Monitoring of Rhizosphere Bacterial Communities in Soil with Sewage Sludge Addition Using Two Molecular Fingerprinting Methods: Do These Methods Give Similar Results?

In this study, bacterial genetic diversity from the rhizosphere of barley and wheat were studied. The plants were sown in pots with aliquot amount of 15 t/ha concentration of soil additive derived from sewage sludge and agricultural by-products represented by wastes from grain mill industry and crushed corn cobs. The plants sown in pots without the addition of soil additive represented control samples. The rhizosphere samples were collected on two dates (plant flowering and maturity) and the composition of bacterial communities were detected using two molecular fingerprinting methods – automated ribosomal intergenic spacer analysis (ARISA) and terminal restriction fragment length polymorphism (T-RFLP). Microbial biomass expressed as the amount of metagenomics DNA was higher in soils with addition of soil additive, except during maturity stage in barley rhizosphere. Nevertheless, statistically significant differences between control and sludge samples were not detected in any case. Similarly, no changes were detected in the composition of bacterial community between control and sludge samples in barley and wheat rhizosphere by using cluster analysis. Only minor temporal changes in the composition of bacterial community between flowering and maturity periods were observed. These changes were related to the samples collected in the plant maturity stage. In this stage, plants were completely mature and their impact on the rhizosphere bacterial communities in the form of root exudates was limited. Statistically significant differences between ARISA and T-RFLP methods were detected in all measured values of diversity indices. Despite these differences, both methods gave results leading to similar conclusions.