Manganese tolerance in Verbascum olympicum Boiss. affecting elemental uptake and distribution: changes in nicotinic acid levels under stress conditions.

A multielemental determination methodology in conjunction with an organic acid analysis that were supplemented with other stress parameters and an ultrastructural analysis used herein to study Verbascum olympicum Boiss. (Scrophulariaceae) under Mn stress. Uptake and accumulation characteristics of B, Cu, Fe, Mn, Mo, and Zn were evaluated in 8-week-old seedlings grown in Hoagland's nutrient solution and exposed to 5 (CK), 50, and 200 µM MnSO4 for 7 days. Hydrogen peroxide levels were determined to evaluate oxidative stress, and changes in compatible substance levels (total phenolic contents, glutathione and glutathione disulfide levels) were determined to assess antioxidant defense mechanisms. The distribution of manganese on the root surface was characterized by scanning electron microscopy images and energy-dispersive X-ray spectroscopy analysis. The levels of nicotinic acid, which is involved in nicotinamide adenine dinucleotide biosynthesis, were determined in roots and leaves to assess tolerance mechanisms. V. olympicum exhibited the ability to cope with oxidative stress originating from excessive Mn, while increased Mn concentrations were observed in both roots and leaves. The translocation factor of B was the most affected among other studied elements under the experimental conditions. Total nicotinic acid levels exhibited a trend of reduction in the roots and leaves, which could be attributed to the appropriate metabolic progress associated with oxidative stress based on the nicotinamide adenine dinucleotide cycle that may reach glutathione in response to manganese stress during plant growth.

Elemental Composition of Plant Species from an Abandoned Tungsten Mining Area: Are They Useful for Biogeochemical Exploration and/or Phytoremediation Purposes?

We aimed to evaluate the elemental (W, Mo, Zn, Fe, Cu, Co, Bi, Mn, Cd, Cr, As) composition of some plant species spread around the abandoned tungsten mining area of Uludag Mountain. The plant species tested were Anthemis cretica and Trisetum flavescens which are grown in this area and they are pioneer species on these contaminated sites. W levels in soils were found up to 1378.6 ± 672.3 mg/kg dry weight in contaminated areas. The leaf W contents of the selected plant species were found 41.1 ± 24.4 and 31.1 ± 15.5 mg/kg dry weight for A. cretica and T. flavescens, respectively. Our results indicate that the elemental composition of species changed by the increased tungsten and some element concentrations in soil without detrimental effect. So, these species can be useful tungsten removal and some elements from contaminated sites.

Elemental composition of Marrubium astracanicum Jacq. growing in tungsten-contaminated sites.

This study evaluates the elemental (W, Mo, Zn, Fe, Cu, Cd, Mn, Pb, Cr, Co, B, and Bi) composition of Marrubium astracanicum Jacq. (Lamiaceae), around the abandoned tungsten mine on Uludag Mountain, Turkey, to determine if it is an appropriate candidate for phytomonitoring and/or phytoremediation purposes. Three sample sites were selected around the mine for soil and plant sampling. Two sites approximately 500 m from the mine were assumed to be unpolluted sites. The other site was selected from a waste removal pool (WRP) and was assumed to be a polluted site. The soil and different organs (roots, leaves, and flowers) of plant samples were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) to determine the elemental content. The classic open wet digestion procedure was applied to the samples with 5 mL HNO3 and 3 mL H2O2 in a borosilicate glass vessel for the roots, leaves, and the flowers of the plants. Kjeldahl digestion was used for the soil samples. The W, Zn, Fe, Cu, Cd, Mn, Pb, B, and Bi contents were found to be higher in the soil samples from the waste removal pools compared with the samples from the unpolluted sites. We also found that the elemental composition of M. astracanicum has generally been increased by the activity of the tungsten mine, and there were significant correlations between the elemental contents of the soil samples and plant parts, except for Mo and Cr. The high level of many elements in the soil samples indicates the presence of contamination related to tungsten-mining activity on Uludag Mountain. Assessing the elemental contents of M. astracanicum, we can suggest this species as a candidate for phytoremediation purposes of W-contaminated sites due to its high W-accumulation capacity.

Evaluation of genotoxic and apoptotic potential of Hypericum adenotrichum Spach. in vitro.

Hypericum adenotrichum Spach. is an endemic plant from Turkey that is also used in folk medicine. In this study, following analyses of its chemical composition, the genotoxic/antigenotoxic effects of the methanol extract of H. adenotrichum in human lymphocyte culture were investigated using in vitro sister chromatid exchange, micronucleus and comet assays. In addition, the anti-growth effect of the extract was investigated in human breast cancer cell lines (MCF-7 and MDA-MB-231) using MTT and ATP viability assays. The mode of cell death was determined using fluorescence microscopy and biochemical methods. We found that the H. adenotrichum extract demonstrated cytotoxic and genotoxic effects in a cell type-dependent manner. At selected doses (125-500 µg/ml), the H. adenotrichum extract exhibited significant genotoxic activity in human lymphocytes, whereas it showed anti-growth effects on cancer cell lines between 0.2 and 100 µg/ml concentrations. The mode of cell death in cancer cells was shown to be apoptosis due to the presence of pyknotic nuclei, the cleavage of poly-(ADP-ribose) polymerase (PARP) and/or the activation of caspase-3. These results suggest that H. adenotrichum might show both cytotoxic and genotoxic effects depending on the cell type. This should be taken into account in its use for therapeutic purposes.

Ni-induced Changes in Nitrate Assimilation and Antioxidant Metabolism of Verbascum olympicum Boiss.: Could the Plant be Useful for Phytoremediation or/and Restoration Purposes?

Verbascum olympicum Boiss. (Scrophulariaceae) were studied as a candidate plant for remediating the Ni polluted soils. The metabolic responses, such as nitrate assimilation (nitrate reductase and glutamine synthetase activity) and antioxidant system activity [superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activity], of this species exposed to nickel in Hoagland's nutrient medium were investigated as remediation performance parameters. The accumulation of nickel and the variations in the content of some elements (B, Cu, Fe, Mg, Mn, Mo and Zn) and some growth parameters, such as the water content, biomass production, and contents of chlorophyll and soluble protein, were also examined. The accumulation of Ni in both the roots and leaves varied depending on the exposure times and doses. Increased oxidative stress was suggested by the increases in the activities of SOD, CAT and APX. Although some element contents were inhibited by Ni treatments, these inhibitory effects was decreased depending on the time, and even these elements are accumulated in roots. These results are the novelties in the use of this species in biotechnology.

Nitrogen mineralization in a high altitude ecosystem in the mediterranean phytogeographical region of Turkey.

Interrelations exist in the terrestrial ecosystems between the plant type and characteristics of nutrient uptake. Annual net nitrogen mineralization in soils of different plant communities in the high altitude zone of Spil mountain located in the Mediterranean phytogeographical region of Turkey was investigated throughout one year by field incubation method. Seasonal fluctuations resulting from field incubation were markedly higher in autumn and spring than summer. These are mainly associated with the changes in soil moisture being at minimum in the Mediterranean summer. A significant correlation was developed between the net Nitrate (kg NO3(-)-N ha week(-1)) production and soil water content (p<0.05; r = 0.316 in soil of 0-5 cm; r = 0.312 in soil of 5-15 cm). The results showed that the annual productivity of nitrogen mineralization shows different values depending on communities. Annual net ammonium (NH4(+)-N) production in the soils of each community was negatively estimated. However annual net nitrate (NO3(-)-N) production (0-15 cm) was higher in grassland (27.8 kg ha y(-1)) and shrub (25.0 kg ha y(-1)) than forest (12.4 kg ha y(-1)) community. While annual net N(min) values were close to each other in grassland (14.5 kg ha y(-1)) and shrub (14.1 kg ha y(-1)), but negative in forest community (-3.6 kg ha y(-1)). The reasons for these differences are discussed.

Nutrient status in soil of Ski runs in the sub-alpine belt of Uludag Mountain, Bursa, Turkey.

Large areas of land are disturbed in sensitive bio-diverse mountain environments by Skiruns. Restoration of vegetation on such disturbed mountain sites may be hampered by soil degradation but the severity and nature of the constraints is not well understood. This study was designed to compare the water holding and nutritional status of soil in three Ski runs which had different construction dates and disturbance levels, and the adjacent undisturbed site in the Abies bommuelleriana forest community in the sub-alpine belt of Uludag Mountain (Bithynian Olympus). The values of soil parameters were depressed in proportion to the disturbance level. Water holding capacity (WHC), total nitrogen (N), organic carbon (C) and calcium (Ca2+), magnesium (Mg2+) and potassium (K+) contents (mg kg(-1) dry weight) of soils in the Ski run which had the highest disturbance level were lower than that of the undisturbed adjacent sites. However the results indicated that the soil parameters were less degraded when secondary vegetation was growing on the disturbed areas.

Verbascum bombyciferum Boiss. (Scrophulariaceae) as possible bio-indicator for the assessment of heavy metals in the environment of Bursa, Turkey.

In this study, we determined the heavy metal content (Cd(2+), Cr(3+), Cu(2+), Fe(3+), Ni(2+), Pb(2+), and Zn(2+)) in the soil surrounding the roots and different organs of Verbascum bombyciferum Boiss. (Scrophulariaceae), which is endemic to Uludag Mountain, Bursa, Turkey. Plant samples were collected from roadsides, and heavy metal accumulation capabilities were tested. This is one of the pioneer species of ruderal plant communities on roadsides, building sites, rubbish dumps, etc. Different organs of plant samples (roots, stems, leaves, and flowers) and their soils were analyzed by inductively couple plasma optical emission spectroscopy for their heavy metal contents. Some of the analyzed heavy metals (Cd(2+), Cr(3+), Pb(2+), and Zn(2+)) were usually increased depending on the traffic in the sample sites, and this variation was also reflected in heavy metal content of plant samples. Our results show that this plant can be used as a bio-indicator species in the monitoring of increased Cd(2+), Cr(3+), Pb(2+), and Zn(2+) in the environment. We also concluded that V. bombyciferum have the capability of Cd(2+), Cr(3+), Cu(2+), Ni(2+), Pb(2+), and Zn(2+) accumulation.

Element content (Cu, Fe, Mn, Ni, Pb, and Zn) of the ruderal plant Verbascum olympicum Boiss. from East Mediterranean.

In this study, heavy metal content (Cu, Fe, Mn, Ni, Pb, Zn) was determined in soils and different organs of Verbascum olympicum Boiss. This species is endemic to Uludag and spreads on destroyed areas such as: roadsides, developed building areas, ski lift stations and sheep folds. Soils and different organs (roots, stems, leaves and flowers) of plant samples were analyzed using an atomic absorption spectrophotometer for determining the element content. Heavy metal contents in soils and different organs in this species were highly correlated (P < 0.05). However, the contribution of plant organs to the accumulation capacity varied according to the metal. These results suggest that this species may be useful as a bioindicator for heavy metals.

Germination responses to GA3 and stratification of threatened Festuca L. species from Eastern Mediterranean.

The seed germination characteristics of three threatened Festuca sp. [F. punctoria Sm., F. cyllenica Boiss. et Heldr. subsp. uluana Markgr.-Dannenb., F. paphlagonica (St.-Yves) Markgr.-Dannenb. subsp. paphlagonica] were investigated. These species are endemic and spread on alpine belt. The study was carried out with wet-cold and dry-cold stratification throughout 15 days, different doses of GA3 (50, 100 and 150 ppm) and hormone-stratification combined treatments, and non-treatment series. We found that the germination rates of three fescue seeds for various treatment series were different. The mean germination percentage of F. cyllenica was higher (80%) than that of F. punctoria and F. paphlagonica which were fairly low (50-60%). Germination rates increased by wet-stratification treatment in F. punctoria and also increased with 100 ppm GA3 application to the seeds of F. paphlagonica. When taken into consideration the germination percentages of all fescue species, the seeds of F. punctoria and F. paphlagonica can be dormant, but the seeds of F. cyllenica are non-dormant.

Protein mobilization and proteolytic enzyme activities during seed germination of broad bean (Vicia faba L.).

The protein mobilization from attached and detached seeds of Vicia faba L. cv. Eresen 87 (Fabaceae) was investigated. While the total soluble protein content decreased, the free amino acid content increased during the 7 days germination period. Among the three proteolytic enzymes, only endopeptidase activity was found to be affected by the removal of the embryonic axis. Leucine aminopeptidase activity was high at the beginning, then it decreased; carboxypeptidase activity reached the highest value at day 5. In order to examine the effects of plant growth regulators on detached cotyledons incubated with plant growth regulators [10(-4) M benzyladenine (BA), gibberellic acid (GA3), indole acetic acid (IAA) and 10(-5) M abscisic acid (ABA)], only benzyladenine was found promotive on protein mobilization. Our results suggest that the removal of the embryonic axis in seeds of Vicia faba L. cv. Eresen 87 decreases protein mobilization and endopeptidase activity.

Composition and antimicrobial activity of the essential oil of Achillea multifida.

The composition of the water-distilled essential oil of Achillea multifida (DC.) Boiss. (Compositae) was analysed by GC and GC/MS. Fifty-eight compounds were identified representing 93.9 % of the total oil. alpha-Thujone (60.9 %), beta-thujone (9.1 %), sabinene (4.1 %) and camphor (3.7 %) were characterised as the main constituents. The essential oil was tested for its antimicrobial activity using a micro-dilution assay resulting in the inhibition (MIC: 62.5-250 microg/ml) of human pathogenic bacteria and yeast.