Arbuscular mycorrhizal fungus inocula from coastal sand dunes arrest olive cutting growth under salinity stress.

Cultivation of olive trees covers large coastal areas of land in Mediterranean regions, many of them characterized by low soil fertility and exposed to salinity and seasonal drought. In this frame, we developed mixed community inocula of arbuscular mycorrhizal fungi (AMF) derived from the extreme, seasonally arid environments of six Mediterranean sand dunes and evaluated their effects, in the form of community inocula, on rooted semi-woody olive tree cuttings (Olea europaea cv. Koroneiki). The plantlets were grown in the greenhouse for 10 months under 50 mM and 100 mM concentrations of NaCl, successively applied to induce osmotic stress. Inoculation had a positive effect on plant growth and nutrient uptake. However, the three best-performing inocula in early colonization and in plant growth enhancement also resulted in high plant sensitivity to high salinity, which was not observed for the other three inocula. This was expressed by decreased nutrient uptake and drastically lower plant growth, plant photosynthesis, and stomatal conductance (generally an over 50% reduction compared to no salinity application). Amplicon sequencing analysis of the olive plants under salinity stress showed that the AMF communities in the roots were clearly differentiated by inoculation treatment. We could not, however, consistently associate the plant responses observed under high salinity with specific shared AMF community membership or assembly attributes. The observed physiological overreaction to osmotic stress may be an adaptation trait, potentially brought about by host selection coupled to abiotic environmental filtering, in the harsh conditions from which the AMF inocula were derived. The overreaction may, however, be undesirable if conveyed to allochthonous plants at an agronomic level.

Strong host-specific selection and over-dominance characterize arbuscular mycorrhizal fungal root colonizers of coastal sand dune plants of the Mediterranean region.

Sand dunes of the Mediterranean region constitute drought-stressed, low-fertility ecosystems. Arbuscular mycorrhizal fungi (AMF) are regarded as key components of their biota, that contribute to plant host adaptation and fitness. However, AMF community assembly rules in the roots of the psammophilous plants of coastal sand dunes have not been investigated. We studied the root colonizing AMF communities of four characteristic native plants of eastern Mediterranean coastal foredunes, in nine locations in Greece. Host-specificity (plant identity) was the major driver of AMF community assembly in the plant roots, while geographical distance between locations was not related to differences in the AMF communities. Additionally, colonizer AMF communities were characterized by over-dominance of a single operational taxanomic unit (OTUs), which was remarkably host-specific among locations. Wider dissimilarity in AMF communities was observed in small and disturbed (SD) sites compared to large and undisturbed (LU) sites, a trait that may be attributed to relaxed environmental filtering and facilitated AMF dispersal/immigration in SD sites from surrounding habitats. Overall, our results indicate that the assembly of root-colonizing AMF communities in the eastern Mediterranean sand dunes is characterized by strong biotic filtering (host identity), suggesting that co-adaptation processes may be more pronounced than previously proposed, under extreme environmental conditions.

Elevated heavy metal concentrations in top soils of an Aegean island town (Greece): total and available forms, origin and distribution.

Elevated heavy metal concentrations in urban top soils are principal indicators of environmental pollution; however, relative data on the heavy metal status in soils of Greek island towns, that are regional administrative centers and popular tourist destinations, are missing. A survey was conducted to examine heavy metal concentrations in the urban soils of Ermoupolis, the capital of Syros island and of the prefecture of Cyclades complex in the Aegean Sea. Total (aqua-regia extracted) and available (DTPA extracted) concentrations of Cu, Pb, Zn, Ni, Cr, Sn and Fe were determined in top soil samples collected from green areas and open spaces of the town and in surface samples from inland reference soils of the island. Mean values for the aqua-regia extracted fraction of Cu, Pb and Zn were 117, 155 and 440 mg kg(-1) respectively, up to four times higher than the respective mean values of the reference soils. Enrichment factors (EFs) for these metals indicated high accumulation in the urban top soils and the available to total concentration ratio of Cu, Pb, Zn and Fe was higher for the urban compared to the reference soils, suggesting differences in metal sequestration, resulting in higher metal availability in the urban soils. GIS analysis was used to visualize the spatial distribution of EFs of the studied heavy metals. Factor Analysis and Cluster Analysis, applied to aqua-regia and DTPA data sets, adequately elucidated the origin of metals grouped under each factor or cluster.

The use of hydroxyl-radical-generating systems for the treatment of olive mill wastewaters.

Three hydroxyl-radical producing biomimetic systems, composed of CuII, hydrogen peroxide and pyridine, glucaric or succinic acid, were able to perform decolorization of olive mill wastewaters (OMW) >85 % within 3 d combined with a significant removal of total phenols (>75 %). The systems consisting of 50 mmol/L succinic acid, 5-10 mmol/L CuSO4 and 100 mmol/L H2O2 were the most effective at OMW treatment, and led to the reduction of phenol contents to <1 % along with high decolorization (>88 %) and acceptable values of chemical oxygen demand.

Exploring the potential of biobeds for the depuration of pesticide-contaminated wastewaters from the citrus production chain: laboratory, column and field studies.

The high wastewater volumes produced during citrus production at pre- and post-harvest level presents serious pesticide point-source pollution for groundwater bodies. Biobeds are used for preventing such point-source pollution occurring at farm level. We explored the potential of biobeds for the depuration of wastewaters produced through the citrus production chain following a lab-to-field experimentation. The dissipation of pesticides used pre- or post-harvest was studied in compost-based biomixtures, soil, and a straw-soil mixture. A biomixture of composted grape seeds and skins (GSS-1) showed the highest dissipation capacity. In subsequent column studies, GSS-1 restricted pesticides leaching even at the highest water load (462 Lm(-3)). Ortho-phenylphenol was the most mobile compound. Studies in an on-farm biobed filled with GSS-1 showed that pesticides were fully retained and partially or fully dissipated. Overall biobeds could be a valuable solution for the depuration of wastewaters produced at pre- and post-harvest level by citrus fruit industries.

Evaluation of white-rot fungi for detoxification and decolorization of effluents from the green olive debittering process.

Wastewater produced by the debittering process of green olives (GOW) is rich in polyphenolics and presents high chemical oxygen demand and alkalinity values. Eight white-rot fungi ( Abortiporus biennis, Dichomitus squalens, Inonotus hispidus, Irpex lacteus, Lentinus tigrinus, Panellus stipticus, Pleurotus ostreatus and Trametes hirsuta) were grown in GOW for 1 month and the reduction in total phenolics, the decolorization activity and the related enzyme activities were compared. Phenolics were efficiently reduced by P. ostreatus (52%) and A. biennis (55%), followed by P. stipticus (42%) and D. squalens (36%), but only P. ostreatus had high decolorization efficiency (49%). Laccase activity was the highest in all of the fungi, followed by manganese-independent peroxidase (MnIP). Substantial manganese peroxidase (MnP) activity was observed only in GOW treated with P. ostreatus and A. biennis, whereas lignin peroxidase (LiP) and veratryl alcohol oxidase (VAOx) activities were not detected. Early measurements of laccase activity were highly correlated ( r(2)=0.91) with the final reduction of total phenolics and could serve as an early indicator of the potential of white-rot fungi to efficiently reduce the amount of total phenolics in GOW. The presence of MnP was, however, required to achieve efficient decolorization. Phytotoxicity of GOW treated with a selected P. ostreatus strain did not decline despite large reductions of the phenolic content (76%). Similarly, in GOW treated with purified laccase from Polyporus pensitius, a reduction in total phenolics which exceeded 50% was achieved; however, it was not accompanied by a decline in phytotoxicity. These results are probably related to the formation of phenoxy radicals and quinonoids, which re-polymerize in the absence of VAOx but do not lead to polymer precipitation in the treated GOW.

Adaptation and population dynamics of Azotobacter vinelandii during aerobic biological treatment of olive-mill wastewater.

Olive-mill wastewater (OMW) has a high organic and polyphenol content and is resistant to biodegradation. Its disposal leads to a major environmental pollution problem in the Mediterranean basin. The detoxification of OMW following inoculation with Azotobacter vinelandii (strain A) was performed for two successive 5-day-period cycles in an aerobic, biowheel-type reactor, under non-sterile conditions. The phytotoxicity of the processed product was reduced by over 90% at the end of both cycles. To exclusively monitor the A. vinelandii population in the reactor a most probable number-PCR approach was employed and applied daily to serial dilutions of total DNA extracted from reactor samples. PCR sensitivity was independent of the presence of OMW or non-target DNA. The A. vinelandii population dynamics were successfully monitored, showing an initial adaptation period, followed by a sharp population maximum on the fourth day of both cycles (1.6x10(8) and 9.6x10(7) cells ml(-1) respectively), after a major phytotoxicity decline. N(2) fixation rates were estimated using the acetylene reduction assay and reached a peak during the first 1-2 days of each cycle (36 and 29 nmol C(2)H(2) ml(-1) h(-1) respectively). The data are consistent with an initial physiological adaptation phase, where the presence of phenolic compounds limits A. vinelandii growth but stimulates N(2) fixation, followed by a rapid growth phase as phytotoxicity declines.