calcium and abscisic acid on anther pretreatment. Plant Sci 126:211–218

    Article  CAS  Google Scholar 

  36. Cistué L, Vallés MP, Echávarri B et al (2004) Production of barley doubled haploid by anther and microspore culture. In: Mujib A, Myeong-Je C, Predieri S, Banjaree S (eds) In vitro applications in crop improvements—recent progress. Oxford & IBH Publishing Co. Pvt. Ltd, New Delhi, India, pp 1–18

    Google Scholar 

  37. Hunter CP (1988) Plant regeneration from microspores of barley, Hordeum vulgare L. PhD thesis. Wye College, Univ of London, London

    Google Scholar 

  38. Hu TC, Ziauddin A, Simion E et al (1995) Isolated microspore culture of wheat (Triticum aestivum L.) in a defined media I. Effects of pretreatment, isolation methods, and hormones. In Vitro Cell Dev Biol 31:79–83

    Article  CAS  Google Scholar 

  39. Gamborg OL, Miller RA, Ojima K (1968) Nutritional requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  CAS  PubMed  Google Scholar 

  40. Kasha KJ, Simion E, Miner M et al (2003) Haploid wheat isolated microspore culture protocol. In: Malszynnski M, Kasha KJ, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants: a manual. Kluwer Academic Publishers, Dordrecht, pp 77–82

    Chapter  Google Scholar 

  41. Soriano M, Cistué L, Castillo AM (2008) Enhanced induction of microspore embryogenesis after n-butanol treatment in wheat (Triticum aestivum L.) anther culture. Plant Cell Rep 27:805–811

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Agriculture and Food, Government of WA, 3 Baron-Hay Court, South Perth, 6151, WA, Australia

    Sue Broughton

  2. South Australian Research and Development Institute, Adelaide, SA, Australia

    Parminder K. Sidhu & Philip A. Davies

Authors
  1. Sue Broughton
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Parminder K. Sidhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Philip A. Davies
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sue Broughton .

Editor information

Editors and Affiliations

  1. University of Adelaide, Australian Centre for Plant Functional Genomics (ACPFG), Urrbrae, South Australia, Australia

    Delphine Fleury

  2. University of Adelaide, Australian Centre for Plant Functional Genomics (ACPFG), Urrbrae, South Australia, Australia

    Ryan Whitford

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Broughton, S., Sidhu, P.K., Davies, P.A. (2014). In vitro Culture for Doubled Haploids: Tools for Molecular Breeding. In: Fleury, D., Whitford, R. (eds) Crop Breeding. Methods in Molecular Biology, vol 1145. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0446-4_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-0446-4_14

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0445-7

  • Online ISBN: 978-1-4939-0446-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation