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Journal of Experimental Botany 2009 60(4):1333-1347; doi:10.1093/jxb/erp023
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© 2009 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

RESEARCH PAPER

A roadmap for zinc trafficking in the developing barley grain based on laser capture microdissection and gene expression profiling

Birgitte Tauris, Søren Borg*, Per L. Gregersen and Preben B. Holm

University of Aarhus, Facultry of Agricultural Sciences, Department of Genetics and Biotechnology, Forsøgsvej 1, 4200 Slagelse, Denmark

* To whom correspondence should be addressed: soren.borg{at}agrsci.dk

Nutrients destined for the developing cereal grain encounter several restricting barriers on their path towards their final storage sites in the grain. In order to identify transporters and chelating agents that may be involved in transport and deposition of zinc in the barley grain, expression profiles have been generated of four different tissue types: the transfer cells, the aleurone layer, the endosperm, and the embryo. Cells from these tissues were isolated with the ‘laser capture microdissection’ technology and the extracted RNA was subjected to three rounds of T7-based amplification. The amplified RNA was subsequently hybridized to Affymetrix 22K Barley GeneChips. Due to the short average length of the amplified transcripts and the positioning of numerous probe sets at locations more than 400 base pairs (bp) from the poly(A)-tail, a normalization approach was used where the probe positions were taken into account. On the basis of the expression levels of a number of metal homeostasis genes, a working model is proposed for the translocation of zinc from the phloem to the storage sites in the developing grain.

Key words: Affymetrix, gene expression profiling, laser capture microdissection, microarray, RNA amplification, zinc homeostasis

Received 21 November 2008; Accepted 7 January 2009


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