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

  • Simone de F. Maraschin
  • Sandra van Bergen
  • Marco Vennik
  • Mei Wang
Part of the Methods In Molecular Biology™ book series (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.

Keywords

Barley androgenesis microspore embryogenesis cell tracking liquid and solid culture 

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Copyright information

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

Authors and Affiliations

  • Simone de F. Maraschin
    • 1
  • Sandra van Bergen
    • 2
  • Marco Vennik
    • 2
  • Mei Wang
    • 1
  1. 1.Institute of BiologyLeiden University, Clusius LaboratoryThe Netherlands
  2. 2.Fytagoras BV and TNO Quality of LifeThe Netherlands

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