Sławomir Dresler

Morphological, physiological, and genetic variation between metallicolous and nonmetallicolous populations of Dianthus carthusianorum

Waste deposits produced by metal mining and smelting activities provide extremely difficult habitats for plant colonization and growth. Therefore, plants spontaneously colonizing such areas represent a very interesting system for studying evolution of plant adaptation and population differentiation between contaminated and noncontaminated environments. In this study, two populations of Dianthus carthusianorum, one originating from Zn-Pb waste deposit (a metallicolous population, M) and the other from unpolluted soil (a nonmetallicolous population, NM), were analyzed in respect of their morphological and physiological traits as well as genetic markers. It was found that the plants inhabiting the waste heap differed significantly from the NM plants in terms of leaf size and shape, and these differences were persistent between the first generation of the plants of both populations cultivated under uniform, controlled laboratory conditions. In contrast with the evident morphological differences, no significant differentiation between the populations regarding the physiological traits measured (accumulation of proline, anthocyanins, chlorophyll, carotenoids) was found. These traits can be regarded as neither population specific nor stress markers. The genetic variability was analyzed using 17 random amplified polymorphic DNA (RAPD) and four inter simple sequence repeat (ISSR) markers; this proved that the differentiation between the M and NM populations exists also at the genetic level. Analysis of molecular variance (AMOVA) showed that 24% of the total genetic diversity resided among populations, while 76% - within the populations. However, no significant differences in intrapopulation genetic diversity (Hj) between the M and NM populations of D. carthusianorum was found, which contradicts the theory that acquisition of adaptation mechanisms to adverse, isolated growth habitats is related to reduction in genetic diversity. Distinct genetic differences between the two populations in combination with evident morphological variation support the proposal to regard the M population of D. carthusianorum as a separate calamine ecotype.

The effect of silicon on maize growth under cadmium stress

The effects of silicon (Si) supply (0, 0.1, 0.5, 1.5, 3.0, and 5.0 mM Si) on maize seedling growth, Si and Cd accumulation, and thiol peptide synthesis under Cd stress conditions were studied. The addition of Si to the growth medium resulted in the significantly higher Si accumulation in plant tissues. The average values of growth parameters (root and shoot fresh weights and root net elongation rates) showed a beneficial role of Si on growth of non-Cd-treated plants, while there was no evidence that silicon mitigated Cd toxicity in maize seedlings. Cadmium exposure depressed plant growth and induced phytochelatin (PC) synthesis. The accumulation of Cd and PCs in roots significantly decreased with increasing Si concentrations in the nutrient solution; however, their accumulation in shoots was not changed in the presence of Si.

Effect of cadmium on selected physiological and morphological parameters in metallicolous and non-metallicolous populations of Echium vulgare L.

Cadmium tolerance of three populations of Echium vulgare L., naturally occurring on two Zn-Pb waste deposits (metallicolous populations M1, M2) and on an uncontaminated site (non-metallicolous population, NM) was investigated. The plants were cultivated in hydroponics at 0, 5, 15, 30, or 50μM Cd for 14 days. Although Cd reduced the content of photosynthetic pigments indifferently in the three populations, plant growth parameters and root viability analyses confirmed different Cd tolerances decreasing in the order M1>M2>NM in the populations studied. Organic acids (tartrate, malate, citrate, succinate) were not responsible for the elevated Cd tolerance of the metallicolous populations, although malate and citrate might participate in Cd detoxification in the roots of the M1 and M2. Phytochelatin concentrations were higher in the roots of M1 and M2 populations of E. vulgare, suggesting their role in Cd detoxification and different Cd tolerances.

Determination of Shikonin and Rosmarinic Acid in Echium vulgare L. and Echium russicum J.F. Gmel. by Capillary Electrophoresis

A simple and rapid capillary zone electrophoresis (CZE) procedure for determination of shikonin and rosmarinic acid in roots of Echium vulgare L. and Echium russicum J.F. Gmel. is described. The separation was achieved in less than 4 min. The analysis was performed in fused silica capillary (i.d. 75 µm, 48.5 cm total length) using 50 mM borate, at pH 9.5 as the run buffer with applied voltage of 30 kV and constant temperature (27 °C). The method ensures good linearity and repeatability with high correlation coefficients (0.994 and 0.998 for shikonin and rosmarinic acid, respectively). The compounds were detected by UV absorption at 218 (shikonin) and 202 nm (rosmarinic acid). The limits of detection (LOD) for shikonin and rosmarinic acid were 0.603 ppm and 0.270 ppm, respectively. The method was used for estimation of both components in Echium plants. We found out that the content of shikonin and rosmarinic acid were 4.4–4.8 mg/kg air-dry material and 20.0–54.2 mg/kg air-dry material, respectively, in the Echium species analyzed.

Comparison of some secondary metabolite content in the seventeen species of the Boraginaceae family

Abstract Context: The Boraginaceae family comprises plants that have important therapeutic and cosmetic applications. Their pharmacological effect is related to the presence of naphthaquinones, flavonoids, terpenoids, phenols, or purine derivative – allantoin. Objective: In the present study, comparison of some secondary metabolite content and phytochemical relationship between 17 species of the Boraginaceae family were analyzed. Materials and methods: High performance capillary electrophoresis (HPCE) was used to perform a chemometric analysis in the following Boraginaceae species: Anchusa azurea Mill., Anchusa undulata L., Borago officinalis L., Buglossoides purpurocaerulea (L.) I.M. Johnst., Cerinthe minor L., Cynoglossum creticum Mill, Echium italicum L., Echium russicum J.F. Gmel., Echium vulgare L., Lindelofia macrostyla (Bunge) Popov (syn. Lindelofia anchusoides (Lindl.) Lehm.), Lithospermum officinale L., Nonea lutea (Desr.) DC., Omphalodes verna Moench (syn. Cynoglossum omphaloides L.), Pulmonaria mollis Wulfen ex Hornem., Pulmonaria obscura Dumort., Symphytum cordatum Waldst. & Kit ex Willd., and Symphytum officinale L. Results: Six active compounds in shoot extracts (allantoin, p-hydroxybenzoic acid, rutin, hydrocaffeic acid, rosmarinic acid, and chlorogenic acid) and four compounds in root extracts (allantoin, hydrocaffeic acid, rosmarinic acid, and shikonin) were identified. The presence and abundance of these compounds were used for the characterization of the species and for revealing their phytochemical similarity and differentiation. Discussion and conclusion: The present study provides the first comprehensive report of the extraction and quantification of several compounds in Boraginaceae species (some of them for the first time). Among the 17 species studied, species with potentially high pharmacological activity were recognized.

Does methyl jasmonate modify the oxidative stress response in Phaseolus coccineus treated with Cu

The contribution of methyl jasmonate (MJ) as a signal molecule able to take part in the defense mechanism against copper (Cu)-imposed oxidative stress was studied in the leaves and roots of runner bean (Phaseolus coccineus) plants. Roots of plants cultivated hydroponically were preincubated in MJ (10µM) for 1h or 24h and subsequently exposed to Cu (50µM) for 5h (short-term experiment) or 5 days (long-term experiment). Enzymatic (activity of superoxide dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX; guaiacol peroxidase, POX) and non-enzymatic (accumulation of malondialdehyde, MDA; homoglutathione, hGSH; proline; anthocyanins; low molecular weight organic acids, LMWOAs) responses were determined in the leaves and roots. The antioxidative defense mechanism was significantly activated after Cu supplementation. In most cases, activities of ROS (reactive oxygen species) scavenging enzymes like SOD, CAT, APX, POX, as well as MDA, hGSH and proline concentrations increased following Cu exposure. MJ showed a time-dependent effect on antioxidative enzymes activity. In the short-term experiment, MJ elevated CAT, APX and POX activities in the roots, and POX activity in the leaves of non-Cu-treated plants. In the long-term experiment, MJ not only decreased POX and partially CAT activity in the roots, but also increased the MDA level and partially CAT activity in the leaves of the control plants. In Cu-treated plants, MJ reduced APX, but elevated POX activity in the leaves after 5-h exposure. After 5-day-Cu treatment, MJ inhibited POX activity in the leaves and mainly reduced SOD and CAT activities in the roots. Moreover, in the long-term experiment, MJ reduced tartrate and pyruvate in the leaves of Cu-stressed plants, but mostly elevated tartrate and malate in the roots comparing with Cu alone treatment. MJ alone and under Cu excess did not alter accumulation of MDA, hGSH and proline comparing with Cu alone, but partially elevated anthocyanin concentration. The results indicated that MJ was both partially potent in modifying the antioxidative enzymes activity and metabolites accumulation in non-stress and Cu-stress conditions.

Biofortification of soy (Glycine max (L.) Merr.) with strontium ions.

Soy (Glycine max (L.) Merr.) is an annual plant cultivated worldwide mostly for food. Moreover, due to its pharmacological properties it is widely used in pharmacy for alleviating the symptoms of osteoporosis. The aim of the present study was to investigate the biofortification of soy treated with various concentrations of strontium. Soy was found to have a strong capacity to absorb Sr(2+) (bioconcentration factor higher than 1). A positive linear correlation (R(2) > 0.98) between the amount of strontium in the growth medium and its content in the plant was also observed. Moreover, at a concentration of 1.5 mM, strontium appeared to be nontoxic and even stimulated plant growth by approximately 19.4% and 22.6% of fresh weight for shoots and roots, respectively. Our research may be useful to obtain vegetable products or herbal preparations containing both phytoestrogens and strontium to prevent postmenopausal osteoporosis.

The Stimulatory Effect of Strontium Ions on Phytoestrogens Content in Glycine max (L.) Merr

The amount of secondary metabolites in plants can be enhanced or reduced by various external factors. In this study, the effect of strontium ions on the production of phytoestrogens in soybeans was investigated. The plants were treated with Hoagland’s solution, modified with Sr2+ with concentrations ranging from 0.5 to 3.0 mM, and were grown for 14 days in hydroponic cultivation. After harvest, soybean plants were separated into roots and shoots, dried, and pulverized. The plant material was extracted with methanol and hydrolyzed. Phytoestrogens were quantified by HPLC. The significant increase in the concentration of the compounds of interest was observed for all tested concentrations of strontium ions when compared to control. Sr2+ at a concentration of 2 mM was the strongest elicitor, and the amount of phytoestrogens in plant increased ca. 2.70, 1.92, 3.77 and 2.88-fold, for daidzein, coumestrol, genistein and formononetin, respectively. Moreover, no cytotoxic effects were observed in HepG2 liver cell models after treatment with extracts from 2 mM Sr2+-stressed soybean plants when compared to extracts from non-stressed plants. Our results indicate that the addition of strontium ions to the culture media may be used to functionalize soybean plants with enhanced phytoestrogen content.

Physiological mechanisms of adaptation of Dianthus carthusianorum L. to growth on a Zn-Pb waste deposit - the case of chronic multi-metal and acute Zn stress

AimsThis study investigates the response of metallicolous (M) and nonmetallicolous (NM) ecotypes of Dianthus carthusianorum L. to chronic multi-metal and acute Zn stress.MethodsPlants were cultivated on the Zn-Pb waste heap substrate and under Zn excess in hydroponics. Growth parameters as well as accumulation of organic acids and thiol peptides were determined as a function of metal accumulation.ResultsWhen grown on the metalliferous substrate, the M plants showed less phytotoxicity symptoms, lower foliar metal (Zn, Pb, Cd) accumulation, higher malate and citrate but lower glutathione content than the NM plants. When exposed to Zn excess in hydroponics, the M ecotype was also more tolerant but accumulated more Zn in comparison with the NM ecotype, accompanied by greater malate and citrate concentrations in the shoots, which were however not affected by increasing Zn doses. No phytochelatins were detected under any experimental conditions.ConclusionsBoth constitutive and adaptive tolerance was found in D. carthusianorum. Under chronic metal stress, enhanced tolerance results from restricted metal uptake to the shoots and probably from detoxification by organic acids; however, under acute Zn stress it is not related to diminished metal uptake or organic acids. Glutathione and phytochelatins are not implicated in adaptive metal tolerance.

Rapid Determination of Coumarins in Plants by Capillary Electrophoresis

The goal of this study was to optimize the parameters for capillary electrophoresis to separate and determine aesculin, aesculetin, umbelliferone, and dihydrocoumarin in plant materials. A simple and rapid capillary zone electrophoresis method is reported for this separation that required less than nine minutes. A bare fused silica capillary was employed using 20 mM borax in 5% methanol at pH 10.1 as the background electrolyte with an applied voltage of 30 kV at 27°C. Good linearity and reproducibility were obtained with high correlation coefficients. The analytes were determined by molecular absorption at 194 and 206 nm. Coumarins were determined in Aesculus hippocastanum L., Cichorium intybus L., Melilotus officinalis L., and Juniperus communis L. “Pendula.” The concentrations of aesculin and aesculetin were 3.07 and 6.31 mg g−1 in Aesculus hippocastanum. In Cichorium intybus, the aesculetin concentration was 2.42 mg g−1. The dihydrocoumarin concentration was 0.54 mg g−1 in Melilotus officinalis, and the concentration of umbelliferone was 0.58 mg g−1 in Juniperus communis “Pendula.”