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JXB Advance Access originally published online on September 12, 2006
Journal of Experimental Botany 2006 57(14):3601-3618; doi:10.1093/jxb/erl111
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Published by Oxford University Press [2006] on behalf of the Society for Experimental Biology

RESEARCH PAPER

Additional freeze hardiness in wheat acquired by exposure to –3 °C is associated with extensive physiological, morphological, and molecular changes

Eliot M. Herman1,2,*, Kelsi Rotter1, Ramaswamy Premakumar3, G Elwinger3, Rino Bae2, Linda Ehler-King2, Sixue Chen4 and David P. Livingston, III3

1Plant Genetics Research Unit, USDA/ARS, Donald Danforth Plant Science Center, 975 N. Warson Rd, St Louis, MO 63132, USA
2Climate Stress Laboratory, USDA/ARS, Beltsville, MD 20705, USA
3USDA/ARS, and North Carolina State University, Raleigh, NC 27695, USA
4Proteomics Facility, Donald Danforth Plant Science Center, 975 N. Warson Rd, St Louis, MO 63132, USA

* To whom correspondence should be addressed. E-mail: eherman{at}danforthcenter.org

Cold-acclimated plants acquire an additional 3–5 °C increase in freezing tolerance when exposed to –3 °C for 12–18 h before a freezing test (LT50) is applied. The –3 °C treatment replicates soil freezing that can occur in the days or weeks leading to overwintering by freezing-tolerant plants. This additional freezing tolerance is called subzero acclimation (SZA) to differentiate it from cold acclimation (CA) that is acquired at above-freezing temperatures. Using wheat as a model, results have been obtained indicating that SZA is accompanied by changes in physiology, cellular structure, the transcriptome, and the proteome. Using a variety of assays, including DNA arrays, reverse transcription–polymerase chain reaction (RT–PCR), 2D gels with mass spectroscopic identification of proteins, and electron microscopy, changes were observed to occur as a consequence of SZA and the acquisition of added freezing tolerance. In contrast to CA, SZA induced the movement of intracellular water to the extracellular space. Many unknown and stress-related genes were upregulated by SZA including some with obvious roles in SZA. Many genes related to photosynthesis and plastids were downregulated. Changes resulting from SZA often appeared to be a loss of rather than an appearance of new proteins. From a cytological perspective, SZA resulted in alterations of organelle structure including the Golgi. The results indicate that the enhanced freezing tolerance of SZA is correlated with a wide diversity of changes, indicating that the additional freezing tolerance is the result of complex biological processes.

Key words: Aquaporin, cold acclimation, DNA array, electron microscopy, freeze hardiness, proteome, proteomics, transcriptome, wheat


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