Advertisement

Barley pp 37-52 | Cite as

Barley Anther Culture

  • Ludmila Ohnoutkova
  • Tomas Vlcko
  • Mentewab Ayalew
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1900)

Abstract

The production of doubled haploid (DH) barley plants through anther culture is a very useful yet simple in vitro technique. DH plants derive from divisions of haploid microspores that have undergone a developmental switch under the appropriate conditions. The successive divisions lead to the formation of an embryo or callus rather than the formation of mature pollen grains. Plants that regenerate from these embryos are often either haploid, in which case their chromosome set can be doubled by treatment with colchicine, or spontaneous double haploids. The efficiency of DH plant production is highly variable depending on the genotype of the source material. Despite this limitation, DH plants have been widely used in breeding and research programs. Compared to conventional approaches, breeding strategies that makes use of DH plants achieve a homozygous state, allowing transgene or mutation stabilization in the genome, within a considerably shorter time, thus accelerating workflow or reducing work volume.

Key words

Barley (Hordeum vulgareAnther culture Doubled haploid Androgenesis 

Notes

Acknowledgments

This work was supported by the Ministry of Education, Youth and Sports, Czech Republic (Grant LO1204 from the National Program of Sustainability I.). We would like to thank the Gordon-Zeto Center for Global Education at Spelman College for supporting Tomas Vlcko and thus facilitating our collaboration. The authors are grateful to Ota Blahousek for the technical assistance.

References

  1. 1.
    Guha S, Maheshwari SC (1964) In vitro production of embryos from anthers of Datura. Nature 204:497CrossRefGoogle Scholar
  2. 2.
    Bourgin JP, Nitsch JP (1967) Production of haploids nicotiana from excised stamens. Annales De Physiologie Vegetale 9:377–382Google Scholar
  3. 3.
    Clapham D (1973) Haploid Hordeum plants from anthers in-vitro. J Plant Breed 69:142–155Google Scholar
  4. 4.
    Hu H (1997) In vitro induced haploids in wheat. In: Jain SM, Sopory SK, Veilleux RE (eds) In vitro haploid production in higher plants: Volume 4—Cereals. Springer Netherlands, Dordrecht, pp 73–97CrossRefGoogle Scholar
  5. 5.
    Li H, Devaux P (2005) Isolated microspore culture overperforms anther culture for green plant regeneration in barley (Hordeum vulgare L.). Acta Physiol Plant 27:611–619CrossRefGoogle Scholar
  6. 6.
    Lantos C, Páricsi S, Zofajova A et al (2006) Isolated microspore culture of wheat (Triticum aestivum L.) with Hungarian cultivars. Acta Biologica Szegediensis 50:31–35Google Scholar
  7. 7.
    Reynolds TL (1997) Pollen embryogenesis. Plant Mol Biol 33:1–10CrossRefGoogle Scholar
  8. 8.
    Bonet FJ, Azbaid L, Olmedilla A (1998) Pollen embryogenesis: atavism or totipotency? Protoplasma 202:115–121CrossRefGoogle Scholar
  9. 9.
    Sunderland N, Evans LJ (1980) Multicellular pollen formation in cultured barley anthers. J Exp Bot 31:501–514CrossRefGoogle Scholar
  10. 10.
    Chen X-W, Cistué L, Muñoz-Amatriaín M et al (2007) Genetic markers for doubled haploid response in barley. Euphytica 158:287–294CrossRefGoogle Scholar
  11. 11.
    Lazaridou T, Sistanis I, Lithourgidis A et al (2011) Response to in-vitro anther culture of F 3 families originating from high and low yielding F 2 barley (Hordeum vulgare L.) plants. Aust J Crop Sci 5:265–270Google Scholar
  12. 12.
    Jacquard C, Nolin F, Hécart C et al (2009) Microspore embryogenesis and programmed cell death in barley: effects of copper on albinism in recalcitrant cultivars. Plant Cell Rep 28:1329–1339CrossRefGoogle Scholar
  13. 13.
    Makowska K, Oleszczuk S, Zimny A et al (2015) Androgenic capability among genotypes of winter and spring barley. Plant Breed 134:668–674CrossRefGoogle Scholar
  14. 14.
    Rodríguez-Serrano M, Bárány I, Prem D et al (2012) NO, ROS, and cell death associated with caspase-like activity increase in stress-induced microspore embryogenesis of barley. J Exp Bot 63:2007–2024CrossRefGoogle Scholar
  15. 15.
    Caredda S, Doncoeur C, Devaux P et al (2000) Plastid differentiation during androgenesis in albino and non-albino producing cultivars of barley (Hordeum vulgare L.). Sex Plant Reprod 13:95–104CrossRefGoogle Scholar
  16. 16.
    Devaux P, Pickering R (2005) Haploids in the improvement of Poaceae. In: Don Palmer CE, Keller WA, Kasha KJ (eds) Haploids in crop improvement II. Springer Berlin Heidelberg, Berlin, Heidelberg, pp 215–242CrossRefGoogle Scholar
  17. 17.
    Kahrizi D, Mahmoodi S, Khaniki GB et al (2011) Effect of genotype on androgenesis in barley (Hordeum vulgare L.). Biharean Biol 5:132–134Google Scholar
  18. 18.
    Shen Y, Pan G, and Lübberstedt T (2015) Haploid Strategies for Functional Validation of Plant Genes. Trends in Biotechnology. 33, 611–620CrossRefGoogle Scholar
  19. 19.
    Chu CC (1978) The N6 medium and its applications to anther culture of cereal crops. In: Proceedings of symposium on plant tissue culture, Science Press, Peking, p 45–50Google Scholar
  20. 20.
    Zhuang J, Xu J (1983) Increasing differentiation frequencies in wheat pollen callus. In: Hu H, Vega M (eds) Cell and tissue culture techniques for cereal crop improvement. Science Press, Beijing, p 431Google Scholar
  21. 21.
    Dolezel J, Greilhuber J, Suda J (2007) Estimation of nuclear DNA content in plants using flow cytometry. Nat Protocol 2:2233–2244CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Ludmila Ohnoutkova
    • 1
  • Tomas Vlcko
    • 1
  • Mentewab Ayalew
    • 2
  1. 1.Faculty of Science, Centre of the Region Hana for Biotechnological and Agricultural Research, Department of Chemical Biology and GeneticsPalacky UniversityOlomoucCzech Republic
  2. 2.Biology DepartmentSpelman CollegeAtlantaUSA

Personalised recommendations