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JXB Advance Access published online on February 8, 2006

Journal of Experimental Botany, doi:10.1093/jxb/erj072
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© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. The online version of this article has been published under an Open Access model. Users are entitled to use, reproduce, disseminate, or display the Open Access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and the Society for Experimental Biology are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact: journals.permissions@oxfordjournals.org
Received September 14, 2005
Accepted November 16, 2005

RESEARCH PAPER

{alpha}-Amylase and programmed cell death in aleurone of ripening wheat grains

Kolumbina Mrva 1, Meredith Wallwork 2, and Daryl J. Mares 1 *

1 Plant and Pest Science, School of Agriculture, Food and Wine, University of Adelaide, Waite Campus, Glen Osmond, SA 5064, Australia
2 Adelaide Microscopy, University of Adelaide, Medical School North, Frome Road, Adelaide, SA 5005, Australia

* To whom correspondence should be addressed.
Daryl J. Mares, E-mail: daryl.mares{at}adelaide.edu.au


   Abstract

Late maturity {alpha}-amylase (LMA) in wheat is a genetic defect that may result in the accumulation of unacceptable levels of high pI {alpha}-amylase in grain in the absence of germination or weather damage. During germination, gibberellin produced in the embryo triggers expression of {alpha}-Amy genes, the synthesis of {alpha}-amylase and, subsequently, cell death in the aleurone. LMA also involves the aleurone and whilst LMA appears to be independent of the embryo there is nevertheless some evidence that gibberellin is involved. The aim of this investigation was to determine whether the increase in {alpha}-amylase activity in LMA-prone genotypes, like {alpha}-amylase synthesis by aleurone cells in germinating or GA-challenged grains, is followed by aleurone cell death. Programmed cell death was seen in aleurone layers from developing, ripe and germinated grains using confocal microscopy and fluorescent probes specific for dead or living cells. Small pockets of dying cells were observed distributed at random throughout the aleurone of ripening LMA-affected grains and by harvest-ripeness these cells were clearly dead. The first appearance of dying cells, 35 d post-anthesis, coincided with the later part of the ‘window of sensitivity’ in grain development in LMA-prone wheat cultivars. No dead or dying cells were present in ripening or fully ripe grains of control cultivars. In germinating grains, dying cells were observed in the aleurone adjacent to the scutellum and, as germination progressed, the number of dead cells increased and the affected area extended further towards the distal end of the grain. Aside from the obvious differences in spatial distribution, dying cells in 20-24 h germinated grains were similar to dying cells in developing LMA-affected grains, consistent with previous measurements of {alpha}-amylase activity. The increase in high pI {alpha}-amylase activity in developing grains of LMA-prone cultivars, like {alpha}-amylase synthesis in germinating grains, is associated with cell death, providing further evidence for the involvement of gibberellin in the LMA response.

Keywords: Aleurone layer; cell viability; germination; late maturity {alpha}-amylase; wheat.
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