The Effects of Plant Growth Promoting Rhizobacteria on the Activity of some Enzymes in Rizosphere Soils under Lead and Cadmium Stress

Contamination of soils with heavy metals is one of the most serious environmental problems increasing the risk of the entry of heavy metals into food chains. Rhizosphere soil is distinct from the bulk soil and is defined as the volume of soil around living roots which is influenced by root activities. Enzymes are produced by both roots and soil microorganisms to alter nutrient availability in rhizosphere soil. Soil enzymes promote the transformation of matter and energy in the soil, and their activity has a close relationship with soil nutrient availability. Detection of microbial enzymes in a natural environment is important to understand biochemical activities and to verify the biotechnological potential of microorganisms. However, there are few reports to indicate the biotechnological potential of plant growth promoting rhizobacteria (PGPR) and their effects on the activity of bacterial enzymes in rhizosphere soils under the stress of heavy metals. Thus, in the present study lead and cadmium contaminated rhizosphere soils were inoculated with PGPR species to investigate the influence of these bacteria on the activity of some enzymes.  A factorial pot experiment with completely randomized design base and three replications was performed in the greenhouse conditions. The factors examined were (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 Pseudomonas putida PTCC 1694, Bacillus megaterium PTCC 1656, Proteus vulgaris PTCC 1079, Bacillus subtilis PTCC 1715 and Azotobacter chroococcum, used to inoculate the rhizosphere soils. There was also a control treatment (without rhizobacteria). The experiment had 18 treatments and there were 54 experimental units. To study rhizosphere soils, several rhizoboxes were used and three seedlings of cabbage were planted in the central part of each rhizobox (rhizosphere area). In treatments inoculated with rhizobacterial species, 2 ml of a bacterial suspension with 107-108 (cfu ml-1) was used to inoculate the soil of rootzone. After three months, cabbage varieties were harvested and the activity of alkaline phosphatase, acid phosphatase, urease, and dihydrogenase were measured in rhizosphere soils. The data obtained from this study were statistically analyzed by SPSS statistical software package (Version 9.4) and the variance of the data was analyzed by one-way ANOVAs (Duncan’s test) range test at 1 and 5 percent probability levels. The analysis of variance of the data (ANOVA) showed that the cabbage varieties, inoculation with PGPR species and their interactions had significant effects (p < 0.01) on the activity of alkaline phosphatase, acid phosphatase, urease, and dihydrogenase in rhizosphere soils. The results showed that inoculation of the rhizosphere soils with PGPR species increased the activity of soil enzymes. The highest activity of alkaline phosphatase (1529.28 µg pNP.g-1 dm.h-1) was measured in rhizosphere soils of the broccoli inoculated with Pseudomonas putida PTCC 1694. But, the highest activity of acid phosphatase (497.92 µg pNP.g-1dm.h-1) was obtained in rhizosphere soils of cabbage inoculated with Pseudomonas putida PTCC 1694. Also, the highest activity of urease (208.36 µg N-NH4+.g-1dm.2h-1) was observed in rhizosphere soils of the cabbage inoculated with Azotobacter chroococcum and the highest activity of dihydrogenase (8.71 µg TPF.g-1dm.16h-1) was observed when rhizosphere soils of the cabbage were inoculated with Bacillus subtilis PTCC1715. From the results of this study, it may be concluded that inoculation of Pb and Cd contaminated soils with PGPR species could modulate the toxic effects of heavy metals on plant and increase the activity of some key enzymes for plant growth in rhizosphere soils.