Anaerobic Degradation of Naphthalene and Pyrene by Sulfate-Reducing Cultures Enriched from Former Manufactured Gas Plant Soil.

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) under completely anaerobic sulfate-reducing conditions is an energetically challenging process. To date, anaerobic degradations of only two-ringed naphthalene and three-ringed phenanthrene by sediment-free and enriched sulfate-reducing bacteria have been reported. In this study, sulfate-reducing enrichment cultures capable of degrading naphthalene and four-ringed PAH, pyrene, were enriched from a contaminated former gas plant site soil. Bacterial community composition analysis revealed that a naphthalene-degrading enrichment culture, MMNap, was dominated (84.90%) by a Gram-positive endospore-forming member of the genus Desulfotomaculum with minor contribution (8.60%) from a member of Clostridium. The pyrene-degrading enrichment, MMPyr, was dominated (97.40%) by a species of Desulfotomaculum. The sequences representing the Desulfotomaculum phylotypes shared 98.80% similarity to each other. After 150 days of incubation, MMNap degraded 195 µM naphthalene with simultaneous reduction of sulfate and accumulation of sulfide. Similarly, MMPyr degraded 114 µM pyrene during 180 days of incubation with nearly stochiometric sulfate consumption and sulfide accumulation. In both cases, the addition of sulfate reduction inhibitor, molybdate (20 mM), resulted in complete cessation of the substrate utilization and sulfate reduction that clearly indicated the major role of the sulfate-reducing Desulfotomaculum in biodegradation of the two PAHs. This study is the first report on anaerobic pyrene degradation by a matrix-free, strictly anaerobic, and sulfate-reducing enrichment culture.

Complete Genome Sequence of Rhodococcus sp. Strain 9, Isolated from Contaminated Australian Groundwater.

Here, we report the 7.7-Mbp genome sequence of Rhodococcus sp. strain 9, which was isolated from Australian groundwater contaminated with phenols and trichloroethylene. This strain has previously been shown to efficiently degrade p-nitrophenol and high-molecular-weight polycyclic aromatic hydrocarbons (PAHs).

Impact of Nitrate and Ammonium Concentrations on Co-Culturing of Tetradesmus obliquus IS2 with Variovorax paradoxus IS1 as Revealed by Phenotypic Responses.

Mutual interactions in co-cultures of microalgae and bacteria are well known for establishing consortia and nutrient uptake in aquatic habitats, but the phenotypic changes in terms of morphological, physiological, and biochemical attributes that drive these interactions have not been clearly understood. In this novel study, we demonstrated the phenotypic response in a co-culture involving a microalga, Tetradesmus obliquus IS2, and a bacterium, Variovorax paradoxus IS1, grown with varying concentrations of two inorganic nitrogen sources. Modified Bold's basal medium was supplemented with five ratios (%) of NO3-N:NH4-N (100:0, 75:25, 50:50, 25:75, and 0:100), and by maintaining N:P Redfield ratio of 16:1. The observed morphological changes in microalga included an increase in granularity and a broad range of cell sizes under the influence of increased ammonium levels. Co-culturing in presence of NO3-N alone or combination with NH4-N up to equimolar concentrations resulted in complete nitrogen uptake, increased growth in both the microbial strains, and enhanced accumulation of carbohydrates, proteins, and lipids. Total chlorophyll content in microalga was also significantly higher when it was grown as a co-culture with NO3-N and NH4-N up to a ratio of 50:50. Significant upregulation in the synthesis of amino acids and sugars and downregulation of organic acids were evident with higher ammonium uptake in the co-culture, indicating the regulation of carbon and nitrogen assimilation pathways and energy synthesis. Our data suggest that the co-culture of strains IS1 and IS2 could be exploited for effluent treatment by considering the concentrations of inorganic sources, particularly ammonium, in the wastewaters.

Extracellular Polymeric Substances Drive Symbiotic Interactions in Bacterial‒Microalgal Consortia.

The importance of several factors that drive the symbiotic interactions between bacteria and microalgae in consortia has been well realised. However, the implication of extracellular polymeric substances (EPS) released by the partners remains unclear. Therefore, the present study focused on the influence of EPS in developing consortia of a bacterium, Variovorax paradoxus IS1, with a microalga, Tetradesmus obliquus IS2 or Coelastrella sp. IS3, all isolated from poultry slaughterhouse wastewater. The bacterium increased the specific growth rates of microalgal species significantly in the consortia by enhancing the uptake of nitrate (88‒99%) and phosphate (92‒95%) besides accumulating higher amounts of carbohydrates and proteins. The EPS obtained from exudates, collected from the bacterial or microalgal cultures, contained numerous phytohormones, vitamins, polysaccharides and amino acids that are likely involved in interspecies interactions. The addition of EPS obtained from V. paradoxus IS1 to the culture medium doubled the growth of both the microalgal strains. The EPS collected from T. obliquus IS2 significantly increased the growth of V. paradoxus IS1, but there was no apparent change in bacterial growth when it was cultured in the presence of EPS from Coelastrella sp. IS3. These observations indicate that the interaction between V. paradoxus IS1 and T. obliquus IS2 was mutualism, while commensalism was the interaction between the bacterial strain and Coelastrella sp. IS3. Our present findings thus, for the first time, unveil the EPS-induced symbiotic interactions among the partners involved in bacterial‒microalgal consortia.

Exploring the Composition and Functions of Plastic Microbiome Using Whole-Genome Sequencing.

Besides the ecotoxicological consequences of microplastics and associated chemicals, the association of microbes on plastics has greater environmental implications as microplastics may select for unique microbiome participating in environmentally significant functions. Despite this, the functional potential of the microbiome associated with different types of plastics is understudied. Here, we investigate the interaction between plastic and marine biofilm-forming microorganisms through a whole-genome sequencing approach on four types of microplastics incubated in the marine environment. Taxonomic analysis suggested that the microplastic surfaces exhibit unique microbial profiles and niche partitioning among the substrates. In particular, the abundance of Vibrio alginolyticus and Vibrio campbellii suggested that microplastic pollution may pose a potential risk to the marine food chain and negatively impact aquaculture industries. Microbial genera involved in xenobiotic compound degradation, carbon cycling, and genes associated with the type IV secretion system, conjugal transfer protein TraG, plant-pathogen interaction, CusA/CzcA family heavy metal efflux transfer proteins, and TolC family proteins were significantly enriched on all the substrates, indicating the variety of processes operated by the plastic-microbiome. The present study gives a detailed characterization of the rapidly altering microbial composition and gene pools on plastics and adds new knowledge surrounding the environmental ramifications of marine plastic pollution.

Microalgal-bacterial consortia unveil distinct physiological changes to facilitate growth of microalgae.

Physiological changes that drive the microalgal-bacterial consortia are poorly understood so far. In the present novel study, we initially assessed five morphologically distinct microalgae for their ability in establishing consortia in Bold's basal medium with a bacterial strain, Variovorax paradoxus IS1, all isolated from wastewaters. Tetradesmus obliquus IS2 and Coelastrella sp. IS3 were further selected for gaining insights into physiological changes, including those of metabolomes in consortia involving V. paradoxus IS1. The distinct parameters investigated were pigments (chlorophyll a, b, and carotenoids), reactive oxygen species (ROS), lipids and metabolites that are implicated in major metabolic pathways. There was a significant increase (>1.2-fold) in pigments, viz., chlorophyll a, b and carotenoids, decrease in ROS and an enhanced lipid yield (>2-fold) in consortia than in individual cultures. In addition, the differential regulation of cellular metabolites such as sugars, amino acids, organic acids and phytohormones was distinct among the two microalgal-bacterial consortia. Our results thus indicate that the selected microalgal strains, T. obliquus IS2 and Coelastrella sp. IS3, developed efficient consortia with V. paradoxus IS1 by effecting the required physiological changes, including metabolomics. Such microalgal-bacterial consortia could largely be used in wastewater treatment and for production of value-added metabolites.

Metagenomics analysis identifies nitrogen metabolic pathway in bioremediation of diesel contaminated soil.

Nitrogen amendment is known to effectively enhance the bioremediation of hydrocarbon-contaminated soil, but the nitrogen metabolism in this process is not well understood. To unravel the nitrogen metabolic pathway(s) of diesel contaminated soil, six types of nitrogen sources were added to the diesel contaminated soil. Changes in microbial community and soil enzyme genes were investigated by metagenomics analysis and chemical analysis through a 30-day incubation study. The results showed that ammonium based nitrogen sources significantly accelerated the degradation of total petroleum hydrocarbon (TPH) (79-81%) compared to the control treatment (38%) and other non-ammonium based nitrogen amendments (43-57%). Different types of nitrogen sources could dramatically change the microbial community structure and soil enzyme gene abundance. Proteobacteria and Actinobacteria were identified as the two dominant phyla in the remediation of diesel contaminated soil. Metagenomics analysis revealed that the preferred metabolic pathway of nitrogen was from ammonium to glutamate via glutamine, and the enzymes governing this transformation were glutamine synthetase and glutamate synthetase; while in nitrate based amendment, the conversion from nitrite to ammonium was restrained by the low abundance of nitrite reductase enzyme and therefore retarded the TPH degradation rate. It is concluded that during the process of nitrogen enhanced bioremediation, the most efficient nitrogen cycling direction was from ammonium to glutamine, then to glutamate, and finally joined with carbon metabolism after transforming to 2-oxoglutarate.

Rhizodegradation of PAHs differentially altered by C3 and C4 plants.

Pyrosequencing of 16S ribosomal RNA (rRNA) was employed to characterize bacterial communities colonizing the rhizosphere of plants with C3 and C4 photosynthetic pathways grown in soil contaminated with polycyclic aromatic hydrocarbons (PAHs) after 60 and 120 days. The results of this study exhibited a clear difference in bacterial diversity between the rhizosphere and non-rhizosphere samples and between the rhizospheres of the C3 and C4 plants after 120 days. In both C3 and C4 rhizospheres, an incremental change in PAHs degrading bacterial genera was observed in the 120th day samples compared to the 60th day ones. Among the PAHs degrading bacterial genera, Pseudomonas showed good resistance to PAHs in the 120th day rhizosphere of both C3 and C4 plants. Conversely, the genus Sphingomonas showed sensitivity to PAHs in the 120th day rhizosphere soils of C3 plants only. Also, a significant increase in the PAHs degrading genera was observed at 120th day in the C4 rhizosphere in comparison to the C3 rhizosphere, which was reflected in a reduced PAHs concentration measured in the soil remediated with C4 plants rather than C3 plants. These results suggest that the rhizoremediation of PAHs was primarily governed by the plant photosystems, which led to differences in root secretions that caused the variation in bacterial diversity seen in the rhizospheres. This study is the first report to demonstrate the greater effectiveness of C4 plants in enhancing the PAHs degrading bacterial community than C3 plants.

Acid-adapted microalgae exhibit phenotypic changes for their survival in acid mine drainage samples.

Phenotypic plasticity or genetic adaptation in an organism provides phenotypic changes when exposed to the extreme environmental conditions. The resultant physiological and metabolic changes greatly enhance the organism's potential for its survival in such harsh environments. In the present novel approach, we tested the hypothesis whether acid-adapted microalgae, initially isolated from non-acidophilic environments, can survive and grow in acid-mine-drainage (AMD) samples. Two acid-adapted microalgal strains, Desmodesmus sp. MAS1 and Heterochlorella sp. MAS3, were tested individually or in combination (co-culture) for phenotypic changes during their growth in samples collected from AMD. The acid-adapted microalgae in AMD exhibited a two-fold increase in growth when compared with those grown at pH 3.5 in BBM up to 48 h and then declined. Furthermore, oxidative stress triggered several alterations such as increased cell size, granularity, and enhanced lipid accumulation in AMD-grown microalgae. Especially, the apparent limitation of phosphate in AMD inhibited the uptake of copper and iron in the cultures. Interestingly, growth of the acid-adapted microalgae in AMD downregulated amino acid metabolic pathways as a survival mechanism. This study demonstrates for the first time that acid-adapted microalgae can survive under extreme environmental conditions as exist in AMD by effecting significant phenotypic changes.

Sustainable Iron Recovery and Biodiesel Yield by Acid-Adapted Microalgae, Desmodesmus sp. MAS1 and Heterochlorella sp. MAS3, Grown in Synthetic Acid Mine Drainage.

Sustainable resource recovery is the key to manage the overburden of various waste entities of mining practices. The present study demonstrates for the first time a novel approach for iron recovery and biodiesel yield from two acid-adapted microalgae, Desmodesmus sp. MAS1 and Heterochlorella sp. MAS3, grown in synthetic acid mine drainage (SAMD). Virtually, there was no difference in the growth of the strain MAS3 both in Bold's basal medium (control) and SAMD. Using the IC50 level (200 mg L-1) and a lower concentration (50 mg L-1) of iron in SAMD, the cell granularity, exopolysaccharide (EPS) secretion, iron recovery, and biodiesel were assessed in both the strains. Both cell granularity and accumulation of EPS were significantly altered under metal stress in SAMD, resulting in an increase in total accumulation of iron. Growth of the microalgal strains in SAMD yielded 12-20% biodiesel, with no traces of heavy metals, from the biomass. The entire amount of iron, accumulated intracellularly, was recovered in the residual biomass. Our results on the ability of the acid-adapted microalgal strains in iron recovery and yield of biodiesel when grown in SAMD indicate that they could be the potential candidates for use in bioremediation of extreme habitats like AMD.

Endophyte-assisted phytoremediation: mechanisms and current application strategies for soil mixed pollutants.

Phytoremediation uses plants and associated microbes to remove pollutants from the environment and is considered a promising bioremediation method. Compared with well-described single contaminant treatments, the number of studies reporting phytoremediation of soil mixed pollutants has increased recently. Endophytes, including bacteria and fungi, exhibit beneficial traits for the promotion of plant growth, stress alleviation, and biodegradation. Moreover, endophytes either directly or indirectly assist host plants to survive high concentrations of organic and inorganic pollutants in the soil. Endophytic microorganisms can also regulate the plant metabolism in different ways, exhibiting a variety of physiological characteristics. This review summarizes the taxa and physiological properties of endophytic microorganisms that may participate in the detoxification of contaminant mixtures. Furthermore, potential biomolecules that may enhance endophyte mediated phytoremediation are discussed. The practical applications of pollutant-degrading endophytes and current strategies for applying this valuable bio-resource to soil phytoremediation are summarized.

Anaerobic Microbial Degradation of Polycyclic Aromatic Hydrocarbons: A Comprehensive Review.

Polycyclic aromatic hydrocarbons (PAHs) are a class of hazardous organic contaminants that are widely distributed in nature, and many of them are potentially toxic to humans and other living organisms. Biodegradation is the major route of detoxification and removal of PAHs from the environment. Aerobic biodegradation of PAHs has been the subject of extensive research; however, reports on anaerobic biodegradation of PAHs are so far limited. Microbial degradation of PAHs under anaerobic conditions is difficult because of the slow growth rate of anaerobes and low energy yield in the metabolic processes. Despite the limitations, some anaerobic bacteria degrade PAHs under nitrate-reducing, sulfate-reducing, iron-reducing, and methanogenic conditions. Anaerobic biodegradation, though relatively slow, is a significant process of natural attenuation of PAHs from the impacted anoxic environments such as sediments, subsurface soils, and aquifers. This review is intended to provide comprehensive details on microbial degradation of PAHs under various reducing conditions, to describe the degradation mechanisms, and to identify the areas that should receive due attention in further investigations.

Soil microalgae and cyanobacteria: the biotechnological potential in the maintenance of soil fertility and health.

The soil microbiota plays a major role in maintaining the nutrient balance, carbon sink, and soil health. Numerous studies reported on the function of microbiota such as plant growth-promoting bacteria and fungi in soil. Although microalgae and cyanobacteria are ubiquitous in soil, very less attention has been paid on the potential of these microorganisms. The indiscriminate use of various chemicals to enhance agricultural productivity led to serious consequences like structure instability, accumulation of toxic contaminants, etc., leading to an ecological imbalance between soil, plant, and microbiota. However, the significant role of microalgae and cyanobacteria in crop productivity and other potential options has been so far undermined. The intent of the present critical review is to highlight the significance of this unique group of microorganisms in terms of maintaining soil fertility and soil health. Beneficial soil ecological applications of these two groups in enhancing plant growth, establishing interrelationships among other microbes, and detoxifying chemical agents such as insecticides, herbicides, etc. through mutualistic cooperation by synthesizing enzymes and phytohormones are presented. Since recombinant technology involving genomic integration favors the development of useful traits in microalgae and cyanobacteria for their potential application in improvement of soil fertility and health, the merits and demerits of various such advanced methodologies associated in harnessing the biotechnological potential of these photosynthetic microorganisms for sustainable agriculture were also discussed.

Ecological role of Acinetobacter calcoaceticus GSN3 in natural biofilm formation and its advantages in bioremediation.

This study assessed the role of a new Acinetobacter calcoaceticus strain, GSN3, with biofilm-forming and phenol-degrading abilities. Three biofilm reactors were spiked with activated sludge (R1), green fluorescent plasmid (GFP) tagged GSN3 (R2), and their combination (R3). More than 99% phenol removal was achieved during four weeks in R3 while this efficiency was reached after two and four further operational weeks in R2 and R1, respectively. Confocal scanning electron microscopy revealed that GSN3-gfp strains appeared mostly in the deeper layers of the biofilm in R3. After four weeks, almost 7.07 × 107 more attached sludge cells were counted per carrier in R3 in comparison to R1. Additionally, the higher numbers of GSN3-gfp in R2 were unable to increase the efficiency as much as measured in R3. The presence of GSN3-gfp in R3 conveyed advantages, including enhancement of cell immobilization, population diversity, metabolic cooperation and ultimately treatment efficiency.

Bioremediation of soil long-term contaminated with PAHs by algal-bacterial synergy of Chlorella sp. MM3 and Rhodococcus wratislaviensis strain 9 in slurry phase.

Remediation of soil contaminated with pollutants using biological agents is more a sustainable and greener approach as compared to physico-chemical technologies. We recently confirmed that a microalga, Chlorella sp. MM3, and a bacterium, Rhodococcus wratislaviensis strain 9, can degrade high-molecular weight PAHs. In this study, an algal-bacterial system of these two strains was developed by long-term growth on a mixture of phenanthrene, pyrene, and benzo[a]pyrene (BaP). In a soil spiked with 50 mg L-1 phenanthrene, 10 mg L-1 of pyrene and 10 mg L-1 of BaP, the algal-bacterial system degraded these PAHs almost completely in slurry phase within 30 days. Also, the algal-bacterial system was able to successfully remediate these three PAHs in a soil long-term contaminated with 245.1 mg kg-1 of 16 PAHs and several heavy metals under slurry phase in 21 days. Use of such appropriate assays as chlorophyll estimation for the microalga and semi-quantitative PCR for the bacterium confirmed survival of both the strains during soil bioremediation. Moreover, the residual toxicity test involving Escherichia coli DH5α that expresses green fluorescent protein indicated the successful bioremediation of PAHs-contaminated soil in slurry phase. For the first time, here we demonstrate the great potential of an algal-bacterial synergy in bioremediation of soil long-term contaminated with PAHs even in the presence of toxic heavy metals.

Advances in the technologies for studying consortia of bacteria and cyanobacteria/microalgae in wastewaters.

The excessive generation and discharge of wastewaters have been serious concerns worldwide in the recent past. From an environmental friendly perspective, bacteria, cyanobacteria and microalgae, and the consortia have been largely considered for biological treatment of wastewaters. For efficient use of bacteria‒cyanobacteria/microalgae consortia in wastewater treatment, detailed knowledge on their structure, behavior and interaction is essential. In this direction, specific analytical tools and techniques play a significant role in studying these consortia. This review presents a critical perspective on physical, biochemical and molecular techniques such as microscopy, flow cytometry with cell sorting, nanoSIMS and omics approaches used for systematic investigations of the structure and function, particularly nutrient removal potential of bacteria‒cyanobacteria/microalgae consortia. In particular, the use of specific molecular techniques of genomics, transcriptomics, proteomics metabolomics and genetic engineering to develop more stable consortia of bacteria and cyanobacteria/microalgae with their improved biotechnological capabilities in wastewater treatment has been highlighted.

Acid-tolerant microalgae can withstand higher concentrations of invasive cadmium and produce sustainable biomass and biodiesel at pH 3.5.

Two acid-tolerant microalgae, Desmodesmus sp. MAS1 and Heterochlorella sp. MAS3, originally isolated from non-acidophilic environment, were tested for their ability to withstand higher concentrations of an invasive heavy metal, cadmium (Cd), at an acidic pH of 3.5 and produce biomass rich in biodiesel. The growth analysis, in terms of chlorophyll, revealed that strain MAS1 was tolerant even to 20 mg L-1 of Cd while strain MAS3 could withstand only up to 5 mg L-1. When grown in the presence of 2 mg L-1, a concentration which is 400-fold higher than that usually occurs in the environment, the microalgal strains accumulated >58% of Cd from culture medium at pH 3.5. FTIR analysis of Cd-laden biomass indicated production of significant amounts of biodiesel rich in fatty acid esters. This is the first study that demonstrates the capability of acid-tolerant microalgae to grow well and remove Cd at acidic pH.

Role of 2-mercaptopyridine on control of microbial influenced corrosion of copper CW024A metal in cooling water system.

In this present study, the biocorrosion behaviour of Bacillus thuringiensis EN2 and B. oleronius EN9 on copper metal CW024A (Cu) in cooling water system (1% chloride) were evaluated using weight loss, electrochemical impedance spectroscopy (EIS) and surface analysis. In presence of EN2 and EN9, the corrosion rates (CR) were higher, about 0.021 mm/y and 0.032 mm/y than control system (0.004 mm/y). On the other hand, the presence of corrosion inhibitor 2-mercaptopyridine (2-MCP) with bacteria (EN2 and EN9), the biofilm on metal surface was highly inhibited and thus reduces the corrosion rate (CR: 0.004 mm/y). The electrochemical behaviour of CW024A metal was correlated with the adsorbed corrosion inhibitor film and biofilm. Atomic force microscopy (AFM) analysis revealed that the presence of EN2 and EN9 more pits was observed on the metal surface rather than 2-MCP system. EIS confirms the inhibitor act as cathodic type of inhibitor and thus leads to the inhibition of CR. Overall, this work concluded that corrosion inhibitor (2-MCP) inhibits, the bacterial biofilm formation on the metal surface due to the formation of productive layer on metal surface as coordination of NH bond. Which leads to the reduction of bacterial attachment and thus higher corrosion inhibition efficiency (75%) obtained. This is the first work disclosing the role of 2-MCP formulations as potent anti-bacterial and corrosion inhibition efficiency on copper metal in cooling water tower environment.

Potential of acid-tolerant microalgae, Desmodesmus sp. MAS1 and Heterochlorella sp. MAS3, in heavy metal removal and biodiesel production at acidic pH.

Metals in traces are vital for microalgae but their occurrence at high concentrations in habitats is a serious ecological concern. We investigated the potential of two acid-tolerant microalgae, Desmodesmus sp. MAS1 and Heterochlorella sp. MAS3, isolated from neutral environments, for simultaneous removal of heavy metals such as copper (Cu), iron (Fe), manganese (Mn) and zinc (Zn), and production of biodiesel when grown at pH 3.5. Excepting Cu, the selected metals at concentrations of 10-20 mg L-1 supported good growth of both the strains. Cellular analysis for metal removal revealed the predominance of intracellular mechanism in both the strains resulting in 40-80 and 40-60% removal of Fe and Mn, respectively. In-situ transesterification of biomass indicated enhanced biodiesel yield with increasing concentrations of metals suggesting that both these acid-tolerant microalgae may be the suitable candidates for simultaneous remediation, and sustainable biomass and biodiesel production in environments like metal-rich acid mine drainages.

Metabolomics reveals defensive mechanisms adapted by maize on exposure to high molecular weight polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons are an important group of persistent organic pollutants. Using plants to remediate PAHs has been recognized as a cost-effective and environmentally friendly technique. However, the overall impact of PAHs on the regulation of plant metabolism has not yet been explored. In this study, we analyzed the alteration in the maize (Zea mays L.) metabolome on exposure to high molecular weight PAHs such as benzo[a]pyrene (BaP) and pyrene (PYR) in a hydroponic medium, individually and as a mixture (BaP + PYR) using GC-MS. The differences in the metabolites were analyzed using XCMS (an acronym for various forms (X) of chromatography-mass spectrometry), an online-based data analysis tool. A significant variation in metabolites was observed between treatment groups and the unspiked control group. The univariate, multivariate and pathway impact analysis showed there were more significant alterations in metabolic profiles between individual PAHs and the mixture of BaP and PYR. The marked changes in the metabolites of galactose metabolism and aminoacyl tRNA biosynthesis in PAHs treated maize leaves exhibit the adaptive defensive mechanisms for individual and PAHs mixture. Therefore, the metabolomics approach is essential for an understanding of the complex biochemical responses of plants to PAHs contaminants. This knowledge will shed new light in the field of phytoremediation, bio-monitoring, and environmental risk assessment.