© 2007 The Author(s).
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RESEARCH PAPER |
Antisense-mediated suppression of C-hordein biosynthesis in the barley grain results in correlated changes in the transcriptome, protein profile, and amino acid composition
1Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, University of Aarhus, Research Centre Flakkebjerg, DK-4200 Slagelse, Denmark
2Center for Biological Sequence Analysis, BioCentrum, Technical University of Denmark, Building 208, DK-2800 Kgs. Lyngby, Denmark
* To whom correspondence should be addressed. E-mail: eva.vincze{at}agrsci.dk
Antisense- or RNAi-mediated suppression of the biosynthesis of nutritionally inferior storage proteins is a promising strategy for improving the amino acid profile of seeds. However, the potential pleiotropic effects of this on interconnected pathways and the agronomic quality traits need to be addressed. In the current study, a transcriptomic analysis of an antisense C-hordein line of barley was performed, using a grain-specific cDNA array. The C-hordein antisense line is characterized by marked changes in storage protein and amino acid profiles, while the seed weight is within the normal range and no external morphological irregularities were observed. The results of the transcriptome analysis showed excellent correlation with data on changes in the relative proportions of storage proteins and amino acid composition. The antisense line had a lower C-hordein level and down-regulated transcript encoding C-hordein. The production of the S-rich B/
- and D-hordeins was increased and significantly higher steady-state expression levels of the corresponding genes were observed. The increased synthesis of S-rich hordeins appeared to increase the demand for sulphur and the S-rich amino acids (cysteine and methionine), resulting in an up-regulation of key genes in the appropriate biosynthetic pathways. This study demonstrated the utility of the grain-specific cDNA microarray analysis to detect perturbations induced by antisense suppression of plant processes.
Key words: cDNA microarray, gene silencing, genetically modified (GM) crop, Hordeum vulgare, storage proteins, transcriptome
Received 9 August 2007; Accepted 19 September 2007
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