Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/107078
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dc.contributor.authorBetts, N.en
dc.contributor.authorBerkowitz, O.en
dc.contributor.authorLiu, R.en
dc.contributor.authorCollins, H.en
dc.contributor.authorSkadhauge, B.en
dc.contributor.authorDockter, C.en
dc.contributor.authorBurton, R.en
dc.contributor.authorWhelan, J.en
dc.contributor.authorFincher, G.en
dc.date.issued2017en
dc.identifier.citationThe Plant Journal, 2017; 91(4):754-765en
dc.identifier.issn0960-7412en
dc.identifier.issn1365-313Xen
dc.identifier.urihttp://hdl.handle.net/2440/107078-
dc.descriptionPublished online 16 May 2017en
dc.description.abstractIsolated barley (Hordeum vulgare L.) aleurone layers have been widely used as a model system for studying gene expression and hormonal regulation in germinating cereal grains. A serious technological limitation of this approach has been the inability to confidently extrapolate conclusions obtained from isolated tissues back to the whole grain, where the co-location of several living and non-living tissues results in complex tissue-tissue interactions and regulatory pathways coordinated across the multiple tissues. Here we have developed methods for isolating fragments of aleurone, starchy endosperm, embryo, scutellum, pericarp-testa, husk and crushed cell layers from germinated grain. An important step in the procedure involves the rapid fixation of the intact grain to freeze the transcriptional activity of individual tissues while dissection is effected for subsequent transcriptomic analyses. The developmental profiles of 19 611 gene transcripts were precisely defined in the purified tissues and in whole grain during the first 24 h of germination by RNA sequencing. Spatial and temporal patterns of transcription were validated against well-defined data on enzyme activities in both whole grain and isolated tissues. Transcript profiles of genes involved in mitochondrial assembly and function were used to validate the very early stages of germination, while the profiles of genes involved in starch and cell wall mobilisation matched existing data on activities of corresponding enzymes. The data will be broadly applicable for the interrogation of co-expression and differential expression patterns and for the identification of transcription factors that are important in the early stages of grain and seed germination.en
dc.description.statementofresponsibilityNatalie S. Betts, Oliver Berkowitz, Ruijie Liu, Helen M. Collins, Birgitte Skadhauge, Christoph Dockter, Rachel A. Burton, James Whelan and Geoffrey B. Fincheren
dc.language.isoenen
dc.publisherWiley-Blackwellen
dc.rights© 2017 The Authors. The Plant Journal © 2017 John Wiley & Sons Ltden
dc.subjectbarley; Hordeum vulgare; germination; dissection; mitochondria; RNA-seq; aleurone; scutellum; technical advanceen
dc.titleIsolation of tissues and preservation of RNA from intact, germinated barley grainen
dc.typeJournal articleen
dc.identifier.rmid0030069971en
dc.identifier.doi10.1111/tpj.13600en
dc.relation.granthttp://purl.org/au-research/grants/arc/LP130100600en
dc.identifier.pubid352748-
pubs.library.collectionAgriculture, Food and Wine publicationsen
pubs.library.teamDS03en
pubs.verification-statusVerifieden
pubs.publication-statusPublisheden
dc.identifier.orcidBetts, N. [0000-0002-3345-8160]en
dc.identifier.orcidCollins, H. [0000-0003-3885-7707]en
dc.identifier.orcidBurton, R. [0000-0002-0638-4709]en
Appears in Collections:Agriculture, Food and Wine publications

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