Abstract
In this work, we expressed an Arabidopsis thaliana-coded protein (AKR4C9) in transgenic barley to study its enzymatic activity and to enhance the reactive aldehyde neutralizing capacity (part of the oxidative stress tolerance) of transgenic plants. Total leaf protein was extracted from transgenic plants expressing either C or N-terminally His-tagged aldo–keto reductase (AKR) enzyme and purified by affinity chromatography. The Arabidopsis-coded enzyme showed moderate activity against the synthetic reactive aldehyde, glutaraldehyde, and low but detectable enzyme activity against fructose with a low Michaelis–Menten constant (Km value). Activity of the C and the N-terminally His-tagged AKRs were found to be in the same range. Glutaraldehyde was also tested in vivo by spraying onto the leaves of the plants. The reactive aldehyde tolerance of both wild type and transgenic plants, as well as the general physiological effects of this reactive aldehyde treatment were evaluated. The growth rate was found to decrease in all (both wild type and transgenic) plants. The high AKR-expressing transgenic plants showed a lower respiratory rate, and they also showed higher fresh weight, higher chlorophyll content and photosynthetic activity, indicating a higher reactive aldehyde tolerance. Cadmium (Cd) treatment was also performed to validate this result. Cd caused strong lipid peroxidation; however, the Arabidopsis enzyme lowered the reactive aldehyde content as expected. This is the first report in which kinetic parameters of the fructose reduction by the stress inducible plant AKR enzyme are presented. Furthermore, data on the effects of a reactive aldehyde treatment on intact plants are also provided.
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Acknowledgments
The authors are grateful to Gábor V. Horváth (Biological Research Centre, Szeged Hungary) for his scientific advice and for providing the vector-cloned At2g37770.2 gene. We would like to thank Wendy Harwood (JIC, Norwich, UK) for the barley transformation method. We also thank Ming-Bo Wang (CSIRO Plant Industry, Canberra) for the pWBVec8 vector. We acknowledge the valuable help of Ádám Solti (Department of Plant Physiology, ELTE) discussing the results gained on the whole plant. The project was supported by the European Union and co-financed by the European Social Fund (Grant Agreement No. TAMOP 4.2.1/B-09/1/KMR-2010-0003). László Tamás is in receipt of grants from the Hungarian Scientific Fund (OTKA T-46703 and K-67844).
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Éva, C., Tóth, G., Oszvald, M. et al. Overproduction of an Arabidopsis aldo–keto reductase increases barley tolerance to oxidative and cadmium stress by an in vivo reactive aldehyde detoxification. Plant Growth Regul 74, 55–63 (2014). https://doi.org/10.1007/s10725-014-9896-x
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DOI: https://doi.org/10.1007/s10725-014-9896-x