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Expression Studies of Superoxide Dismutases in Nodules and Leaves of Transgenic Alfalfa Reveal Abundance of Iron-Containing Isozymes, Posttranslational Regulation, and Compensation of Isozyme Activities

October 2001 , Volume 14 , Number  10
Pages  1,178 - 1,188

Maria C. Rubio , 1 Javier Ramos , 1 K. Judith Webb , 2 Frank R. Minchin , 2 Esther González , 3 Cesar Arrese-Igor , 3 and Manuel Becana 1

1Departamento de Nutrición Vegetal, Estación Experimental de Aula Dei, Consejo Superior de Investigaciones Científicas, Apdo. 202, 50080 Zaragoza, Spain; 2Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth SY23 3EB, U.K.; 3Departamento de Ciencias del Medio Natural, Universidad Pública de Navarra, 31006 Pamplona, Spain

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Accepted 28 June 2001.

The composition of antioxidant enzymes, especially superoxide dismutase (SOD), was studied in one nontransgenic and three transgenic lines of nodulated alfalfa plants. Transgenic lines overproduced MnSOD in the mitochondria of nodules and leaves (line 1--10), MnSOD in the chloroplasts (line 4--6), and FeSOD in the chloroplasts (line 10--7). In nodules of line 10--7, the absence of transgene-encoded FeSOD activity was due to a lack of mRNA, whereas in nodules of line 4-6 the absence of transgene-encoded MnSOD activity was due to enzyme inactivation or degradation. Transgenic alfalfa showed a novel compensatory effect in the activities of MnSOD (mitochondrial) and FeSOD (plastidic) in the leaves, which was not caused by changes in the mRNA levels. These findings imply that SOD activity in plant tissues and organelles is regulated, at least partially, at the posttranslational level. All four lines had low CuZnSOD activities and an abundant FeSOD isozyme, especially in nodules, indicating that FeSOD performs important antioxidant functions other than the scavenging of superoxide radicals generated in photosynthesis. This was confirmed by the detection of FeSOD cDNAs and proteins in nodules of other legumes such as cowpea, pea, and soybean. The cDNA encoding alfalfa nodule FeSOD was characterized and the deduced protein found to contain a plastid transit peptide. A comparison of sequences and other properties reveals that there are two types of FeSODs in nodules.

© 2001 The American Phytopathological Society