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HvPG1 and ECA1: two genes activated transcriptionally in the transition of barley microspores from the gametophytic to the embryogenic pathway

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Abstract

Microspores genetically programmed to produce male gametes can be switched to the embryogenic pathway to give rise to haploid embryos. Microspore embryogenesis is usually induced in barley by stress pre-treatment applied to vacuolated microspores. We studied the expression of two genes during the early stages of microspore embryogenesis to gain further insight into the microspore transition from the gametophytic to the embryogenic pathway. RT-PCR together with in situ hybridization on sections (ISH) and whole-mount in situ hybridization (WISH) were used to analyse the expression of the early-culture abundant gene (ECA1), which is expressed in barley during microspore embryogenesis, and a polygalacturonase gene (HvPG1), a late pollen gene expressed during gametogenesis only after microspore division. Both ECA1 and HvPG1 genes were transcriptionally active after stress pre-treatment in the same populations of microspore-derived structures, representing the sporophytically induced ones. ECA1 transcripts were also detected after 3 days’ culture. Our results point to the possibility of using ECA1 gene expression as a marker for the induction of microspore embryogenesis and the earliest stages of this process. Finally, we demonstrate that WISH is a suitable technique for studying gene expression in embryogenic microspore populations and, because different structures can be examined individually, is an appropriate complement to transcriptomic profile analyses in the study of early microspore embryogenesis.

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Acknowledgments

This study was supported by a Spain–Hungary co-operation project REF2006HU0004, the Spanish MEC project (Ref. BFU2006-09876/BFI) and by the grant “Elnöki Pályázat 2006” from the Hungarian Academy of Sciences. A. Pulido received a fellowship from the Junta de Andalucía, Spain. We thank Dr. Yves Henry for his critical review of our paper and Dr. J. Trout for revising our English text.

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Correspondence to Adela Olmedilla.

Additional information

Communicated by A. Feher.

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Pulido, A., Bakos, F., Devic, M. et al. HvPG1 and ECA1: two genes activated transcriptionally in the transition of barley microspores from the gametophytic to the embryogenic pathway. Plant Cell Rep 28, 551–559 (2009). https://doi.org/10.1007/s00299-008-0662-2

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Keywords

  • Barley
  • Gametophytic development
  • Molecular marker
  • Microspore embryogenesis
  • Whole-mount in situ hybridization