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Doubled Haploid Onion (Allium cepa L.) Production Via In Vitro Gynogenesis

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Doubled Haploid Technology

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

Abstract

Onion (A. cepa L.) is an outcrossing biennial species with a very large genome. Development of genetically uniform (inbred) lines highly desired by onion breeders is a difficult process due to high level of heterozygosity. Inbred onion development may take up to five generations (~10 years) by classical selfing technique. Onion shows severe inbreeding depression, which further complicates production of lines with stabilized important agronomic traits. When applied successfully, haploidization technology can be useful in the development of fully homozygous onion lines in 2 years. Although production of haploid and doubled haploid (DH) onions via gynogenesis was reported more than three decades ago, successful production and utilization of DHs in onion breeding is still far behind of expectations of breeders. The main obstacles in front of the success include high variation in the response of donor materials to gynogenesis induction and difficulties faced in the process of obtaining DHs from haploid plants. We use a DH production procedure enabling us to develop DH plants from a wide range of onion donor materials. This procedure is based on production of haploid plants via single step culture of unopened flower buds, detection of haploid plants among gynogenic regenerants, and converting these plants to fecund DHs using a combination of ploidy manipulation techniques. The bulbs of DHs are produced in about 1 year after the initiation of induction cultures and selfed seeds are produced from fecund DH plants when they flower in the second year.

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References

  1. Brewster JL (2008) Onions and other vegetable alliums, 2nd edn. CAB International, Wallingford, UK

    Book  Google Scholar 

  2. Hanelt P (1990) Taxonomy, evolution and history. In: Rabinowitch HD, Brewster JL (eds) Onions and allied crops. botany, physiology, and genetics, vol I. CRC Press, Boca Raton, Florida, pp 1–26

    Google Scholar 

  3. Fritsch RM, Friesen N (2002) Evolution, domestication and taxonomy. In: Rabinowitch HD, Currah L (eds) Allium crop science: recent advances. CABI Publishing, New York, pp 5–30

    Chapter  Google Scholar 

  4. FAO. (2017). FAOSTAT. Food and Agriculture Organization of the United Nations FAO Statistical Database. http://faostat.fao.org/

  5. Bijl JR (1994) Allium-flowering onions. Herbertia 50:88–94

    Google Scholar 

  6. Kamenetsky R, Frisch RM (2002) Ornamental Alliums. In: Rabinowitch HD, Brewster JL (eds) Allium crop science: recent advances. CABI Publishing, New York, pp 459–491

    Chapter  Google Scholar 

  7. Huber LS, Hoffmann-Ribani R, Rodriguez-Amaya DB (2009) Quantitative variation in Brazilian vegetable sources of flavonols and flavones. Food Chem 113:1278–1282

    Article  CAS  Google Scholar 

  8. Zill-E-Huma, Vian MA, Maingonnat JF, Chemat F (2009) Clean recovery of antioxidant flavonoids from onions: Optimising solvent free microwave extraction method. J Chromatogr A 1216:7700–7707

    Article  CAS  PubMed  Google Scholar 

  9. Keller ERJ, Korzun L (1996) Haploidy in onion (Allium cepa L.) and other allium species. In: Jain SM, Sopory SK, Veilleux RE (eds) In vitro haploid production in higher plants, vol 3. Kluwer, Dordrecht, pp 51–71

    Chapter  Google Scholar 

  10. Muren RC (1989) Haploid plant induction from unpollinated ovaries in onion. Hortic Sci 24:833–834

    Google Scholar 

  11. Keller J (1990) Culture of unpollinated ovules, ovaries, and fower buds in some species of the genus allium and haploid induction via gynogenesis in onion (Allium cepa L.). Euphytica 47:241–247

    Google Scholar 

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

    Article  CAS  Google Scholar 

  13. Dore C, Marie F (1993) Production of gynogenetic plants of onion (Allium cepa L.) after crossing with irradiated pollen. Plant Breed 111:142–147

    Article  Google Scholar 

  14. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  16. Dunstan DI, Short KC (1977) Improved growth of tissue cultures of onion, Allium cepa. Physiol Plant 41:70–72

    Article  Google Scholar 

  17. Campion B, Azzimonti MT, Vicini E, Schiavi M, Falavigna A (1992) Advances in haploid plant induction in onion (Allium cepa L.) through in vitro gynogenesis. Plant Sci 86:97–104

    Article  Google Scholar 

  18. Bohanec B, Jakse M, Ihan A, Javornik B (1995) Studies of gynogenesis in onion (Allium cepa L.) induction procedures and genetic analysis of regenerants. Plant Sci 104:215–224

    Article  CAS  Google Scholar 

  19. Geoffriau E, Kahane R, Rancillac M (1997) Variation of gynogenesis ability in onion (Allium cepa L.). Euphytica 94:37–44

    Article  Google Scholar 

  20. Martinez LE, Aguero CB, Lopez ME, Galmarini CR (2000) Improvement of in vitro gynogenesis induction in onion (Allium cepa L.) using polyamines. Plant Sci 156:221–226

    Article  CAS  PubMed  Google Scholar 

  21. Michalik B, Adamus A, Nowak E (2000) Gynogenesis in polish onion cultivars. J Plant Physiol 156:211–216

    Article  CAS  Google Scholar 

  22. Alan AR, Mutschler MA, Brants A, Cobb E, Earle ED (2003) Production of gynogenic plants from hybrids of Allium cepa L. and A. roylei Stearn. Plant Sci 165:1201–1211

    Article  CAS  Google Scholar 

  23. Alan AR, Brants A, Cobb E, Goldschmied PA, Mutschler MA, Earle ED (2004) Fecund gynogenic lines from onion (Allium cepa L.) breeding materials. Plant Sci 167:1055–1066

    Article  CAS  Google Scholar 

  24. Celebi-Toprak F, Kaska A, Alan V, Aykut A, Alan AR (2015) Evaluation of doubled haploid onion (Allium cepa L.) lines developed from Turkish landraces. In: ISEA 2015|7th International Symposium on Edible Alliaceae 21st–25th May 2015. Nigde, Turkey, p 81

    Google Scholar 

  25. Fayos O, Valles MP, Garces-Claver A, Mallor C, Castillo AM (2015) Doubled haploid production from Spanish onion (Allium cepa L.) germplasm: embryogenesis induction, plant regeneration and chromosome doubling. Front Plant Sci 6:384. https://doi.org/10.3389/fpls.2015.00384

    Article  PubMed  PubMed Central  Google Scholar 

  26. Khar A, Kumar A, Islam S, Kumar A, Agarwal A (2018) Genotypic response towards haploid induction in short day tropical Indian onion (Allium cepa L.). Ind J Agric Sci 88:709–713

    CAS  Google Scholar 

  27. Alan AR, Lim W, Mutschler MA, Earle ED (2007) Complementary strategies for ploidy manipulations in gynogenic onion (Allium cepa L.). Plant Sci 173:25–31

    Article  CAS  Google Scholar 

  28. Geoffriau E, Kahane R, Bellamy C, Rancillac M (1997) Ploidy stability and in vitro chromosome doubling in gynogenic clones of onion (Allium cepa L.). Plant Sci 122:201–208

    Article  CAS  Google Scholar 

  29. Jakse M, Havey MJ, Bohanec B (2003) Chromosome doubling procedures of onion (Allium cepa L.) gynogenic embryos. Plant Cell Rep 21:905–910

    Article  CAS  PubMed  Google Scholar 

  30. Grzebelus E, Adamus A (2004) Efect of anti-mitotic agents on development and genome doubling of gynogenic onion (Allium cepa L.) embryos. Plant Sci 167:569–574

    Article  CAS  Google Scholar 

  31. Khosa JS, Lee R, Brauning S, Lord J, Pither-Joyce M, McCallum J, Macknight RC (2016) Doubled haploid ‘CUDH2107’ as a reference for bulb onion (Allium cepa L.) research: development of a transcriptome catalogue and identification of transcripts associated with male fertility. PLoS One 11:e166568

    Google Scholar 

  32. Finkers R, Van Workum W, Van Kaauwen M, Huits H, Jungerius A, Vosman B, Scholten OE (2015) A de novo assembly for the 16 GB Allium cepa genome, tears of joy. San Diego: Plant & Animal Genome XXIII; 2015. https://pag.confex.com/pag/xxiii/webprogram/Paper17794.html

  33. Hyde PT, Earle ED, Mutschler MA (2012) Doubled haploid onion (Allium cepa L.) lines and their impact on hybrid performance. Hortic Sci 47:1690–1695

    Google Scholar 

  34. Peterson R, Slovin JP, Chen C (2010) A simplified method for differential staining of aborted and non-aborted pollen grains. Int J Plant Biol 1:e13

    Article  Google Scholar 

  35. Arumuganathan K, Earle ED (1991) Nuclear DNA content of some important plant species. Plant Mol Biol Report 9(3):211–215

    Google Scholar 

  36. Dolezel J, Bartos J (2005) Plant DNA flow cytometry and estimation of genome size. Ann Bot 95:99–110

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

This research was supported by grants from The Scientific and Technological Research Council of Turkey (TUBITAK-TOVAG, Project Nos. 110O095 and 117O386). Special thanks to members of PAU BIYOM (Pamukkale University, Denizli, Turkey) and Earle Lab (Cornell University, Ithaca, NY, USA).

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Authors and Affiliations

  1. Plant Genetics and Agricultural Biotechnology Application and Research Center (PAU BIYOM), Pamukkale University, Denizli, Turkey

    Ali Ramazan Alan

  2. Department of Biology, Pamukkale University, Denizli, Turkey

    Ali Ramazan Alan

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  1. Ali Ramazan Alan
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Correspondence to Ali Ramazan Alan .

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Editors and Affiliations

  1. CPI, COMAV - Univ Politecnica Valencia, Valencia, Valencia, Spain

    Jose M. Segui-Simarro

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Alan, A.R. (2021). Doubled Haploid Onion (Allium cepa L.) Production Via In Vitro Gynogenesis. 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_6

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  • DOI: https://doi.org/10.1007/978-1-0716-1315-3_6

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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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