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JXB Advance Access published online on February 13, 2004
Journal of Experimental Botany, doi:10.1093/jxb/erh074
© 2004 by Oxford University Press
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Received July 17, 2003; accepted November 25, 2003
© 2004 Society for Experimental Biology

Research paper

The effect of reduced glutathione on morphology and gene expression of white spruce (Picea glauca) somatic embryos

Claudio Stasolla 1*, Mark F. Belmonte 2, Leonel van Zyl 3, Deborah L. Craig 3, Wenbin Liu 4, Edward C. Yeung 2, and Ronald R. Sederoff 3

1 Department of Plant Science, University of Manitoba, Winnipeg, R3T 2N2 Manitoba, Canada
2 Department of Biological Sciences, University of Calgary, Calgary, T2N 1N4 Alberta, Canada
3 Forest Biotechnology Group, Department of Forestry, North Carolina State University, Raleigh, NC 27695-7247, USA
4 Baylor College of Medicine, 452A One Baylor Plaza, Houston, TX 77030, USA

* To whom correspondence should be addressed. E-mail: stasolla{at}ms.umanitoba.ca.


  Abstract

Inclusions of reduced glutathione (GSH) in the maturation medium increased the conversion frequency of white spruce somatic embryos without the need of a partial drying treatment (PDT). This beneficial effect was the result of major alterations in morphology and gene expression during the maturation period. Compared with control embryos, GSH-treated embryos showed a differential accumulation of storage products, i.e. preferential deposition of starch, the reduced formation of protein bodies, and increased vacuolation of cells. These morphological changes correlated with extensive alterations of gene expression occurring throughout the maturation period. The transcript profiles of stage-specific embryos matured with or without GSH were analysed using a DNA microarray containing 2 178 cDNAs from loblolly pine (Pinus taeda). The efficiency of heterologous hybridization between spruce and pine species on microarrays has previously been documented. The results indicate that several genes involved in a variety of signal regulatory pathways were differentially expressed in developing GSH- treated embryos. The transcript levels of many genes involved in carbohydrate metabolism and protein synthesis were altered by the presence of GSH and denoted differences in physiology between treatments. Extensive changes in the expression of genes participating in hormone synthesis, nucleotide metabolism, and meristem formation were also observed and related to the post-embryonic performance of the embryos.

Key words: Embryogenesis, glutathione, microarray, transcript levels.


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