Plant Embryogenesis

Volume 427 of the series Methods In Molecular Biology™ pp 77-89

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

  • Simone de F. MaraschinAffiliated withInstitute of Biology, Leiden University, Clusius Laboratory
  • , Sandra van BergenAffiliated withFytagoras BV and TNO Quality of Life
  • , Marco VennikAffiliated withFytagoras BV and TNO Quality of Life
  • , Mei WangAffiliated withInstitute of Biology, Leiden University, Clusius Laboratory

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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.


Barley androgenesis microspore embryogenesis cell tracking liquid and solid culture