Immobilisation of barley aleurone layers enables parallelisation of assays and analysis of transient gene expression in single cells

Publikation: Forskning - peer reviewTidsskriftartikel

DOI

  • Kinga Zor
    Kinga ZorDanmarks Tekniske Universitet
  • Christina Mark
    Christina MarkDanmarks Tekniske Universitet
  • Arto Heiskanen
    Arto HeiskanenDanmarks Tekniske Universitet
  • Claus Krogh Madsen
  • Martin Dufva
    Martin DufvaDanmarks Tekniske Universitet
  • Jenny Emnéus
    Jenny EmnéusDanmarks Tekniske Universitet
  • Henrik Brinch-Pedersen
  • Christine Finnie
    Christine FinnieDanmarks Tekniske Universitet
The barley aleurone layer is an established model system for studying phytohormone signalling, enzyme secretion and programmed cell death during seed germination. Most analyses performed on the aleurone layer are end-point assays based on cell extracts, meaning each sample is only analysed at a single time point. By immobilising barley aleurone layer tissue on polydimethylsiloxane pillars in the lid of a multiwell plate, continuous monitoring of living tissue is enabled using multiple non-destructive assays in parallel. Cell viability and menadione reducing capacity were monitored in the same aleurone layer samples over time, in the presence or absence of plant hormones and other effectors. The system is also amenable to transient gene expression by particle bombardment, with simultaneous monitoring of cell death. In conclusion, the easy to handle and efficient experimental setup developed here enables continuous monitoring of tissue samples, parallelisation of assays and single cell analysis, with potential for time course studies using any plant tissue that can be immobilised, for example leaves or epidermal peels.
OriginalsprogEngelsk
TidsskriftPlant Physiology and Biochemistry
Vol/bind118
Sider (fra-til)71-76
Antal sider6
ISSN0981-9428
DOI
StatusUdgivet - sep. 2017

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