Use of cell morphology to evaluate the effect of a peroxidase gene on cell death induction thresholds in tobacco

Abstract

Tobacco suspension cultures were subjected to a range of heat stresses and used to compare morphological aspects of programmed cell death (PCD) and necrosis. Cells undergoing PCD were found to display characteristic death morphology, caused by cytoplasmic retraction of the protoplast, and to have cleaved DNA. We evaluated if the morphological characteristics of PCD could be used to monitor changes in cell death induction thresholds in transgenic cell cultures with high levels of peroxidase activity. Again, using a heat shock assay, we show that tobacco cell cultures with elevated levels of peroxidase have higher cell death induction threshold levels than wild type tobacco cell cultures. Thus, assessing PCD associated morphological changes can report on the effect of altering peroxidase genes on cell death activation in tobacco. This study demonstrates that PCD morphology could routinely be used to monitor the effects of introduced genes on programmed cell death induction thresholds in plants

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