A total of 17 culturable nitrogen-fixing bacterial strains associated with the roots of wheat growing in different regions of Greece were isolated and characterized for plant-growth-promoting traits such as auxin production and phosphate solubilization. The phylogenetic position of the isolates was first assessed by the analysis of the PCR-amplified 16S rRNA gene. The comparative sequence analysis and phylogenetic analysis based on 16S rRNA gene sequences show that the isolates recovered in this study are grouped with Azospirillum brasilense, Azospirillum zeae, and Pseudomonas stutzeri. The diazotrophic nature of all isolates was confirmed by amplification of partial nifH gene sequences. The phylogenetic tree based on nifH gene sequences is consistent with 16S rRNA gene phylogeny. The isolates belonging to Azospirillum species were further characterized by examining the partial dnaK gene phylogenetic tree. Furthermore, it was demonstrated that the ipdC gene was present in all Azospirillum isolates, suggesting that auxin is mainly synthesized via the indole-3-pyruvate pathway. Although members of P. stutzeri and A. zeae are known diazotrophic bacteria, to the best of our knowledge, this is the first report of isolation and characterization of strains belonging to these bacterial genera associated with wheat.
The utilization of some agro-industrial wastes as soil conditioners to provide free-living nitrogen-fixing bacterial populations (e.g. Azospirillum spp.) with carbon and energy sources, may be an interesting perspective for agriculture. However, the presence of ammonium nitrogen in cultivated soils and/or various wastes could inhibit the growth of the nitrogen-fixing populations. The present investigation shows that growth of Azospirillum lipoferum was restricted at a dissolved oxygen (DO) concentration equal to 135 microM, when the initial NH4Cl concentration increased from 0.5 to 0.9 g/l. The activities of both citrate synthase (CS) and isocitrate dehydrogenase were significantly decreased in the presence of 0.9 g/l NH4Cl (e.g., 40% and 66%, respectively, in cells incubated for 95 h), while ammonium assimilation occurred via the glutamate dehydrogenase reaction. Furthermore, growth limitation occurred even in the presence of 0.5 g/l NH4Cl, when the DO concentration decreased from 135 to 30 microM. The activities of both CS and succinate dehydrogenase were dramatically decreased in cells grown at the lower DO concentration (e.g., 90% and 93% respectively, in a 95 h incubation), while ammonium assimilation was limited due to the low activities of both glutamate dehydrogenase and glutamate synthase. It is concluded that the threshold of ammonium concentration at which growth of A. lipoferum is limited, depends on the DO concentration in the medium.
Diazotrophic bacteria were isolated from the rhizosphere of field-grown Triticum aestivum, Hordeum vulgare, and Avena sativa grown in various regions of Greece. One isolate, with the highest nitrogen-fixation ability from each of the eleven rhizospheres, was selected for further characterisation. Diazotrophic strains were assessed for plant-growth-promoting traits such as indoleacetic acid production and phosphate solubilisation. The phylogenies of 16S rRNA gene of the selected isolates were compared with those based on dnaK and nifH genes. The constructed trees indicated that the isolates were members of the species Azospirillum brasilense, Azospirillum zeae, and Pseudomonas stutzeri. Furthermore, the ipdC gene was detected in all A. brasilence and one A. zeae isolates. The work presented here provides the first molecular genetic evidence for the presence of culturable nitrogen-fixing P. stutzeri and A. zeae associated with field-grown A. sativa and H. vulgare in Greece.
Olive fruits of the Greek variety “Amfissis” were stored under industrial conditions and the microorganisms of their microflora were isolated and identified. Selected molds of the microflora were screened for production of lipase and lipoxygenase, two enzymes that are known to contribute in the biogenesis of olive oil aroma compounds through the lipoxygenase pathway. Penicillium strain was identified as the most potent enzyme producer, the yields being 7300 U/L for lipase and 6.8 U/mg for lipoxygenase. Kinetic studies of enzyme production by Penicillium showed that lipase was mostly active at the beginning of growth, while lipoxygenase activity increased gradually and peaked at the end of growth. Next, the effect of the microflora on volatile biogenesis was evaluated by comparing the profiles of volatiles of virgin olive oil versus processed olive oil obtained by extracting olives that were processed by their natural microflora. The volatiles profile showed that the processed olive oil contained novel volatile compounds that could enhance its flavor, but also contained some off‐flavor compounds that could reduce its organoleptic characteristics and thus its quality. Our results show that olive microflora can contribute to the biogenesis of olive oil volatile compounds and thus some of its members could potentially be used to enhance the aroma of olive oil compounds.
In this study, Arthrospira platensis was grown in the presence of different glycerol concentrations (0.5–9 g/L) under three light intensities (5, 10 and 15 Klux) in semi-continuous mode and under non-axenic conditions. The aim of this study was to investigate the growth performance, the biomass biochemical composition and any interactions between A. platensis and bacteria that would potentially grow as well on glycerol. The results here show that glycerol did not have any positive effect on biomass production of A. platensis. In contrast, it was observed that by increasing glycerol concentration the growth performance of A. platensis was restricted, while a gradual increase of bacteria population was observed, which apparently outcompeted and repressed A. platensis growth. Chlorophyll fluorescence measurements (Quantum Yields) revealed that glycerol was not an inhibiting factor per se of photosynthesis. On the other hand, cyanobacterial biomass grown on glycerol displayed a higher content in proteins and lipids. Especially, protein productivity was enhanced around 15–35% with the addition of glycerol compared to the control. In distinction, carbohydrate and photosynthetic pigments (phycocyanin and chlorophyll-α) content decreased with the increase of glycerol concentration. The results here suggest that A. platensis did not utilize glycerol for biomass production but most probably as metabolic energy carrier towards synthesis of proteins and lipids, which are more energy consuming metabolites compared to carbohydrates. The study revealed that the addition of glycerol at amounts of 0.5–1.5 g/L could be a strategy to improve protein productivity by A. platensis.
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