title: Sensitive whole mount in situ localization of small RNAs in plants.
abstract: Small RNAs, such as microRNAs (miRNAs), regulate gene expression and play important roles in many plant processes. Although our knowledge of their biogenesis and mode of action has significantly progressed, we still have comparatively little information about their biological functions. In particular, knowledge about their spatio-temporal expression patterns rely on either indirect detection by use of reporter constructs or labor-intensive direct detection by in situ hybridization on sectioned material. None of the current approaches allows a systematic investigation of small RNA expression patterns. Here, we present a sensitive method for in situ detection of miRNAs and siRNAs in intact plant tissues that utilizes both double-labeled probes and a specific cross-linker. We determined the expression patterns of several small RNAs in diverse plant tissues.
authors.affiliation: Center for Organismal Studies (COS), University of Heidelberg, Im Neuenheimer Feld 230, 69120, Heidelberg, Germany.
authors.forename: Mouli
authors.initials: M
authors.lastname: Ghosh Dastidar
authors.affiliation: Gregor Mendel Institute (GMI), Austrian Academy of Sciences, Vienna Biocenter (VBC), Dr. Bohr-Gasse 3, 1030, Vienna, Austria.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Mosiolek
authors.affiliation: Institute for Cell Biology and Plant Biochemistry, University of Regensburg, Universitaetsstrasse 31, 93053, Regensburg, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Bleckmann
authors.affiliation: Institute for Cell Biology and Plant Biochemistry, University of Regensburg, Universitaetsstrasse 31, 93053, Regensburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Dresselhaus
authors.affiliation: Gregor Mendel Institute (GMI), Austrian Academy of Sciences, Vienna Biocenter (VBC), Dr. Bohr-Gasse 3, 1030, Vienna, Austria.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Nodine
authors.affiliation: Center for Organismal Studies (COS), University of Heidelberg, Im Neuenheimer Feld 230, 69120, Heidelberg, Germany.
authors.forename: Alexis
authors.identifier: 0000-0001-6843-1059
authors.initials: A
authors.lastname: Maizel
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D017403:In Situ Hybridization; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; D035683:MicroRNAs; D018749:RNA, Plant; D034741:RNA, Small Interfering
keywords: 
  Arabidopsis thaliana
  ; N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride; expression pattern; in situ hybridization; microRNA; short interfering RNA; technical advance