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Published on September 10, 2008; 10.1104/pp.108.127001

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Received August 4, 2008
Accepted September 5, 2008

Different hormonal regulation of cellular differentiation and function in nucellar projection and endosperm transfer cells – a microdissection-based transcriptome study of young barley grains

Johannes Thiel *, Diana Weier , Nese Sreenivasulu , Marc Strickert , Nicola Weichert , Michael Melzer , Tobias Czauderna , Ulrich Wobus , Hans Weber , and Winfriede Weschke

Leibniz-Institut fur Pflanzengenetik und Kulturpflanzenforschung (IPK), Corrensstraße 3, D-06466 Gatersleben, Germany

* Corresponding author; email: thielj{at}ipk-gatersleben.de.

Nucellar projection (NP) and endosperm transfer cells (ETC) are essential tissues in growing barley grains responsible for nutrient transfer from maternal to filial tissues, endosperm /embryo nutrition and grain development. A laser microdissection pressure catapulting (LMPC)-based transcriptome analysis was established to study NP and ETC separately using a barley 12k macroarray. A major challenge was to isolate high-quality mRNA from pre-embedded, fixed tissue, while maintaining tissue integrity. We show that probes generated from fixed and embedded tissue sections represent largely the transcriptome (>70%) of non-chemically treated and non-amplified references. In NP, the top-down gradient of cellular differentiation is reflected by expression of C3HC4-type ubiquitin ligases and different histone genes, cell-wall biosynthesis and expansin/extensin genes as well as genes involved in programmed cell death-related proteolysis coupled to N-remobilization, indicating distinct areas simultaneously undergoing mitosis, cell elongation and disintegration. Activated gene expression related to gibberellin synthesis and function suggests a regulatory role for gibberellins (GAs) in establishment of the differentiation gradient. Upregulation of plasmalemma-intrinsic protein- and tonoplast-intrinsic protein-genes indicates involvement in nutrient transfer and/or unloading. In ETC, AP2/EREBP-like transcription factors and ethylene functions are transcriptionally activated, a response possibly coupled to activated defence mechanisms. Transcriptional activation of nucleotide sugar metabolism may be attributed to ascorbate synthesis and/or cell-wall biosynthesis. These processes are potentially controlled by trehalose-6-phosphate synthase/phosphatase as suggested by expression of respective genes. Upregulation of amino acid permeases in endosperm transfer cells indicates important roles in active nutrient uptake from the apoplastic space into the endosperm.




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