In order to better understand the role of antioxidant enzymes in plant stress protection mechanisms, transgenic tobacco ( Nicotiana tabacum cv. Xanthi) plants were developed that overexpress both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts. These plants were evaluated for protection against methyl viologen (MV, paraquat)-mediated oxidative damage both in leaf discs and whole plants. Transgenic plants that express either chloroplast-targeted CuZnSOD (C) or MnSOD (M) and APX (A) were developed (referred to as CA plants and AM plants, respectively). These plant lines were crossed to produce plants that express all three transgenes (CMA plants and AMC plants). These plants had higher total APX and SOD activities than non-transgenic (NT) plants and exhibit novel APX and SOD isoenzymes not detected in NT plants. As expected, transgenic plants that expressed single SODs showed levels of protection from MV that were only slightly improved compared to NT plants. The expression of either SOD isoform along with APX led to increased protection while expression of both SODs and APX provided the highest levels of protection against membrane damage in leaf discs and visual symptoms in whole plants.

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Six peroxidase (POD) cDNAs were isolated from suspension cultures of sweetpotato (Ipomoea batatas) by cDNA library screening, and their expression was investigated with a view to understanding the physiological functions of each POD in relation to environmental stress. The gene products encoded by these cDNAs could be divided into two groups, anionic PODs (SWPA4, SWPA5, SWPA6) and basic PODs (SWPB1, SWPB2, SWPB3), on the basis of the predicted pI values of the mature proteins. RT-PCR analysis revealed that the six POD genes showed diverse expression patterns in various tissues of intact plants, a various stages of growth in suspension cultures, and in leaf tissues exposed to different stresses. The six genes from which they were derived are predominantly expressed in cultured cells of sweetpotato. Thus, transcripts of swpa4 were not detected in any tissues of the intact plant. The genes swpa6 and swpb1 were highly expressed in root tissues, whereas swpa6 and swpb3 were highly expressed in stem tissues. During suspension culture, the expression patterns of the six genes differed from each other. The level of swpa4, swpa5, swpb2 and swpb3 transcripts progressively increased during culture, whereas swpa6 and swpb1 showed high expression levels regardless of the age of the culture. In leaf tissues the six POD genes responded differently to various abiotic stresses. In particular, swpa4 was highly induced by several abiotic stresses, including exposure to hydrogen peroxide (440 mM) or NaCl (100 mM), and wounding of leaf tissues, suggesting that this POD gene is inducible by many stresses. Based on the different expression patterns of these POD genes, we propose that each POD may have different enzymatic properties and physiological functions during cell growth and development.