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PLANT PHYSIOLOGY , Vol 113, Issue 4 1177-1183, Copyright © 1997 by American Society of Plant Biologists
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WHOLE PLANT, ENVIRONMENTAL, AND STRESS PHYSIOLOGY |
Increased Resistance to Oxidative Stress in Transgenic Plants by Targeting Mannitol Biosynthesis to Chloroplasts
B. Shen, R. G. Jensen and H. J. Bohnert
Department of Plant Sciences (B.S., R.G.J., H.J.B.), Department of Biochemistry (R.G.J., H.J.B.), and Department of Molecular and Cellular Biology (H.J.B.), The University of Arizona, Tucson, Arizona 85721
To investigate the potential role of a polyol, mannitol, in oxidative
stress protection, a bacterial mannitol-1-phosphate dehydrogenase gene was
targeted to chloroplasts by the addition of an amino-terminal transit
peptide. Transgenic tobacco (Nicotiana tabacum) lines accumulate mannitol
at concentrations ranging from 2.5 to 7 [mu]mol/g fresh weight. Line BS1-31
accumulated approximately 100 mM mannitol in chloroplasts and was identical
to the wild type in phenotype and photosynthetic performance. The presence
of mannitol in chloroplasts resulted in an increased resistance to methyl
viologen (MV)-induced oxidative stress, documented by the increased
retention of chlorophyll in transgenic leaf tissue following MV treatment.
In the presence of MV, isolated mesophyll cells of BS1-31 exhibited higher
CO2 fixation than the wild type. When the hydroxyl radical probe dimethyl
sulfoxide was introduced into cells, the initial formation rate of methane
sulfinic acid was significantly lower in cells containing mannitol in the
chloroplast compartment than in wild-type cells, indicating an increased
hydroxyl radical-scavenging capacity in BS1-31 tobacco. We suggest that the
chloroplast location of mannitol can supplement endogenous
radical-scavenging mechanisms and reduce oxidative damage of cells by
hydroxyl radicals.
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