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Azospirillum brasilense is a plant growth promoting rhizobacterium (PGPR) that is being increasingly used in agriculture in a commercial scale. Recent research has elucidated key properties of A. brasilense that contribute to its ability to adapt to the rhizosphere habitat and to promote plant growth. They include synthesis of the auxin indole-3-acetic acid, nitric oxide, carotenoids, and a range of cell surface components as well as the ability to undergo phenotypic variation. Storage and utilization of polybetahydroxyalkanoate polymers are important for the shelf life of the bacteria in production of inoculants, products containing bacterial cells in a suitable carrier for agricultural use. Azospirillum brasilense is able to fix nitrogen, but despite some controversy, as judging from most systems evaluated so far, contribution of fixed nitrogen by this bacterium does not seem to play a major role in plant growth promotion. In this review, we focus on recent advances in the understanding of physiological properties of A. brasilense that are important for rhizosphere performance and successful interactions with plant roots., (© 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

Acidovorax citrulli is the causal agent of bacterial fruit blotch of cucurbits. We have shown previously that type IV pili (TFP) are required for wild-type levels of virulence of A. citrulli on melon and that this pathogen can colonize and move thorough the xylem vessels of host seedlings. Here, comparative studies between wild-type and TFP mutant strains using microfluidic flow chambers demonstrated that TFP play a critical role in both the surface attachment and the biofilm formation of A. citrulli under a medium flow. Additionally, TFP null mutants were unable to perform twitching movement against the direction of medium flow. Assays using a flagellin mutant showed that, in contrast to TFP, polar flagella do not contribute to the adhesion and biofilm formation of A. citrulli under tested conditions. Also, flagellum-mediated swimming motility of wild-type strains was not observed under medium flow. These results imply that TFP may play an important role in colonization and spread in the xylem vessels under sap flow conditions, while polar flagella could be more important for spread during periods of time when xylem flow is minimal., (© 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

Here we report the identification of a glycogen phosphorylase (glgP) gene in the plant growth-promoting rhizobacterium Azospirillum brasilense, Sp7, and the characterization of a glgP marker exchange mutant of this strain. The glgP mutant showed a twofold reduction of glycogen phosphorylase activity and an increased glycogen accumulation as compared with wild-type Sp7, indicating that the identified gene indeed encodes a protein with glycogen phosphorylase activity. Interestingly, the glgP mutant had higher survival rates than the wild type after exposure to starvation, desiccation and osmotic pressure. The mutant was shown to be compromised in its biofilm formation ability. Analysis of the exopolysaccharide sugar composition of the glgP mutant revealed a decrease in the amount of glucose, accompanied by increases in rhamnose, fucose and ribose, as compared with the Sp7 exopolysaccharide. To the best of our knowledge, this is the first study that demonstrates GlgP activity in A. brasilense, and shows that glycogen accumulation may play an important role in the stress endurance of this bacterium.

The response regulatory gene flcA controls the differentiation process of Azospirillum brasilense from vegetative state to cyst-like forms, both in culture and in association with plants. In contrast to the wild type parental strain Sp7, strain Sp72002, a Tn5 induced flcA(-) mutant, does not aggregate, does not differentiate from motile, vibroid cells into nonmotile, cyst-like forms and lacks some of the exopolysaccharide material on its cell surface. Here we aimed to identify differentially expressed genes whose expression could be modulated by flcA. Using cDNA-amplified fragment length polymorphism, we detected 81 transcript-derived fragments showing differential expression during exponential growth phase in an aggregation-inducing medium containing high C : N ratio. The fragments were sequenced and analyzed, and expression of ten genes encoding known proteins was compared between the two strains by reverse transcriptase polymerase chain reaction-(RT-PCR). RT-PCR analyses confirmed the differential patterns observed by cDNA-AFLP in nine of those genes. Among them are genes homologous to nodQ, involved in sulfation; narK, involved in nitrite/nitrate transport, and flp, involved in autoaggregation; as well as genes encoding a biopolymer transport protein, and the signal recognition particle. This work demonstrates the usefulness of the cDNA-AFLP approach to reveal genes that are differentially expressed during aggregation in Azospirillum brasilense and provides insights into the aggregation process of this bacterium.