Iron (Fe), an essential element for plant growth, is abundant in soil but with low bioavailability. Thus, plants developed specialized mechanisms to sequester the element. Beneficial microbes have recently become a favored method to promote plant growth through increased uptake of essential micronutrients, like Fe, yet little is known of their mechanisms of action. Functional mutants of the epiphytic bacterium Azospirillum brasilense, a prolific grass-root colonizer, were used to examine mechanisms for promoting iron uptake in Zea mays. Mutants included HM053, FP10, and ipdC, which have varying capacities for biological nitrogen fixation and production of the plant hormone auxin. Using radioactive iron-59 tracing and inductively coupled plasma mass spectrometry, we documented significant differences in host uptake of Fe2+/3+ correlating with mutant biological function. Radioactive carbon-11, administered to plants as 11CO2, provided insights into shifts in host usage of ‘new’ carbon resources in the presence of these beneficial microbes. Of the mutants examined, HM053 exhibited the greatest influence on host Fe uptake with increased plant allocation of 11C-resources to roots where they were transformed and exuded as 11C-acidic substrates to aid in Fe-chelation, and increased C-11 partitioning into citric acid, nicotianamine and histidine to aid in the in situ translocation of Fe once assimilated.

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Previous studies have shown that (i) Dal81p and Dal82p are required for allophanate-induced gene expression in Saccharomyces cerevisiae; (ii) the cis-acting element mediating the induced transcriptional response to allophanate is a dodecanucleotide, UIS ALL ; and (iii) Dal82p binds specifically to UIS ALL . Here we show that Dal82p is localized to the nucleus and parallels movement of the DNA through the cell cycle. Deletion analysis of DAL82 identified and localized three functional domains. Electrophoretic mobility shift assays identified a peptide (consisting of Dal82p amino acids 1-85) that is sufficient to bind a DNA fragment containing UIS ALL . LexA-tethering experiments demonstrated that Dal82p is capable of mediating transcriptional activation. The activation domain consists of two parts: (i) an absolutely required core region (amino acids 66 -99) and (ii) less well defined regions flanking residues 66 -99 that are required for full wild-type levels of activation. The Dal82p C terminus contains a predicted coiled-coil motif that down-regulates Dal82p-mediated transcriptional activation.

An increasing global population of over 4.5 billion people drives increasing demand for calories—30% of which are satisfied by grain crops, such as maize. High-density farming practices have been implemented but tend to deplete the soil of essential elements resulting in lower nutritional value, notably iron, of cultivated crops. Low iron content in staple crops can contribute over time to severe, even fatal, micronutrient deficiencies. Enhancing grain iron content using post-harvest biofortification strategies can be costly. However, field inoculation using biologics like Azospirillum brasilense (HM053) can be a cost-effective alternative to improving crop nutritional value. Using ion chromatography with chemiluminescence detection, we have shown that maize seeds harvested from outdoor pot-grown plants possessed a four-fold higher iron content as ferrous iron (Fe+2) compared to non-inoculated plants. Seeds from A. brasilense HM053-inoculated plants also contained approximately 13 nmol of ferritin per ground dried weight of kernel compared to 3 nmol from non-inoculated plants. In addition, A. brasilense HM053 inoculation increased crop yield 30–50% relative to non-inoculated plants.

Among the PGPB, the genus Azospirillum—with an emphasis on A. brasilense—is likely the most studied microorganism for mitigation of plant stress. Here, we report the investigation of functional mutants HM053, ipdC and FP10 of A. brasilense to understand how the biological functions of these microorganisms can affect host Zn uptake. HM053 is a Nif+ constitutively expressed strain that hyper-fixes N2 and produces high levels of the plant’s relevant hormone auxin. FP10 is a Nif- strain deficient in N2-fixation. ipdC is a strain that is deficient in auxin production but fixes N2. Zn uptake was measured in laboratory-based studies of 3-week-old plants using radioactive 65Zn2+ (t½ 244 days). Principal Component Analysis was applied to draw out correlations between microbial functions and host 65Zn2+ accumulation. Additionally, statistical correlations were made to our prior data on plant uptake of radioactive 59Fe3+ and 59Fe2+. These correlations showed that low microbial auxin-producing capacity resulted in the greatest accumulation of 65Zn. Just the opposite effect was noted for 59Fe where high microbial auxin-producing capacity resulted in the greatest accumulation of that tracer.