Phyllospheric bacteria were isolated from microsites around essential-oil-containing glands of two oregano (Origanum vulgare subsp. hirtum) lines. These bacteria, 20 isolates in total, were subjected to bioassays to examine their growth potential in the presence of essential oils at different concentrations. Although there were qualitative and quantitative differences in the essential oil composition between the two oregano lines, no differences were recorded in their antibacterial activity. In disk diffusion bioassays, four of the isolated strains could grow almost unrestrained in the presence of oregano oil, another five proved very sensitive, and the remaining 11 showed intermediate sensitivity. The strain least inhibited by oregano essential oil was further identified by complete16s rRNA gene sequencing as Pseudomonas putida. It was capable of forming biofilms even in the presence of oregano oil at high concentrations. Resistance of P. putida to oregano oil was further elaborated by microwell dilution bioassays, and its topology on oregano leaves was studied by electron microscopy. When inoculated on intact oregano plants, P. putida was able not only to colonize sites adjacent to essential oil-containing glands, but even to grow intracellularly. This is the first time that such prolific bacterial growth inside the glands has been visually observed. Results of this study further revealed that several bacteria can be established on oregano leaves, suggesting that these bacteria have attributes that allow them to tolerate or benefit from oregano secondary metabolites.

The reduction potential of the highly toxic Cr(VI) to the inert Cr(III) in an alkaline soil was studied during a 50-day experiment with Portulacaoleracea grown in pots. We aimed at assessing whether our test species can be a phytoremediation candidate for Cr(VI)-contaminated soils. We measured the Cr(VI) reduction rate in soil, determined the Cr(VI) and Cr(III) concentrations in aerial and root P. oleracea tissues, calculated the Transfer coe cient (TC=metal in plant over metal in soil) and the Translocation factor (TF=metal in aerial biomass over metal in roots) in order to assess Cr(VI) uptake and distribution in plant tissues, while we also studied the effect of added nitrogen in the studied parameters. We added ve different Cr(VI) levels,reaching 148.6 mg Cr(VI) kg -1 soil and also had two N levels (equivalent to 0 and 200kg ha -1 ). The results indicated that Cr in plant tissues was mainly found in its reduced form (Cr(III)) and only a minor fraction of Cr was detected in its oxidized form (Cr(VI)). The main remediation mechanism was found to be that of the naturally occurring Cr(VI) reduction that effectively produced Cr(III), followed by the uptake of Cr(VI) from our test plants. We also found that Cr(VI) in P. oleracea tissues was mainly found in roots and relatively low Cr(VI) concentrations were found in the above ground tissues. We concluded that P. oleraceais a tolerant plant species, especially if assisted with a su cient level of N fertilization, although it failed to approach the threshold of being categorized as an accumulator species.However, before reaching more conclusive suggestionsabout P. oleracea as a potential phytoremediation species, further investigation is necessary.

In a pot experiment, we assessed the potential of purslane (Portulaca oleracea) as a phytoremediation species in Cr(VI)-contaminated soils. We focused on the evaluation of phytotoxic Cr(VI) effects at concentrations reaching 150 mg Cr(VI) kg−1 and the possible stress amelioration effect of nitrogen on Cr(VI)-stressed plants. Treatments were T-0 (control), T-1 (25 mg Cr(VI) kg−1), T-2 = 50 mg kg−1, T-3 = 100 mg kg−1, and T-4 = 150 mg kg−1. We measured Cr(VI) concentration in aerial and root tissues, a series of parameters related to photosynthesis and plant growth, phosphorus aerial plant tissue content, and we also calculated indices (ratios) related to leaf growth and above ground tissue water content. Cr(VI) almost exclusively was found in root tissues; all physiological and growth parameters studied were severely affected and plants selectively accumulated phosphorus in aerial plant tissues with increasing Cr(VI) soil concentrations. On the other hand, N amendment resulted in improved plant features in some of the measured parameters: chlorophyll index was improved with added N at T-2, plant height was significantly higher at T-0, T-1, and T-2, and aerial dry weight and leaf area was higher at T-0; these effects indicate that added N did increase P. oleracea potential to ameliorate Cr(VI) toxic effects. We conclude that purslane showed a potential as a possible species to be successfully introduced to Cr(VI)-laden soils, but more research is certainly necessary.

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Agricultural lands, especially those where wheat is cultivated, in the vicinity of intense anthropogenic activities may be laden with potentially toxic elements (PTEs), resulting in increased risk for human health. In this study we monitored three regions located in central Greece, currently cultivated with wheat: Domokos and Eretria, two areas with abandoned chromium mines, but never studied before, and the industrial area of Volos, near a major steel factory. All soils were alkaline with medium CaCO3 content. As expected, Cr was extremely high in the first two areas (705.2 in Eretria and 777.5 mg kg−1 in Domokos); Ni was also found elevated (1227 in Eretria, 1315 in Domokos and 257.6 mg kg−1 in the steel factory), while other harmful metals (Cd, Cu, Pb and Zn) were rather low. As a result, pollution load index, a cumulative index showing the contamination level of an area, was higher than 1.0 in all three areas (Eretria = 2.20, Domokos = 2.28, and steel factory = 1.61), indicating high contamination and anthropogenic inputs. As for the wheat parts (shoots and grains), they were found to have no elevated concentrations of any of the measured metals in all three study areas, probably due to the alkaline soil pH that decelerates metal mobility. This was also confirmed by the very low soil-to-plant transfer coefficient values for all metals. In assessing the possible risk concerning human health, we found that the soil-to-human pathway would induce no significant risk (exhibited by hazard index of less than 1.0), while the risk from grain-to-human resulted in considerable risk for human health in the steel factory of Volos (where HI > 1.0). Our findings suggest that rural areas never studied before with a history in some offensive anthropogenic activity can prove to be a contamination hotspot; we regard this study as a pivotal for similarly never-visited-before areas casually cultivated with wheat (or other important crops for human nutrition). We further recognize the need for a more in-depth study that would acknowledge the geochemical speciation of the studied metals and also monitor other important crops and their possible uptake of PTEs.