Plant roots support the growth and activities of a wide variety of microorganisms that may have a profound effect on the growth and/or health of plants. Among these microorganisms, a high diversity of bacteria have been identified and categorized as deleterious, beneficial, or neutral with respect to the plant. The beneficial bacteria, termed plant growth-promoting rhizobacteria (PGPR), are widely studied by microbiologists and agronomists because of their potential in plant production. Azospirillum, a genus of versatile PGPR, is able to enhance the plant growth and yield of a wide range of economically important crops in different soils and climatic regions. Plant beneficial effects of Azospirillum have mainly been attributed to the production of phytohormones, nitrate reduction, and nitrogen fixation, which have been subject of extensive research throughout the years. These elaborate studies made Azospirillum one of the best-characterized genera of PGPR. However, the genetic and molecular determinants involved in the initial interaction between Azospirillum and plant roots are not yet fully understood. This review will mainly highlight the current knowledge on Azospirillum plant root interactions, in the context of preceding and ongoing research on the association between plants and plant growth-promoting rhizobacteria.

An antimicrobial compound was isolated from Azospirillum brasilense culture extracts by high-performance liquid chromatography and further identified by gas chromatography-mass spectrometry as the auxin-like molecule, phenylacetic acid (PAA). PAA synthesis was found to be mediated by the indole-3-pyruvate decarboxylase, previously identified as a key enzyme in indole-3-acetic acid (IAA) production in A. brasilense. In minimal growth medium, PAA biosynthesis by A. brasilense was only observed in the presence of phenylalanine (or precursors thereof). This observation suggests deamination of phenylalanine, decarboxylation of phenylpyruvate, and subsequent oxidation of phenylacetaldehyde as the most likely pathway for PAA synthesis. Expression analysis revealed that transcription of the ipdC gene is upregulated by PAA, as was previously described for IAA and synthetic auxins, indicating a positive feedback regulation. The synthesis of PAA by A. brasilense is discussed in relation to previously reported biocontrol properties of A. brasilense.Azospirillum is a well-studied genus of plant growth-promoting bacteria (PGPB), which colonizes the rhizosphere of numerous crop plants in tropical and subtropical regions (5, 6). Different mechanisms, such as phytohormone production, nitrate reduction, and nitrogen fixation, have been proposed to explain improved plant growth following inoculation with Azospirillum (5,7,9,22,37,38,53). The production of phytohormones, and more specifically the auxin indole-3-acetic acid (IAA), has been recognized as an important factor in direct plant-growth-promoting abilities of A. brasilense (5,18,19,37).Azospirillum sp. are not typical biocontrol agents of soilborne plant pathogens (5). Apart from some reports on bacteriocins and siderophores, no other antibacterial substances in Azospirillum sp. have been identified so far (39,40,50,55,60). However, there have been reports of moderate biocontrol capabilities of Azospirillum brasilense against crown gall disease, bacterial leaf blight of mulberry, and bacterial leaf and/or vascular diseases of tomato (1,3,4,46,54). In addition, A. brasilense can restrict the proliferation of other nonpathogenic rhizosphere bacteria (21). Nevertheless, the exact mechanisms involved in Azospirillum acting as a putative biocontrol agent are not yet known. Some reports therefore indicate that the protective mechanism may be indirectly explained by the plant growth promotion effect or by outcompeting other bacteria hosted by the same plant (3,46).In the present study, we attempted to further screen A. brasilense supernatant (extracts) for the presence of metabolites (besides IAA), which may be involved in the persistence of Azospirillum in the rhizosphere. This screening led to the identification of phenylacetic acid (PAA), an auxin-like molecule with antimicrobial activity. MATERIALS AND METHODSStrains, plasmids, media, and culture conditions. Strains and plasmids used in this study are listed in Table 1. A. brasilense was grown at 30°C in Luria-Bertani (LB) med...

Nitrogen regulation in bacteria involves the capacity to sense the availability of fixed nitrogen and to translate a signal indicating nitrogen deficiency or nitrogen excess into a cellular response. One of the key enzymes in this complex regulation process, the uridylyltransferase/uridylyl-removing (UTase) enzyme, encoded by the glnD gene, was characterised in the diazotroph Azospirillum brasilense, which promotes plant growth. The glnD gene product is responsible for the uridylylation of both P(II)-like nitrogen regulatory proteins, P(II) and P(Z), depending on the nitrogen status of the cell. The nitrogen-regulated activity of the main ammonium-assimilating enzyme, glutamine synthetase, is not altered in a glnD-Tn 5-B30 insertion mutant. UTase influences processes that are regulated by the NtrB-NtrC two-component histidine protein kinase system, such as ammonium uptake and nitrate assimilation. Moreover, the glnD gene product is indispensable for the activation of nitrogen fixation. Transcription of glnD is up-regulated under nitrogen-fixing conditions. This regulation is only partially dependent on the global nitrogen regulation (Ntr) system.

The salAB genes of Azospirillum irakense KBC1, which encode two aryl-beta-glucosidases, are required for growth on salicin. In the 4-kb region upstream of the salAB genes, two additional genes, salC and salR, were identified. SalC shows characteristics of the outer membrane receptors in the FepA/FhuA family. The salC AB genes are transcribed as a polycistronic mRNA. The salR gene encodes a protein homologous to the LacI/GalR family of transcriptional repressors. Expression of the sal operon, measured by means of a salC-gusA translational fusion in A. irkense KBC1, requires the presence of aryl-beta-glucosides such as arbutin and salicin. Expression is markedly enhanced when a simple carbon source, like glucose, cellobiose or malate, is added to the medium. In a salR mutant, expression of the salC-gusA fusion does not require an aryl-beta-glucoside inducer. Expression of a salR-gusA fusion is constitutive. The product of arbutin hydrolysis (hydroquinone) partly inhibits the expression of a salC-gusA fusion in arbutin- or salicin-containing minimal medium. This effect is independent of SalR. Salicylalcohol, the hydrolysis product of salicin, also partly inhibits salC expression in a SalR-independent fashion, but only in salicin-containing minimal medium.

No abstract