Milan Pavlík

The effect of arsenic contamination on amino acids metabolism in Spinacia oleracea L.

Changes of amino acid concentrations (proline, glutamate, asparagine, aspartate, alanine) and glutamate kinase activity (GKA) in plants under arsenic chronic stress reported here reveal their role in plant arsenic stress adaptation. Results of the pot experiment confirmed the toxic effect of arsenic at tested levels (As1=25 mg As kg(-1) soil, As2=50 mg As kg(-1) soil, As3=75 mg As kg(-1) soil) for spinach. Growing available arsenic contents in soil were associated with the strong inhibition of above-ground biomass and with the enhancement of As plant content. The changes of glutamate, asparagine, aspartate and proline levels in the plants showed strong linear dependences on arsenic concentration in plants (R2=0.60-0.90). Compared to the untreated control, concentrations of free proline and aspartate of As3 treatment were enhanced up to 381% and 162%, respectively. The significant changes of glutamate were observed on As2 and As3 treatments (increased level up to 188, i.e. 617%). Arsenic in plants was shown to be an inhibitor of glutamase kinase activity (R2=0.91). Inhibition of GKA resulted in an increase in the content of glutamate that is used in synthesis of phytochelatins in plant cells. Concentration of alanine did not have a confirmed linear dependence on arsenic concentration in plant (R2=0.05). The changes of its concentrations could be affected by changes of pH in plant cell or induction of alanine aminotransferase by hypoxia.

Trace elements present in airborne particulate matter—Stressors of plant metabolism

Changes of amino acid concentrations (glutamic acid, glutamine, asparagine, aspartate, proline, tryptophan, alanine, glycine, valine and serine), gas-exchange parameters (net photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO(2) concentration) and nitrate levels in Lactuca serriola L. under airborne particulate matter (PM) contamination reported here reveal their role in plant chronic stress adaptation. Results of the pot experiment confirmed the toxic effect of trace elements present in PM for lettuce. PM applied to soil or on the lettuce leaves were associated with the strong inhibition of above-ground biomass and with the enhancement of plant trace element contents. The significant changes of amino acid levels and leaf gas-exchange parameters of the plants showed strong linear dependences on PM contamination (R(2)=0.60-0.99). PM application on leaves intensified toxic effect of trace elements (As, Pb, Cr and Cd) originating from PM by shading of the leaf surface. The plant accumulation of nitrate nitrogen after PM contamination confirmed to block nitrate assimilation.

The long-term effect of zinc soil contamination on selected free amino acids playing an important role in plant adaptation to stress and senescence

Increased endogenous plant cytokinin (CK) content through transformation with an isopentyl transferase (ipt) gene has been associated with improved plant stress tolerance. The objective of this study is to determine amino acid changes associated with elevated CK production in ipt transgenic tobacco (Nicotiana tabacum L., cv. Wisconsin 38). Nontransformed (WT) and transformed tobacco plants with ipt gene controlled by senescence-activated promoter (SAG) were exposed to zinc soil contamination (tested levels Zn1=250, Zn2=500, Zn3=750 mg kg(-1) soil). The Zn effect on plant stress metabolism resulted in changes in levels of selected free amino acids playing an important role in adaptation to stress and plant senescence (alanine, leucine, proline, methionine and γ-aminobutyrate) and differed for transformed and nontransformed tobacco plants. Analyses of amino acids confirmed that SAG tobacco plants had improved zinc tolerance compared with the WT plants. The enhanced Zn tolerance of SAG plants was associated with the maintenance of accumulation of proline, methionine and γ-aminobutyrate. The concentrations of leucine and alanine did not show significant differences between plant lines.

High-performance liquid chromatographic analysis and separation of N-feruloylserotonin isomers.

The N-feruloylserotonin containing fraction was isolated from seeds of Leuzea carthamoides (Willd.) DC by solvent extraction followed by column chromatography on silica gel or on Sephadex LH-20. Nuclear magnetic resonance spectroscopic analysis of the isolated fraction showed the presence of four structurally related compounds. These compounds were identified as four isomers of N-feruloylserotonin: N-(Z)-feruloylserotonin, N-(Z)-isoferuloylserotonin, N-(E)-feruloylserotonin and N-(E)-isoferuloylserotonin. They were analyzed by HPLC on Separon SGX C18, Separon SGX and Separon SGX phenyl, using various mobile phases. Separon SGX phenyl phase was found the most efficient for a rapid analysis and for the final separation of the N-feruloylserotonin isomers.

Nitrogen metabolism and gas exchange parameters associated with zinc stress in tobacco expressing an ipt gene for cytokinin synthesis

Increased endogenous plant cytokinin (CK) content through transformation with an isopentyl transferase (ipt) gene has been associated with improved plant stress tolerance. The impact of zinc (tested levels Zn1=250, Zn2=500, Zn3=750mgkg(-1)soil) on gas exchange parameters (net photosynthetic rate, transpiration rate, stomatal conductance, intercellular CO2 concentration) and nitrogen utilization by plants resulted in changes of free amino acid concentrations (glutamic acid, glutamine, asparagine, aspartate, glycine, serine, cystein) and differed for transformed and non-transformed tobacco plants. For pot experiments, tobacco plants (Nicotiana tabacum L., cv. Wisconsin 38) transformed with a construct consisting of SAG12 promoter fused with the ipt gene for cytokinin synthesis (SAG plants) and its wild type (WT plants as a control) were used. Physiological analyses confirmed that SAG plants had improved zinc tolerance compared with the WT plants. The enhanced Zn tolerance of SAG plants was associated with the maintenance of accumulation of amino acids and with lower declines of photosynthetic and transpiration rates. In comparison to WT plants, SAG plants exposed to the highest Zn concentration accumulated lower concentrations of asparagine, which is a major metabolic product during senescence.

Monitoring Native Vegetation on a Dumpsite of PCB-Contaminated Soil

Composition of native vegetation on a polychlorinated biphenyls (PCB)-contaminated soil dumpsite at Lhenice, South Bohemia (Czech Republic), was determined and species variability in the accumulation of PCBs in plant biomass was investigated. Soil stripping contaminated by PCBs originated at a factory producing electrical transformers that mostly used the commercial PCB mixture Delor 103 and 106. The PCB content of soil in the most contaminated part of the dumpsite reached 153 mg kg−1 dry soil. Low diversity of plant species was found on the dumpsite. Results showed three grass species, Festuca arundinacea Schreb., Phalaroides arundinacea (L.) Rauschert., and Calamagrostis epigeios (L.) Roth., to be the major components of the vegetation and confirmed their high tolerance toward PCB contamination. The highest content of PCB in plant biomass—813.2 μg kg−1 dry biomass—was determined in Festuca aboveground biomass. For phytoextraction purposes especially, Festuca can be recommended due to its high biomass yield, but its bioconcentration factor was very low (0.006). Tripleurospermum maritimum (L.) Sch. Bip. and Cirsium arvense (L.) Scop. grew mainly at the margins of the most contaminated part of the dumpsite. The PCB content determined in their aboveground biomass—278.7 and 289.5 μg kg−1 dry biomass, respectively—was nonsignificantly lower compared to grass species Phalaroides and Calamagrostis. Salix (Salix viminalis L. and Salix caprea L.) was monitored among plant species composition at this site as a representative of woody species.

Chiral juvenoids derived from 2-substituted cyclohexanols

Abstract Chiral juvenoids 1 - 4d were prepared by a chemoenzymatic process consisting in employing microorganism-mediated biotransformations and/or enzyme-mediated transformation of convenient substrates, followed by a chemical transformation of the chiral intermediates into the chiral compounds targeted. Biological activity of the juvenoid stereoisomers 1d - 4d on the yellow mealworm ( Tenebrio molitor ) pupae was studied.

Long-term effects on food choice of land snail Arianta arbustorum mediated by petasin and furanopetasin, two sesquiterpenes from Petasites hybridus

Sesquiterpenes (STs) from the Senecioneae have been found to be potent snail repellents. We investigated the range of activity of the STs petasin, isopetasin, furanopetasin, kablicin, and cacalol, which were isolated from Petasites hybridus, P. kablikianus, and Adenostyles alpina. We found the petasin content of leaves of P. hybridus to lie within the range of deterrence of the isolated compound. Furthermore, leaf extracts containing petasin proved to be deterrent, and leaf discs with low petasin content were preferred over discs with higher petasin content. The cacalol-containing fraction of a leaf extract of A. alpina was not deterrent to the snails. When the snails had experience with the relevant ST one week before a choice test, their sensitivity towards petasine and furanopetasine increased whereas for the other ST it remained at the same level. We speculate that this sensitivity increase could be the result of a rapid long-term associative learning process, but there is also the possibility that these STs are directly interfering with the feeding motor program of the snails, thereby eliciting a direct neurophysiological sensitization reaction which prevents them from further feeding.

Sesquiterpenes from the Senecioneae and their effect on food choice of the specialised leaf beetles Oreina cacaliae, Oreina speciosissima and the generalist snail Arianta arbustorum

B e r n d H~igele 1 , Juraj H a r m a t h a 2, Mi l an Pav l fk 2 & Mar t i n e R o w e l l R a h i e r 3 1 Zoologisches Institut der Universitiit Basel, Rheinsprung 9, CH-4051 Basel, Switzerland 2 Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nrm. 2, 16610 Prague, Czech Republic 3 lnstitut de Zoologie, Universit~ de Neuch~tel, Rue Emile-Argand 11, CH-2007 Neuch6tel, Switzerland

Fatty acid profiles of ecotypes of hyperaccumulator Noccaea caerulescens growing under cadmium stress

Changes in the fatty acid (FAs) composition in response to the extent of Cd contamination of soils (0, 30, 60 and 90 mg Cd kg(-1)) differed between ecotypes of Noccaea caerulescens originating from France - Ganges, Slovenia - Mežica and Austria - Redlschlag. Mežica ecotype accumulated more Cd in aboveground biomass compared to Ganges and Redlschlag ecotypes. Hyperaccumulators contained saturated fatty acids (SFAs) rarely occurring in plants, as are cerotic (26:0), montanic (28:0), melissic (30:0) acids, and unusual unsaturated fatty acids (USFAs), as are 16:2, 16:3, 20:2 and 20:3. Typical USFAs occurring in the family Brassicaceae, such as erucic, oleic and arachidonic acids, were missing in tested plants. Our results clearly indicate a relationship between Cd accumulation and the FAs composition. The content of SFAs decreased and the content of USFAs increased in aboveground biomass of Ganges and Mežica ecotypes with increasing Cd concentration. Opposite trend of FAs content was determined in Redlschlag ecotype. Linoleic (18:2n-6), α-linolenic (18:3n-3) and palmitic (16:0) acids were found in all ecotypes. The results observed in N. caerulescens ecotypes, showed that mainly Mežica ecotype has an efficient defense strategies which can be related on changes in FAs composition, mainly in VLCFAs synthesis. The most significant effect of ecotype on FAs composition was confirmed using multivariate analysis of variance.