Although salinity inhibits plant growth, application of appropriate rhizosphere bacteria can diminish this negative effect. We studied one possible mechanism that may underlie this beneficial response. Wheat plants were inoculated with Bacillus subtilis IB-22 and Pseudomonas mandelii IB-Ki14 and their consequences for growth, water relations, and concentrations of the hormone abscisic acid (ABA) were followed in the presence of soil salinity. Salinity alone increased ABA concentration in wheat leaves and roots and this was associated with decreased stomatal conductance, but also with chlorophyll loss. Bacterial treatment raised ABA concentrations in roots, suppressed accumulation of leaf ABA, decreased chlorophyll loss, and promoted leaf area and transpiration. However, water balance was maintained due to increased water uptake by inoculated plants, brought about in part by a larger root system. The effect may be the outcome of ABA action since the hormone is known to maintain root extension in stressed plants. Root ABA concentration was highest in salt-stressed plants inoculated with B. subtilis and this contributed to greater root hydraulic conductivity. We conclude that bacteria can raise salt resistance in wheat by increasing root ABA, resulting in larger root systems that can also possess enhanced hydraulic conductivity thereby supporting better-hydrated leaves.

The stomatal closure of salt-stressed plants reduces transpiration bringing about the maintenance of plant tissue hydration. The aim of this work was to test for any involvement of aquaporins (AQPs) in stomatal closure under salinity. The changes in the level of aquaporins in the cells were detected with the help of an immunohistochemical technique using antibodies against HvPIP2;2. In parallel, leaf sections were stained for abscisic acid (ABA). The effects of salinity were compared to those of exogenously applied ABA on leaf HvPIP2;2 levels and the stomatal and leaf hydraulic conductance of barley plants. Salinity reduced the abundance of HvPIP2;2 in the cells of the mestome sheath due to it being the more likely hydraulic barrier due to the deposition of lignin, accompanied by a decline in the hydraulic conductivity, transpiration, and ABA accumulation. The effects of exogenous ABA differed from those of salinity. This hormone decreased transpiration but increased the shoot hydraulic conductivity and PIP2;2 abundance. The difference in the action of the exogenous hormone and salinity may be related to the difference in the ABA distribution between leaf cells, with the hormone accumulating mainly in the mesophyll of salt-stressed plants and in the cells of the bundle sheaths of ABA-treated plants. The obtained results suggest the following succession of events: salinity decreases water flow into the shoots due to the decreased abundance of PIP2;2 and hydraulic conductance, while the decline in leaf hydration leads to the production of ABA in the leaves and stomatal closure.

Pseudomonas mandelii strain IB-Ki14 has recently been shown to strengthen the apoplastic barriers of salt-stressed plants, which prevents the entry of toxic sodium. It was of interest to find out whether the same effect manifests itself in the absence of salinity and how this affects the hydraulic conductivity of barley plants. Berberine staining confirmed that the bacterial treatment enhanced the deposition of lignin and suberin and formation of Casparian bands in the roots of barley plants. The calculation of hydraulic conductance by relating transpiration to leaf water potential showed that it did not decrease in bacteria-treated plants. We hypothesized that reduced apoplastic conductivity could be compensated by the higher conductivity of the water pathway across the membranes. This assumption was confirmed by the results of the immunolocalization of HvPIP2;5 aquaporins with specific antibodies, showing their increased abundance around the areas of the endodermis and exodermis of bacteria-treated plants. The immunolocalization with antibodies against auxins and abscisic acid revealed elevated levels of these hormones in the roots of plants treated with bacteria. This root accumulation of hormones is likely to be associated with the ability of Pseudomonas mandelii IB-Ki14 to synthesize these hormones. The involvement of abscisic acid in the control of aquaporin abundance and auxins—in the regulation of and formation of apoplast barriers—is discussed.

Increased auxin levels in root primordia are important in controlling root branching, while their interaction with abscisic acid (ABA) likely regulates lateral root development in water-deficient plants. The role of ABA accumulation in regulating root branching was investigated using immunolocalization to detect auxin (indoleacetic acid, IAA) and ABA (abscisic acid) in root primordia of the ABA-deficient barley mutant Az34 and its parental genotype (cv. Steptoe) barley plants. Osmotic stress strongly inhibited lateral root branching in Steptoe plants, but hardly affected Az34. Root primordial cells of Steptoe plants had increased immunostaining for ABA but diminished staining for IAA. ABA did not accumulate in root primordia of the Az34, and IAA levels and distribution were unaltered. Treating Az34 plants with exogenous ABA decreased root IAA concentration, while increasing root primordial ABA accumulation and decreasing root primordial IAA concentration. Although ABA treatment of Az34 plants increased the root primordial number, it decreased the number of visible emerged lateral roots. These effects were qualitatively similar to that of osmotic stress on the number of lateral root primordia and emerged lateral roots in Steptoe. Thus ABA accumulation (and its crosstalk with auxin) in root primordia seems important in regulating lateral root branching in response to water stress.

The role of reactive oxygen species (ROS) in ABA-induced increase in hydraulic conductivity was hypothesized to be dependent on an increase in aquaporin water channel (AQP) abundance. Single ABA application or its combination with ROS manipulators (ROS scavenger ascorbic acid and NADPH oxidase inhibitor diphenyleneiodonium chloride (DPI)) were studied on detached roots of barley plants. We measured the osmotically driven flow rate of xylem sap and calculated root hydraulic conductivity. In parallel, immunolocalization of ABA and HvPIP2;2 AQPs was performed with corresponding specific antibodies. ABA treatment increased the flow rate of xylem, root hydraulic conductivity and immunostaining for ABA and HvPIP2;2, while the addition of antioxidants prevented the effects of this hormone. The obtained results confirmed the involvement of ROS in ABA effect on hydraulic conductivity, in particular, the importance of H₂O₂ production by ABA-treated plants for the effect of this hormone on AQP abundance.

Little information is available on how rhizosphere bacteria affect abscisic acid (ABA) levels in plants and whether these bacterial effects are associated with improved plant water status. In this study, we tested the hypothesis that the stimulation of plant growth may be associated with the ability of ABA to increase the hydraulic conductivity of roots through the up-regulation of aquaporin. To do this, we studied the effect of bacteria capable of producing ABA on a barley mutant deficient in this hormone. Measurements of hydraulic conductivity of the ABA-deficient barley mutant Az34 showed that its tissues exhibited a reduced ability to conduct water, which correlated with lower ABA content in plants. The inoculation of Bacillus subtilis IB-22 stimulated the growth of both the mutant and its parent variety. Also, under the influence of bacteria, the ABA content in plants increased, and the increase was more significant in the mutant. This effect was accompanied by an increase in hydraulic conductivity in the roots of the ABA-deficient mutant, and immunolocalization using antibodies against PIP2;1 and PIP2;2 aquaporins revealed an increase in their abundance. Thus, the results obtained support the hypothesis about the importance of a sufficiently high ABA content in plants to maintain the abundance of aquaporins, hydraulic conductivity and the growth of barley plants.