title: Different hormonal regulation of cellular differentiation and function in nucellar projection and endosperm transfer cells: a microdissection-based transcriptome study of young barley grains.
abstract: Nucellar projection (NP) and endosperm transfer cells (ETC) are essential tissues in growing barley (Hordeum vulgare) grains, responsible for nutrient transfer from maternal to filial tissues, endosperm/embryo nutrition, and grain development. A laser microdissection pressure catapulting-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 preembedded, fixed tissue while maintaining tissue integrity. We show that probes generated from fixed and embedded tissue sections represent largely the transcriptome (>70%) of nonchemically treated and nonamplified references. In NP, the top-down gradient of cellular differentiation is reflected by the 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 nitrogen 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 in establishment of the differentiation gradient. Up-regulation 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 defense 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-P synthase/phosphatase, as suggested by expression of their respective genes. Up-regulation of amino acid permeases in ETC indicates important roles in active nutrient uptake from the apoplastic space into the endosperm.
authors.affiliation: Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung, D-06466 Gatersleben, Germany. thielj@ipk-gatersleben.de
authors.forename: Johannes
authors.initials: J
authors.lastname: Thiel
authors.forename: Diana
authors.initials: D
authors.lastname: Weier
authors.forename: Nese
authors.initials: N
authors.lastname: Sreenivasulu
authors.forename: Marc
authors.initials: M
authors.lastname: Strickert
authors.forename: Nicola
authors.initials: N
authors.lastname: Weichert
authors.forename: Michael
authors.initials: M
authors.lastname: Melzer
authors.forename: Tobias
authors.initials: T
authors.lastname: Czauderna
authors.forename: Ulrich
authors.initials: U
authors.lastname: Wobus
authors.forename: Hans
authors.initials: H
authors.lastname: Weber
authors.forename: Winfriede
authors.initials: W
authors.lastname: Weschke
mesh_terms: D002454:Cell Differentiation; D017343:Genes, Plant; D001467:Hordeum / Q000166:cytology* / Q000235:genetics* / Q000502:physiology; D010937:Plant Growth Regulators / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics*
chemical_list: D010937:Plant Growth Regulators; D012333:RNA, Messenger
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