ABSTRACT
To investigate the involvement of Rhizobium etli cbb(3) oxidase in the response of Phaseolus vulgaris to drought, common bean plants were inoculated with the R. etli strain, CFNX713, overexpressing this oxidase in bacteroids (cbb(3)(+)) and subjected to drought conditions. The negative effect of drought on plant and nodule dryweight, nitrogen content, and nodule functionality was more pronounced in plants inoculated with the wild-type (WT) strain than in those inoculated with the cbb(3)(+) strain. Regardless of the plant treatment, bacteroids produced by the cbb(3)(+) strain showed higher respiratory capacity than those produced by the WT strain. Inoculation of plants with the cbb(3)(+) strain alleviated the negative effect of a moderate drought on the respiratory capacity of bacteroids and the energy charge of the nodules. Expression of the FixP and FixO components of the cbb(3) oxidase was higher in bacteroids of the cbb(3)(+) strain than in those of the WT strain under all experimental conditions. The decline in sucrose synthase activity and the decrease in dicarboxylic acids provoked by moderate drought stress were more pronounced in nodules from plants inoculated with the WT strain than in those inoculated with the cbb(3)(+) strain. Taken together, these results suggest that inoculation of plants with a R. etli strain having enhanced expression of cbb(3) oxidase in bacteroids reduces the sensitivity of P. vulgaris-R. etli symbiosis to drought and can modulate carbon metabolism in nodules.
Subject(s)
Electron Transport Complex IV/metabolism , Gene Expression Regulation, Bacterial , Nitrogen Fixation , Phaseolus/microbiology , Rhizobium etli/enzymology , Rhizobium etli/genetics , Adaptation, Physiological , Droughts , Phaseolus/growth & development , Phaseolus/physiology , SymbiosisABSTRACT
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.
Subject(s)
Medicago sativa/enzymology , Plant Leaves/enzymology , Plant Roots/enzymology , Superoxide Dismutase/genetics , Amino Acid Sequence , Antioxidants/metabolism , Base Sequence , Catalase/metabolism , Chloroplasts/metabolism , DNA, Complementary/chemistry , DNA, Complementary/genetics , Gene Expression Regulation, Enzymologic , Isoenzymes , Medicago sativa/genetics , Mitochondria/metabolism , Molecular Sequence Data , Peroxidases/metabolism , Phylogeny , Plant Leaves/genetics , Plant Roots/genetics , Plant Roots/microbiology , Plants, Genetically Modified , Protein Processing, Post-Translational , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sequence Analysis, DNA , Superoxide Dismutase/metabolism , Symbiosis , Transcription, GeneticABSTRACT
Sucrose synthase (SS) activity has been suggested to be a key point of regulation in nodule metabolism since this enzyme is down-regulated in response to different stresses which lead to decreased nitrogen fixation. In soybean, a dramatic decline of SS transcripts has been observed within 1 d from the onset of drought. Such a quick response suggests mediation by a signal transduction molecule. Abscisic acid (ABA) is a likely candidate to act as such a molecule as it mediates in a significant number of plant responses to environmental constraints. The hypothesis of ABA controlling nodule metabolism was approached in this work by assessing nodule responses to exogenous ABA supply in pea. Under the experimental conditions, ABA did not affect plant biomass, nodule numbers or dry weight. However, nitrogen fixation rate was reduced by 70% within 5 d and by 80% after 9 d leading to a reduced plant organic nitrogen content. Leghaemoglobin (Lb) content declined in parallel with that of nitrogen fixation. SS activity, however, was not affected by ABA treatment, and neither were the activities of the enzymes aspartate amino transferase, alkaline invertase, malate dehydrogenase, glutamate synthase, uridine diphosphoglucose pyrophosphorylase, isocitrate dehydrogenase, and glutamine synthetase. Nodule bacteroid-soluble protein content was reduced in nodules only after 9 d of ABA treatment. These results do not support the hypothesis that ABA directly regulates SS activity. However, they do suggest the occurrence of at least two different control pathways in nodules under environmental constraints, which include ABA being involved in a Lb/oxygen-related control of nitrogen fixation.