Glutathione and phytochelatin contents in tomato plants exposed to cadmium

The effect of cadmium on growth and contents of glutathione (GSH) and phytochelatins (PCs) were investigated in roots and leaves of tomato plants (Lycopersicon esculentum Mill. cv. 63/5 F1). The accumulation of Cd increased with external Cd concentrations and was considerably higher in roots than in leaves. Dry mass production decreased under Cd treatment especially in leaves. In both roots and leaves, exposure to Cd caused an appreciable decline in GSH contents and increase in PCs synthesis proportional to Cd concentrations in the growth medium. At the same Cd concentration, PCs production was higher in roots than in leaves. The implication of glutathione in PC synthesis was strongly suggested by the use of buthionine sulfoximine (BSO). The major fraction of Cd accumulated by tomato roots was in the form of a Cd-PCs complex.     

Effects of phosphorus and chilling under low irradiance on photosynthesis and growth of tomato plants

To determine the effects of phosphorus nutrition on chilling tolerance of photosynthetic apparatus, tomato (Lycopersicon esculentum Mill. cv. Kenfengxin 2002) plants were raised under different P contents and subjected to 7 d of chilling at 9/7 °C. After chilling (2 h or 7 d) plant growth, P content in tissue, gas exchange and chlorophyll fluorescence were measured. Decreasing P concentration [P] in the nutrient solution markedly reduced plant growth and the chilled plants exhibiting higher optimum [P] than the unchilled plants. Decreasing [P] significantly decreased light saturated net photosynthetic rate (P_Nsat), maximum carboxylation velocity of Rubisco (V_cmax), maximum potential rate of electron transport contributed to Rubisco regeneration (J_max), quantum efficiency of photosystem (PS) 2 (ΠPS2) and O₂ sensitivity of P_Nsat (PS_O2) and this trend was especially apparent in chilled plants.     

Effect of boron supply on nitrate concentration and its reduction in roots and leaves of tobacco plants

Shoot and root mass of tobacco plants treated with only 0.05 μM boron was decreased by 25 and 50 %, respectively, when compared to plants sufficiently supplied with B (2 and 5 μM). Leaf B content of 0.05 μM B-treated plants decreased (about 80–90 %) when compared to 2 μM B treated plants; this drop of B content were not as marked (about 25–45 %) in roots. Leaf and root nitrate contents in B-deficient plants were 45–60 % and 35–45 % lower, respectively, than those from 2 and 5 μM B treated plants. It is suggested that B deficiency might decrease nitrate uptake rather than nitrate reductase activity in tobacco plants.     

Contribution of putrescine degradation to proline accumulation in soybean leaves under salinity

Proline accumulation was studied in the leaves of Glycine max (L.) Merr. subjected to salt stress in the presence of aminoguanidine (AG, a specific inhibitor of diamine oxidase, DAO) and exogenous putrescine (Put). Both DAO activity and proline content were increased while endogenous Put content was decreased in soybean leaves under 50 to 150 mM NaCl. There was a negative correlation between proline accumulation and endogenous Put content. The addition of AG during NaCl stress inhibited DAO activity, caused Put accumulation and a 15 to 20 % decrease in proline content. Application of 1 mM Put to NaCl solution markedly increased proline content. The promotive effect of Put application could be alleviated by the treatment with Put plus AG. Moreover an application of AG had no effect on proline accumulation in soybean seedlings grown under normal condition. These results indicate that the quantitative contribution of Put degradation to proline formation is 15 to 20 %.     

Expression of tea cytosolic glutamine synthetase is tissue specific and induced by cadmium and salt stress

Glutamine synthetase (GS) showed highest expression and activity in bud (youngest topmost leaf) of Camellia sinensis, lower in older leaves, while lowest activity in stem and roots. GS expression and activity was increased by ammonium and nitrate and also by cadmium and salt stress but decreased by copper, aluminum, drought, cold and heat stress.     

In vivo activity of recombinant human lewis fucosyltransferase III in leaves of Nicotiana tabacum L.

Fucosylation in plants occurs in glycoproteins and polysaccharides but the function of fucosylation is largely unknown. We aimed to analyze the effects of over-expression of human fucosyltransferase III (hFucT III) on in vivo N-glycan accumulation in tobacco plant leaves and focused on comparing the amount of Lewis a (Le^a)-epitope accumulation in transgenic and in wild-type plants. Fucosyltransferase assays, Western blot and mass spectrometry were used to identify, quantify and analyse Le^a N-glycans. We found that constitutive overexpression of hFucT III activity had no effect on Le^a complex type N-glycans accumulation. Our results suggest that tobacco recombinant hFucT III acts more as a hydrolase than as a transferase.     

Physiological responses of <Emphasis Type="Italic">Lupinus luteus</Emphasis> to different copper concentrations

Yellow lupin (Lupinus luteus L.) plants were grown in hydroponic solution for 15 d under different copper concentrations (0.1, 0.5, 1.0, 10, 25 and 50 μM). With increasing Cu concentration total biomass was not affected, leaf area slightly decreased, while chlorophyll content decreased considerably. Cu content increased significantly both in roots and in leaves, but the contents of other ions were only slightly affected at the highest Cu concentration (Mn content decreased both in roots and in leaves, P content decreased only in leaves and Zn content increased in roots). Superoxide dismutase (SOD) activity increased up to day 7 after copper application. Peroxidase (GPOD) and polyphenol oxidase (PPO) activities also increased, while catalase (CAT) activity remained constant.     

A novel cadmium induced protein in wheat: characterization and localization in root tissue

A 51-kDa soluble protein was over-expressed in wheat (Triticum aestivum) seedlings by the treatment of seeds before germination with 50 μM CdCl₂ for 48 h and subsequently washed off Cd²⁺. This protein designated as Cd stress associated protein (CSAP), was purified. Polyclonal antibody was raised against CSAP for localizing the protein in root tissue of treated and control seedlings. It was observed that CSAP was located below the plasma membrane and outer periphery of the tonoplast. This unique type of organized localization of CSAP is suggestive of defensive role against metal phytotoxicity. N-terminal analysis of CSAP and expressed sequence tags (EST) database search of wheat sequences suggests that this protein has not been reported earlier in higher plants.     

The differences in cell wall composition in leaves and regenerating protoplasts of Beta vulgaris and Nicotiana tabacum

Cell wall composition in leaves and regenerating leaf-derived protoplasts was studied in Beta vulgaris L. and Nicotiana tabacum L. Several oligosaccharides that characterize arabinogalactan proteins (AGPs) and pectins were visualized in situ by a series of immunocytochemical reactions. The most conspicuous difference between the two species involved the expression of AGP epitopes that bind LM2 and MAC207 in only sugar beet cells; both epitopes being characterized by the presence of glucuronic acid (GlcA). Moreover, B. vulgaris leaves and protoplast-derived cells showed lower content of pectin side-chains bearing (1→4)-β-D-galactose residues as compared to N. tabacum.     

Irreversible changes in barley leaf chlorophyll fluorescence detected by the fluorescence temperature curve in a linear heating/cooling regime

The chlorophyll fluorescence (F) temperature curves in a linear time-temperature heating/cooling regime were used to study heat-induced irreversible F changes in primary green leaves of spring barley (Hordeum vulgare L. cv. Akcent). The leaf segments were heated in a stirred water bath at heating rates of 0.0083, 0.0166, 0.0333, and 0.0500 °C s⁻¹ from room temperature up to maximal temperature T _m and then linearly cooled to 35 °C at the same rate. The F intensity was measured by a pulse-modulated technique. The results support the existence of the two critical temperatures of irreversible F changes postulated earlier, at 45–48 and 53–55 °C. The critical temperatures are slightly dependent on the heating rate. Two types of parameters were used to characterize the irreversibility of the F changes: the coefficient of irreversibility μ defined as the ratio of F intensity at 35 °C at the starting/ending parts of the cycle and the slopes of tangents of linear parts of the F temperature curve. The dependence of μ on T _m revealed a maximum, which moved from 54 to 61 °C with the increasing heating/cooling rate v from 0.0083 to 0.0500 °C s⁻¹, showing two basic phases of the irreversible changes. The Arrhenius and Eyring approaches were applied to calculate the activation energies of the initial increase in μ. The values varied between 30 and 50 kJ mol⁻¹ and decreased slightly with the increasing heating rate.     

Agrobacterium-mediated genetic transformation of plants: The role of host

Agrobacterium-mediated genetic transformation is the most widely used technology to obtain overexpression of recombinant proteins in plants. Molecular events that occur within Agrobacterium during interactions with host plants have been studied extensively, and now we have a reasonable understanding the key factors involved in the regulation of T-DNA nuclear import and genomic integration. By contrast, very little is known about the events that take place in the host cells during genetic transformation by Agrobacterium. Here, we describe the plant-related factors including genotype, genes, proteins, competency of target tissues and phenolic compounds that participate in Agrobacterium-mediated genetic transformation and discuss their possible roles in this process. Because Agrobacterium probably adapts existing cellular processes for its life cycle, identifying the processes in host cells during Agrobacterium infection might contribute to better understanding of basic biological processes as cell communication, intracellular transport and DNA repair and recombination as well as to expanding the host range of Agrobacterium as a genetic engineering tool.     

Carbon isotope ratios and the variation in the diurnal pattern of malate accumulation in aerial roots of CAM species of <Emphasis Type="Italic">Phalaenopsis</Emphasis> (Orchidaceae)

We investigated the carbon isotope ratios and the diurnal pattern of malate accumulation in leaves and aerial roots of eight species of Phalaenopsis grown in greenhouses. The leaves of all the species showed carbon isotope ratios and the diurnal patterns of malate content typical of CAM plants. However, the aerial roots exhibited a large variation in the diurnal pattern of malate content among species and even among plants within the same species, although carbon isotope ratios were always CAM-like values. Some aerial roots showed the typical diurnal pattern of CAM, but others maintained high or low malate contents during a day without fluctuation. In order to characterize more strictly the nature of the malate variation in the aerial roots, we further investigated a possible variation of the diurnal pattern of malate among different aerial roots within an individual for Phalaenopsis amabilis and P. cornu-cervi. The diurnal pattern of malate content was varied even among different aerial roots within the same plant. Thus the photosynthetic carbon metabolism in aerial roots of orchids is fairly complex.     

Effect of wounding on chalcone synthase and pathogenesis related <Emphasis Type="Italic">PR-10</Emphasis> gene expression and content of phenolic compounds in bilberry leaves

The influence of artificial wounding on biosynthesis of flavonoids and hydroxycinnamic acids was studied in bilberry leaves using two separate wounding experiments. In the first experiment bilberry leaves were wounded by cutting. The expression of the first gene from flavonoid pathway, chalcone synthase (CHS) and a wound induced pathogenesis related PR-10 gene was analysed from samples collected immediately and after 3, 6, 24 h and 4 d from the wounding treatment. In the second experiment annual shoots were removed. Proanthocyanidins, flavonol glycosides and hydroxycinnamic acids were quantified in leaf samples after 0–5 d (experiment 1) and 5 weeks (experiment 2) from the treatment. In the first experiment, no change was observed in the expression of CHS whereas increase in expression of PR-10 gene was detected after 6 h of wounding treatment. In both experiments, the contents of flavonol glycosides and hydroxycinnamic acids were not influenced by the wounding treatment and the contents of proanthocyanidins were decreased.     

Exogenous salicylic acid alleviates NaCl toxicity and increases antioxidative enzyme activity in <Emphasis Type="Italic">Lycopersicon esculentum</Emphasis>

Effects of exogenous salicylic acid (SA) on plant growth, contents of Na, K, Ca and Mg, activities of superoxide dismutase (SOD), guaiacol peroxidase (GPX), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR) and catalase (CAT), and contents of ascorbate and glutathione were investigated in tomato (Lycopersicon esculentum L.) plants treated with 100 mM NaCl. NaCl treatment significantly increased H₂O₂ content and lipid peroxidation indicated by accumulation of thiobarbituric acid reactive substances (TBARS). A foliar spray of 1 mM SA significantly decreased lipid peroxidation caused by NaCl and improved the plant growth. This alleviation of NaCl toxicity by SA was related to decreases in Na contents, increases in K and Mg contents in shoots and roots, and increases in the activities of SOD, CAT, GPX and DHAR and the contents of ascorbate and glutathione.     

Aerenchyma formation in maize roots

Maize (Zea mays L.) is generally considered to be a plant with aerenchyma formation inducible by environmental conditions. In our study, young maize plants, cultivated in various ways in order to minimise the stressing effect of hypoxia, flooding, mechanical impedance or nutrient starvation, were examined for the presence of aerenchyma in their primary roots. The area of aerenchyma in the root cortex was correlated with the root length. Although 12 different maize accessions were used, no plants without aerenchyma were acquired until an ethylene synthesis inhibitor was employed. Using an ACC-synthase inhibitor, it was confirmed that the aerenchyma formation is ethylene-regulated and dependent on irradiance. The presence of TUNEL-positive nuclei and ultrastructural changes in cortical cells suggest a connection between ethylene-dependent aerenchyma formation and programmed cell death. Position of cells with TUNEL-positive nuclei in relation to aerenchyma-channels was described.     

Lead uptake,toxicity and accumulation in <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> plants

The effects of lead were investigated in bean plants (Phaseolus vulgaris L. cv. Zlota Saxa) grown hydroponically in nutrient solution and exposed to Pb(NO₃)₂ (0.1, 0.5, 1 mM) with or without equimolar concentrations of chelator ethylenediaminetetraacetic acid (EDTA). The roots treated only with Pb(NO₃)₂ accumulated up to 25 g(Pb) kg⁻¹(d.m.), during 4-d exposure. However, in bean plants exposed to 0.5 mM Pb + 0.5 mM EDTA or 1 mM Pb + 1 mM EDTA 2.5 times less Pb was determined. In bean plants treated only with Pb, less than 6 % of total lead accumulated was transported to the aboveground parts, while in the case of plants grown with Pb + EDTA, around 50 % of total Pb was transported to the shoots.     

Genetic diversity assessment in Portugal accessions of <Emphasis Type="Italic">Olea europaea</Emphasis> by RAPD markers

Eighty seven olive (Olea europaea ssp. sativa L.) cultivar accessions from Portugal were characterized by means of randomly amplified polymorphic DNA (RAPD) markers. Of the 11 arbitrary 10-mer primers tested a total of 92 polymorphic bands were obtained, representing 87.6 % of the total amplification products. Twenty nine different genotypes were clearly discriminated. Differences were not found among the amplification profiles from different individuals of the same cultivar. All the genotypes could be identified by the combination of three primers: OPR-1, OPK-14 and OPA-1, seven genotype-specific markers being detected. Genetic relationships were estimated by the unweighted pair-group method with arithmetic averaging (UPGMA). The genetic analysis of the results showed a gradual distance between the various cultivars, making it difficult to identify well-differentiated phylogenetic groups, although two clusters were distinguishable with 35 % similarity, in addition to three independent branches with lower similarity: Galega, Tentilheira and Redondal. The dendrogram reflect some relationships for most of the cultivars according to the use of the fruit and ecological adaptation.