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Culture and Time-Lapse Tracking of Barley Microspore-Derived Embryos

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Part of the book series: Methods In Molecular Biology™ ((MIMB,volume 427))

Summary

Barley microspore embryogenesis represents an attractive system to study stress-induced cell differentiation and is a valuable tool for efficient plant breeding. In contrast to zygotic embryogenesis, all developmental stages are freely accessible at a large scale for observation, molecular analysis and manipulation techniques. In barley, there is a high percentage of microspores that become embryogenic after stress treatment in a mannitol solution. These microspores have the capacity to follow an embryogenic route in both liquid and solid cultures, yielding up to 10% of embryos. In this protocol, we describe three different culture systems for obtaining barley microspore-derived embryos, where embryos develop in liquid medium, on top of a solid medium layer or immobilized in a thin layer of agarose. While liquid culture systems allow the generation of large amounts of embryos for molecular analysis, solid culture systems are the ultimate tool for probing embryo development.

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References

  1. Wang M, van Bergen S, van Duijn B. Insights into a key developmental switch and its importance for efficient plant breeding. Plant Physiol 2000;124:523–530.

    Article  CAS  PubMed  Google Scholar 

  2. Hoekstra S, van Zijderveld MH, Louwerse JD, Heidekamp F, van der Mark F. Anther and microspore culture of Hordeum vulgare L. cv. Igri. Plant Sci 1992;86:89–96.

    Article  CAS  Google Scholar 

  3. Jähne-Gärtner A, Lörz H. Protocols for anther and microspore culture of barley. In: Hall RD, ed. Methods in Molecular Biology. Totowa, NJ: Humana, 1999:269–279.

    Google Scholar 

  4. Maraschin SF, Lamers GEM, de Pater BS, Spaink HP, Wang M. 14–3-3 isoforms and pattern formation during barley microspore embryogenesis. J Exp Bot 2003;51:1033–1043.

    Article  Google Scholar 

  5. Maraschin SF, Caspers M, Potokina E, Wülfert F, Graner A, Spaink HP, Wang M. cDNA array analysis of stress-induced gene expression in barley androgenesis. Physiol Plant 2006;127:535–550.

    Article  CAS  Google Scholar 

  6. Vrienten PL, Nakamura T, Kasha KJ. Characterization of cDNAs expressed in the early stages of microspore embryogenesis in barley (Hordeum vulgare) L. Plant Mol Biol 1999; 41:455–463.

    Article  Google Scholar 

  7. Maraschin SF, Vennik M, Lamers GEM, Spaink HP, Wang M. Time-lapse tracking of barley androgenesis reveals position-determined cell death within pro-embryos. Planta 2005;220:531–540.

    Article  CAS  Google Scholar 

  8. Kumlehn J, Lörz H. Monitoring sporophytic development of individual microspores of barley (Hordeum vulgare L.). In: Clement C, Pacini E, Audran JC, eds. Anther and Pollen: From Biology to Biotechnology. Berlin Heidelberg: Springer-Verlag, 1999:183–189.

    Google Scholar 

  9. Bolik M, Koop HU. Identification of embryogenic microspores of barley (Hordeum vulgare L.) by individual selection and culture and their potential for transformation by microinjection. Protoplasma 1991;162:61–68.

    Article  Google Scholar 

  10. Maraschin SF, Gaussand G, Pulido A, Olmedilla A, Lamers GEM, Korthout H, Spaink HP, Wang M. Programmed cell death during the transition from multicellular structures to globular embryos in barley androgenesis. Planta 2005;221: 459–470.

    Article  Google Scholar 

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© 2008 Humana Press, a part of Springer Science+Business Media, LLC

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Maraschin, S.F., van Bergen, S., Vennik, M., Wang, M. (2008). Culture and Time-Lapse Tracking of Barley Microspore-Derived Embryos. In: Suárez, M.F., Bozhkov, P.V. (eds) Plant Embryogenesis. Methods In Molecular Biology™, vol 427. Humana Press. https://doi.org/10.1007/978-1-59745-273-1_6

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  • DOI: https://doi.org/10.1007/978-1-59745-273-1_6

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-931-4

  • Online ISBN: 978-1-59745-273-1

  • eBook Packages: Springer Protocols

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