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Unpollinated Ovaries Used to Produce Doubled Haploid Lines in Durum Wheat

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Doubled Haploid Technology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2287))

Abstract

The use of doubled haploid lines improves the efficiency of cultivar development and homozygous genotypes can be obtained in one generation, as opposed to conventional line production, which requires several cycles of self-pollination. However, in durum wheat (Triticum turgidum subsp. durum Desf.), the low efficiency of green plant regeneration and the very high frequency of albino plants hinder the application of this technique.

We observed the success of using gynogenesis for durum wheat and the significant influence of growing conditions on ovary and callus development, and on plant regeneration. Our results suggested that the cold pretreatment for 2 weeks is efficient for durum wheat. Furthermore, the addition of 2,4-D, vitamins and glutamine, and the use of maltose as sugar source in media improved the ovary regeneration. We describe in this work an efficient method to regenerate green plants from in vitro durum wheat gynogenesis .

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References

  1. Zheng MY, Liu W, Polle E et al (2001) Culture of freshly isolated wheat (Triticum aestivum L.) microspores treated with inducer chemicals. Plant Cell Rep 20:685–690

    Article  CAS  Google Scholar 

  2. Meenakshi S, Nii A, Dipak KS et al (2012) An improved wheat microspore culture technique for the production of doubled haploid plants. Crop Sci. https://doi.org/10.2135/cropsci2012.03.0141

  3. Lu R, Chen Z, Gao R et al (2016) Genotypes-independent optimization of nitrogen supply for isolated microspore cultures in barley. Biomed Res Int. https://doi.org/10.1155/2016/1801646

  4. Soriano MC, Cistué L, Valles MP et al (2007) Effects of colchicine on anther and microspore culture of bread wheat (Triticum aestivum L.). Plant Cell Tissue Org Cult 91:225–234. https://doi.org/10.1007/s11240-007-9288-2

    Article  CAS  Google Scholar 

  5. Castillo AM, Sanchez-Diaz RA, Vallés MP (2015) Effect of ovary induction on bread wheat anther culture: ovary genotype and developmental stage, and candidate gene association. Front Plant Sci 6:1–12

    Article  Google Scholar 

  6. Lantos C, Pauk J (2016) Anther culture as an effective tool in winter wheat (Triticum aestivum L.) breeding. Russ J Genet 52:794–801. https://doi.org/10.1134/S102279541608007X

    Article  CAS  Google Scholar 

  7. Picard E, Crambers E, Mihamou-Ziyyat A (1994) L’haplodiploïdisation: un outil multi-usage pour la génétique et l’amélioration des céréales. In: Aupelf-uref (ed) Quel avenir pour l’amélioration des plantes? John Libbey Eurotext, Paris

    Google Scholar 

  8. Cistué L, Romagosa I, Batlle F et al (2009) Improvements in the production of doubled haploids in durum wheat (Triticum turgidum L.) through isolated microspore culture. Plant Cell Rep 28:727–735

    Article  Google Scholar 

  9. Slama-Ayed O, Bouhaouel I, Ayed S et al (2019) Efficiency of three haplomethods in durum wheat (Triticum turgidum subsp. durum Desf.): isolated microspore culture, gynogenesis and wheat × maize crosses. Czech J Genet Plant Breed 55(3):101–109

    Article  CAS  Google Scholar 

  10. Sibi ML, Kobaissi A, Shekafandeh A (2001) Green haploid plants from unpollinated ovary culture in tetraploid wheat (Triticum durum Desf.). Euphytica 122:351–359

    Article  Google Scholar 

  11. Campion B, Alloni C (1990) Induction of haploid plants in onion (Allium cepa L.) by in vitro culture of unpollinated ovaries. Plant Cell Tissue Org Cult 20:1–6

    Article  CAS  Google Scholar 

  12. Gürel S, Gürel E, Kaya Z (2000) Doubled haploid plant production from unpollinated ovules of sugar beet (Beta vulgaris L.). Plant Cell Rep 19:1155–1159

    Article  Google Scholar 

  13. Slama-Ayed O, Slim-Amara H (2007) Production of doubled haploids in durum wheat (Triticum durum Desf.) through culture of unpollinated ovaries. Plant Cell Tissue Org Cult 91:125–133

    Article  Google Scholar 

  14. Hadziabdic D, Wadt PA, Reed SM (2018) Haploid cultures. In: Trigiano RN, Gray DJ (eds) Plant tissue culture, development and biotechnology, 1st edn. CRC Press, Boca Raton, Florida

    Google Scholar 

  15. Kasha KJ, Simion E, Oro R et al (2001) An improved in vitro technique for isolated microspore culture of barley. Euphytica 120:379–385

    Article  Google Scholar 

  16. Jahier J, Chevre AM, Eber F et al (1992) Techniques de cytogénétique végétale. In: Jahier (ed) . INRA, France

    Google Scholar 

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Acknowledgments

The authors would like to thank the Laboratory of Genetics and Cereal Breeding of National Agronomic Institute of Tunisia (INAT) for his financial support.

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Ayed Slama, O., Slim Amara, H. (2021). Unpollinated Ovaries Used to Produce Doubled Haploid Lines in Durum Wheat. In: Segui-Simarro, J.M. (eds) Doubled Haploid Technology. Methods in Molecular Biology, vol 2287. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1315-3_12

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  • DOI: https://doi.org/10.1007/978-1-0716-1315-3_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1314-6

  • Online ISBN: 978-1-0716-1315-3

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