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Doubled Haploid Laboratory Protocol for Wheat Using Wheat–Maize Wide Hybridization

  • Meenakshi Santra
  • Hong Wang
  • Scott Seifert
  • Scott Haley
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1679)

Abstract

In traditional wheat breeding, the uniformity of lines derived from a breeding population is obtained by repeated selfing from the F1 which takes several generations to reach homozygosity in loci controlling traits of interest. Using doubled haploid technology, however, it is possible to attain 100% homozygosity at all loci in a single generation and completely homogeneous breeding lines can be obtained in 1–2 years. Thus, doubled haploid technology may significantly reduce cultivar development time. Two major methods for producing wheat doubled haploids are androgenesis (anther culture and microspore culture) and embryo culture using wheat–maize wide hybridization, the latter being the most effective and widely used method. The method of wide hybridization between wheat and maize is laborious but is widely successful for rapidly obtaining homozygous lines. This technique includes six major steps: emasculation of the wheat flower; pollination of the emasculated flower with maize pollen; hormone treatment; embryo rescue; haploid plant regeneration in tissue culture medium; and chromosome doubling. It has been observed that the efficiency of doubled haploid production depends on both maize and wheat genotypes, good plant health and proper greenhouse conditions (without disease, insects, or drought stress), and proper conduct of all procedures. Therefore, the procedures may need minor modification in order to produce higher numbers of embryos, haploid green plants, and doubled haploid plants.

Key words

Wheat Doubled haploid Homozygous Wide hybridization Embryo rescue 

References

  1. 1.
    Maluszynski M, Kasha KJ, Szarejko I (2003) Published doubled haploid protocols in plant species. In: Maluszynski M et al (eds) Doubled haploid production in crop plants. Kluwer Academic Publishers, Dordrecht, pp 309–335CrossRefGoogle Scholar
  2. 2.
    Forster BP, Thomas WTB (2005) Doubled haploids in genetics and plant breeding. In: Janick J (ed) Plant breeding reviews, vol 25. John Wiley & Sons, Inc., Hoboken, NJ, pp 57–88Google Scholar
  3. 3.
    Niu Z, Jiang A, Hammad WA et al (2014) Review of doubled haploid production in durum and common through wheat x maize hybridization. Plant Breed 133:313–320CrossRefGoogle Scholar
  4. 4.
    Barclay IR (1975) High frequencies of haploid production in wheat (Triticum aestivum) by chromosome elimination. Nature 256:410–411CrossRefGoogle Scholar
  5. 5.
    Laurie DA, Bennet MD (1986) Wheat x maize hybridization. Can J Genet Cytol 28:313–316CrossRefGoogle Scholar
  6. 6.
    Suenaga K, Nakajima K (1989) Efficient production of haploid wheat (Triticum aestivum) through crosses between Japanese wheat and maize (Zea mays). Plant Cell Rep 8:263–266CrossRefPubMedGoogle Scholar
  7. 7.
    Inagaki MN (1990) Wheat haploids through the bulbosum technique. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry. Springer, Berlin, pp 448–459Google Scholar
  8. 8.
    Inagaki MN, Nagamine T, Mujeeb-Kazi A (1997) Use of pollen storage and detached-tiller culture in wheat polyhaploid production through wide crosses. Cereal Res Commun 25(1):7–13Google Scholar
  9. 9.
    Murashige T, Skoog S (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Meenakshi Santra
    • 1
  • Hong Wang
    • 1
  • Scott Seifert
    • 1
  • Scott Haley
    • 1
  1. 1.Department of Soil and Crop ScienceColorado State UniversityFort CollinsUSA

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