Abstract. Pilot experiments were conducted to analyse the effect of different environmental factors on the rhizoremediation of petroleum-contaminated soil. Different plant species (cotton, ryegrass, tall fescue and alfalfa), the addition of fertilizer, different concentrations of total petroleum hydrocarbons (TPH) in the soil, bioaugmentation with effective microbial agents (EMA) and plant growth-promoting rhizobacteria (PGPR) and remediation time were tested as influencing factors during the bioremediation process of TPH. The results show that the remediation process can be enhanced by different plant species. The order of effectiveness of the plants was the following: tall fescue > ryegrass > alfalfa > cotton. The degradation rate of TPH increased with increased fertilizer addition, and a moderate urea level of 20 g N (Nitrogen)/m 2 was best for both plant growth and TPH remediation. A high TPH content is toxic to plant growth and inhibits the degradation of petroleum hydrocarbons. The results showed that a 5% TPH content gave the best degradation in soil planted with ryegrass. Bioaugmentation with different bacteria and PGPR yielded the following results for TPH degradation: cotton+EMA+PGPR > cotton+EMA > cotton+PGPR > cotton > control. Rapid degradation of TPH was found at the initial period of remediation caused by the activity of microorganisms. A continuous increase of degradation rate was found during the 30-90 days period followed by a slow increase during the 90-150 days period. These results suggest that rhizoremediation can be enhanced with the proper control of different influencing factors that affect both plant growth and microbial activity in the rhizosphere environment.

The soil microbiome consists of a vast variety of microorganisms which contribute to essential ecosystem services including nutrient recycling, protecting soil structure, and pathogen suppression. Recalcitrant organic compounds present in soils contaminated with fuel oil can lead to a decrease in functional redundancy within soil microbiomes. Ecopiling is a passive bioremediation technique involving biostimulation of indigenous hydrocarbon degraders, bioaugmentation through inoculation with known petroleum-degrading consortia, and phytoremediation. The current study investigates the assemblage of soil microbial communities and pollutant-degrading potential in soil undergoing the Ecopiling process, through the amplicon marker gene and metagenomics analysis of the contaminated soil. The analysis of key community members including bacteria, fungi, and nematodes revealed a surprisingly diverse microbial community composition within the contaminated soil. The soil bacterial community was found to be dominated by Alphaproteobacteria (60–70%) with the most abundant genera such as Lysobacter, Dietzia, Pseudomonas, and Extensimonas. The fungal community consisted mainly of Ascomycota (50–70% relative abundance). Soil sequencing data allowed the identification of key enzymes involved in the biodegradation of hydrocarbons, providing a novel window into the function of individual bacterial groups in the Ecopile. Although the genus Lysobacter was identified as the most abundant bacterial genus (11–46%) in all of the contaminated soil samples, the metagenomic data were unable to confirm a role for this group in petrochemical degradation. Conversely, genera with relatively low abundance such as Dietzia (0.4–9.0%), Pusillimonas (0.7–2.3%), and Bradyrhizobium (0.8–1.8%) did possess genes involved in aliphatic or aromatic compound degradation.