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The auxins centrophenoxine and 2,4-D differ in their effects on non-directly induced chromosome doubling in anther culture of wheat (T. aestivum L.)

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Abstract

Doubled haploid technologies have become key tools for plant breeding. Using these techniques, the speed and efficiency of plant improvement processes can be significantly enhanced. Anther culture-based technologies have the potential to regenerate large numbers of doubled haploid plants without colchicine treatment. In an attempt to elucidate the influence of phytohormones on non-directly induced chromosome doubling, two synthetic auxins, 2,4-D and centrophenoxine, were tested in a wheat anther culture approach. Whereas the induction of androgenic embryo-like structures (ELSs) was efficient for both auxins, we observed a significantly higher frequency of chromosome doubling when using 2,4-D than when using centrophenoxine. When 2,4-D was added to the induction medium, a positive correlation between the size of ELSs and their ploidy level was detected by flow cytometry. The morphological selection of ELSs, a process that was included in routine operations of the method without significantly extending the input of time and effort, facilitates the production of fertile DH plants with a frequency of 60 %. Our findings may contribute to a more efficient production of doubled haploid wheat plants using a colchicine-free anther culture approach.

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References

  • Armstrong T, Metz S, Mascia P (1987) Two regeneration systems for the production of haploid plants from wheat anther culture. Plant Sci 51:231–237

    Article  Google Scholar 

  • Barnabás B (2003) Protocol for producing doubled haploid plants from anther culture of wheat (Triticum aestivum L.). In: Maluszynski M, Kasha K, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants: a manual. Kluwer, Dordrecht, pp 65–70

    Chapter  Google Scholar 

  • Castillo AM, Cistué L, Vallés MP, Soriano M (2009) Chromosome doubling in monocots. In: Touraev A, Forster BP, Jain SM (eds) Advances in haploid production in higher plants. Springer, Netherlands, pp 329–338

    Chapter  Google Scholar 

  • Dubas E, Wędzony M, Petrovska B, Salaj J, Żur I (2010) Cell structural reorganization during induction of androgenesis in isolated microspore cultures of triticale (× Triticosecale Wittm.). Acta Biol Cracoviensia Ser Bot 52:73–86

    Google Scholar 

  • Ferrie AMR, Caswell KL (2011) Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production. Plant Cell, Tissue Organ Cult 104:301–309

    Article  Google Scholar 

  • Forster BP, Heberle-Bors E, Kasha KJ, Touraev A (2007) The resurgence of haploids in higher plants. Trends Plant Sci 12:368–375

    Article  PubMed  CAS  Google Scholar 

  • Germana MA (2011) Anther culture for haploid and doubled haploid production. Plant Cell, Tissue Organ Cult 104:283–300

    Article  Google Scholar 

  • Grasser M, Kane CM, Merkle T, Melzer M, Emmersen J, Grasser KD (2009) Transcript elongation factor TFIIS is involved in Arabidopsis seed dormancy. J Mol Biol 386:598–611

    Article  PubMed  CAS  Google Scholar 

  • Grauda D, Lepse N, Strazdiņa V, Kokina I, Lapiņa L, Miķelsone A, Ļubinskis L, Rashal I (2010) Obtaining of doubled haploid lines by anther culture method for the Latvian wheat breeding. Agron Res 8:545–552

    Google Scholar 

  • Hoekstra S, van Zijderveld M, Heidekamp E, van der Mark E (1993) Microspore culture of Hordeum vulgate L.: the influence of density and osmolality. Plant Cell Rep 12:331–335

    Article  Google Scholar 

  • Inagaki MN (2003) Doubled haploid production in wheat using wide hybridization. In: Maluszynski M, Kasha K, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants: a manual. Kluwer, Dordrecht, pp 53–58

    Chapter  Google Scholar 

  • Indrianto A, Barinova I, Touraev A, Heberle-Bors E (2001) Tracking individual wheat microspores in vitro: identification of embryogenic microspores and body axis formation in the embryo. Planta 212:163–174

    Article  PubMed  CAS  Google Scholar 

  • Kahrizi D (2009) Study of androgenesis and spontaneous chromosome doubling in barley (Hordeum vulgare L.). Advanced lines using isolated microspore culture. Int J Plant Breed 3:111–114

    Google Scholar 

  • Kahrizi D, Mohammadi R (2009) Study of androgenesis and spontaneous chromosome doubling in barley (Hordeum vulgare L.) genotypes using isolated microspore culture. Acta Agron Hung 52:155–164

    Article  Google Scholar 

  • Kahrizi D, Zebarjadi AR, Jalali Honarmand S, Khah EM, Motamedi J (2009) Study of spontaneous chromosome doubling in wheat (Triticum aestivum L.) genotypes using androgenesis. World Acad Sci Eng Technol 57:1162–1166

    Google Scholar 

  • Kahrizi D, Mahmoodi S, Bakhshi Khaniki GR, Mirzaei M (2011) Effect of genotype on androgenesis in barley (Hordeum vulgare L.). Biharean Biol 5:132–134

    Google Scholar 

  • Kasha KJ, Simion E, Miner M, Letarte J, Hu TC (2003) Haploid wheat isolated microspore culture protocol. In: Maluszynski M, Kasha K, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants: a manual. Kluwer, Dordrecht, pp 77–81

    Chapter  Google Scholar 

  • Kempe K, Rubtsova M, Berger C, Kumlehn J, Schollmeier C, Gils M (2010) Transgene excision from wheat chromosomes by phage phiC31 integrase. Plant Mol Biol 72:673–687

    Article  PubMed  CAS  Google Scholar 

  • Kudirka DT, Schaeffer G, Baenziger P (1989) Stability of ploidy in meristems of plants regenerated from anther calli of wheat (Triticum aestivum L. em. Thell.). Genome 32(6):1068–1073

    Article  Google Scholar 

  • Lantos C, Páricsi S, Zofajova A, Weyen J, Pauk J (2006) Isolated microspore culture of wheat (Triticum aestivum L.) with Hungarian cultivars. Acta Biol Szeged 50(1–2):31–35

    Google Scholar 

  • Little E, Magbanua Z, Parrott W (2000) A protocol for repetitive somatic embryogenesis from mature peanut epicotyls. Plant Cell Rep 19:351–357

    Article  CAS  Google Scholar 

  • Lolas IB, Himanen K, Gronlund JT, Lynggaard C, Houben A, Melzer M, Van Lijsebettens M, Grasser KD (2010) The transcript elongation factor FACT affects Arabidopsis vegetative and reproductive development and genetically interacts with HUB1/2. Plant J 61:686–697

    Article  PubMed  CAS  Google Scholar 

  • Maraschin SF, de Priester W, Spaink HP, Wang M (2005) Androgenic switch: an example of plant embryogenesis from the male gametophyte perspective. J Exp Bot 56:1711–1726

    Article  PubMed  CAS  Google Scholar 

  • Muranty H, Sourdille P, Bernard S, Bernard M (2002) Genetic characterization of spontaneous diploid androgenetic wheat and triticale plants. Plant Breed 121:470–474

    Article  CAS  Google Scholar 

  • Niroula RK, Bimb HP (2009) Overview of wheat X maize system of crosses for dihaploid induction in wheat. World Appl Sci J 7:1037–1045

    CAS  Google Scholar 

  • Pauk J, Miháli R, Puolimatka M (2003) Protocol of wheat (Triticum aestivum L.) anther culture. In: Maluszynski M, Kasha K, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants: a manual. Kluwer, Dordrecht, pp 59–64

    Chapter  Google Scholar 

  • Schlegel R, Belchev I, Kostov K, Atanasova M (2000) Inheritance of high anther culture response in hexaploid wheat, Triticum aestivum L. var. Svilena. Bulg J Agric Sci 6(3):261–270

    Google Scholar 

  • Shariatpanahi ME, Belogradova K, Hessamvaziri L, Heberle-Bors E, Touraev A (2006) Efficient embryogenesis and regeneration in freshly isolated and cultured wheat (Triticum aestivum L.) microspores without stress pretreatment. Plant Cell Rep 25:1294–1299

    Article  PubMed  CAS  Google Scholar 

  • Steinitz B, Küsek M, Tabib Y, Paran I, Zelcer A (2003) Pepper (Capsicum annuum L.) regenerants obtained by direct somatic embryogenesis fail to develop a shoot. In Vitro Cell Dev Biol 39:296–303

    CAS  Google Scholar 

  • Stober A, Hess D (1997) Spike pretreatments, anther culture conditions, and anther culture response of 17 German varieties of spring wheat (Triticum aestivum L.). Plant Breed 116:443–447

    Article  Google Scholar 

  • Touraev A, Indrianto A, Wratschko I, Vicente O, Heberle-Bors E (1996) Efficient microspore embryogenesis in wheat (Triticum aestivum L.) induced by starvation at high temperature. Sex Plant Reprod 9:209–215

    Article  Google Scholar 

  • Wędzony M, Forster BP, Żur I, Golemiec E, Szechyńska-Hebda M, Dubas E, Gotębiowska G (2009) Progress in doubled haploid technology in higher plants. In: Touraev A, Forster BP, Jain SM (eds) Advances in haploid production in higher plants. Springer, Netherlands, pp 1–34

  • 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, Netherlands, pp 179–187

    Chapter  Google Scholar 

  • Ziauddin A, Kasha KJ (1989) Long-term callus cultures of diploid barley (Hordeum vulgare). II. Effect of auxins on chromosomal status of cultures and regeneration of plants. Euphytica 48:279–286

    Article  Google Scholar 

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Acknowledgments

We thank Corinna Schollmeier for outstanding technical assistance and greenhouse management. We are especially grateful to Karla Anders and Sandra Bössow for excellent technical assistance in donor plant production and tissue culture work. We thank Corine Graser, Silvana Fischer, Josephine Tubis, Kerstin Denzin, Monika Wiesner, Kirsten Hoffie and Twan Rutten for technical support. In particular, the authors would like to express their gratitude to Wolf v. Rhade and Ralf Schachschneider (Nordsaat GmbH, Langenstein, Germany) for their permanent support. The research was financed by the Nordsaat GmbH, Böhnshausen and the IPK-Gatersleben.

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Authors and Affiliations

  1. Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK) Gatersleben, Corrensstraße 3, 06466, Stadt Seeland OT Gatersleben, Germany

    Myroslava Rubtsova, Michael Melzer & Mario Gils

  2. Saaten-Union Biotec GmbH, Am Schwabeplan 6, 06466, Stadt Seeland OT Gatersleben, Germany

    Heike Gnad

  3. Saaten-Union Biotec GmbH, Hovedisser Str. 92, 33818, Leopoldshoehe, Germany

    Jens Weyen

Authors
  1. Myroslava Rubtsova
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  2. Heike Gnad
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  3. Michael Melzer
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  4. Jens Weyen
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  5. Mario Gils
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Corresponding author

Correspondence to Mario Gils.

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M. Rubtsova and H. Gnad contributed equally to the paper.

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Rubtsova, M., Gnad, H., Melzer, M. et al. The auxins centrophenoxine and 2,4-D differ in their effects on non-directly induced chromosome doubling in anther culture of wheat (T. aestivum L.). Plant Biotechnol Rep 7, 247–255 (2013). https://doi.org/10.1007/s11816-012-0256-x

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  • DOI: https://doi.org/10.1007/s11816-012-0256-x

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