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Putrescine overproduction negatively impacts the oxidative state of poplar cells in culture
Authors:Sridev Mohapatra  Rakesh Minocha  Stephanie Long  Subhash C. Minocha
Affiliation:1. Department of Chemistry and Applied Biosciences of ETH Zurich, Center for Radiopharmaceutical Sciences ETH-PSI-USZ, Zurich, Switzerland;2. Department of Nuclear Medicine, University Hospital Zurich, Center for Radiopharmaceutical Sciences ETH-PSI-USZ, Zurich, Switzerland;3. University Hospital Zurich, Clinic for Cardiovascular Surgery, Zurich, Switzerland;4. University Hospital Zurich, Department of Nuclear Medicine, Cardiac Imaging, Zurich, Switzerland;5. Institute for Veterinary Pathology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland
Abstract:While polyamines (PAs) have been suggested to protect cells against Reactive Oxygen Species (ROS), their catabolism is known to generate ROS. We compared the activities of several enzymes and cellular metabolites involved in the ROS scavenging pathways in two isogenic cell lines of poplar (Populus nigra × maximowiczii) differing in their PA contents. Whereas the control cell line was transformed with β-glucuronidase (GUS), the other, called HP (High Putrescine), was transformed with a mouse ornithine decarboxylase (mODC) gene. The expression of mODC resulted in several-fold increased production of putrescine as well its enhanced catabolism. The two cell lines followed a similar trend of growth over the seven-day culture cycle, but the HP cells had elevated levels of soluble proteins. Accumulation of H2O2 was higher in the HP cells than the control cells, and so were the activities of glutathione reductase and monodehydroascorbate reductase; the activity of ascorbate peroxidase was lower in the former. The contents of reduced glutathione and glutamate were significantly lower in the HP cells but proline was higher on some days of analysis. There was a small difference in mitochondrial activity between the two cell lines, and the HP cells showed increased membrane damage. In the HP cells, increased accumulation of Ca was concomitant with lower accumulation of K. We conclude that, while increased putrescine accumulation may have a protective role against ROS in plants, enhanced turnover of putrescine actually can make them vulnerable to increased oxidative damage.
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