Contamination of soils by heavy metals is one of the most serious environmental problems that increases the risk of toxic metal entry into the food chains. When heavy metals enter the soil, they are progressively converted to the insoluble form by reactions with soil components. A variety of mechanisms such as absorption, ion exchange, co-precipitation and complexation incorporates heavy metals into soil minerals or bounds them to various soil phases. Organic acids are natural compounds that are secreted from the root of the plant and can affect the solubility and uptake of heavy metals.
To evaluate the effects of plant growth promoting rhizobacteria (PGPR) on organic acids production and heavy metal uptake by different cabbage varieties, a factorial pot experiment with completely randomized design and three replications was performed under the greenhouse conditions. The factors included (a) rhizosphere soils of three varieties of cabbage [Brassica oleracea var. acephala L. (Ornamental cabbage), Brassica oleracea var. italica L. (Broccoli cabbage) and Brassica oleracea var. capitata L. (Cabbage)] and (b) five species of PGPR consisting of Pseudomonas putida PTCC 1694, Bacillus megaterium PTCC 1656, Proteus vulgaris PTCC 1079, Bacillus subtilis PTCC 1715 and Azotobacter chroococcum and control (without rhizobacteria) used to inoculate the rhizosphere soils. The experiment had 18 treatments and there were 54 experimental units and three seedlings of cabbage were planted in each pot. In all treatments inoculated with rhizobacterial species, 2 ml of a bacterial suspension with 107-108 (cfu ml-1) were used to inoculate the soil of root area. The data obtained in this study were statistically analyzed by SAS software (version 9.4) and the mean comparison was performed by Duncan’s multiple range test at 1 and 5 percent probability levels.
The analysis of variance (ANOVA) showed that the cabbage varieties, bacterial inoculation and their interactions had significant effects (p < 0.01) on organic acids concentration, fresh and dry biomass of plant, concentrations of Pb and Cd in root and shoot of cabbage plant. The results showed that inoculation of the rhizosphere soils with PGPR species increased organic acids concentration of rhizosphere. The highest concentration of malic and citric acids in rhizosphere soil (9.59 and 118.34 mg dl-1, respectively) was obtained when the rhizosphere soils of the broccoli were inoculated with Pseudomonas putida PTCC 1694 and the highest concentration of acetic acid in rhizosphere (233.88 mg dl-1) was determined when the rhizosphere of broccoli were inoculated with Bacillus megaterium PTCC 1656. Inoculation of the rhizosphere with PGPR species also increased the fresh and dry biomass of plant, and Pb and Cd concentrations in cabbage root and shoot. The highest fresh and dry biomass of cabbage (416.77 and 76.96 g in the plot, respectively) were obtained when the rhizosphere soils of cabbage were inoculated with Bacillus megaterium PTCC 1656, the highest concentration of Pb in the root and shoot and Cd in the root of cabbage (12.20, 90.77 and 9.01 mg kg-1, respectively) were obtained when the rhizosphere soils of the ornamental cabbage were inoculated with Pseudomonas putida PTCC 1694. Inoculation of the rhizosphere soils of the ornamental cabbage, broccoli and cabbage by B. megaterium PTCC1656 caused an increase in the DOC concentration by 137, 150 and 120%, respectively, compared to uninoculated rhizosphere soils. Bacterial inoculation also increased the concentrations of available phosphorus in the rhizosphere soils and the highest concentration of phosphorus was measured in the treatments inoculated by P. putida PTCC1694. Furthermore, the concentrations of available phosphorus in the rhizosphere soils of the ornamental cabbage, broccoli and cabbage increased by 79, 71 and 111%, respectively, relative to uninoculated rhizosphere soils.
It is concluded that inoculation of Pb and Cd contaminated soils by PGPR species, especially Bacillus megaterium PTCC 1656 and Pseudomonas putida PTCC 1694, enhances the tolerance of host plants, metal uptake performance and thus phytoremediation process by increasing the metal bioavailability and biomass production of the plant. As the distribution and accumulation of heavy metals in plant tissues are important factors for evaluation of plant role in phytoremediation of polluted soils, the PGPR inoculation of rhizosphere soils can be used as a biotechnological tool to enhance biomass production and plant uptake and thus the efficiency of phytoextraction.
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