Microarray analysis of gene expression in germinating barley embryos (Hordeum vulgare L.).

Funct Integr Genomics

Molecular Plant Breeding CRC, Centre for Plant Conservation Genetics, Southern Cross University, Lismore, NSW, Australia.

Published: July 2005

AI Article Synopsis

  • A cDNA library with about 5,000 clones from germinating barley embryos was created to study gene expression changes during the first 4 days of seed soaking.
  • Microarray analysis compared gene expression profiles from different time points postimbibition, revealing that while most genes were stable at early stages, significant fluctuations occurred between 24 to 96 hours.
  • The findings support theories on genetic changes necessary for embryo development and confirm previous studies regarding the stress-response protein JIP-23, linking mRNA levels to protein quantities.

Article Abstract

A cDNA library containing approximately 5,000 clones from germinating barley embryos was constructed and used to examine the variation in gene expression patterns during the first 4 days postimbibition. The expression profiles of embryos (including scutellum) from 4 to 96 h postimbibition were compared to a reference profile from 24 h postimbibition using microarray analysis. A subset of clones exhibiting tenfold or greater differential expression patterns was sequenced to elucidate function. All of the sequenced clones could be identified to at least EST level with 64% exhibiting homology to published protein sequences. Almost 95% of the library exhibited similar expression levels at the 4 h time point as at the 24 h reference point. From 24 to 96 h, however, considerable fluctuations in gene expression occurred. The observed patterns of gene expression for the classified genes are consistent with the expected genetic changes required to prepare an embryo for germinative development. A replicate set of clones for the 23-kDa jasmonate-induced protein was identified. The current data not only provides conclusive evidence for the expression patterns of this abundant stress-response protein in germinating embryos, but also serves to validate previous research into JIP-23 isoforms, function and the relationship between timing of mRNA upregulation and protein abundance.

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http://dx.doi.org/10.1007/s10142-005-0133-6DOI Listing

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