Photosystem 2-activity and thylakoid membrane polypeptides of <Emphasis Type="Italic">in vitro</Emphasis> cultured chrysanthemum as affected by NaCl

Long-term (30 d) effects of 100, 200, 300, and 400 mM NaCl on photosystem 2 (PS 2)-mediated electron transport activity and content of D1 protein in the thylakoid membranes of chrysanthemum (Dendranthema grandiflorum) cultured in vitro at low irradiance 20 μmol(photon) m⁻² s⁻¹ were investigated. 100 mM NaCl increased contents of chlorophylls (Chl) a and b, carotenoids (Car; xanthophylls + carotenes), and the ratio of Chl a/b, and Car/Chl a+b. However, further increase in NaCl concentration led to the significant reduction in the contents of Chl a, and Chl b, and increase in the ratio of Chl a/b and Car/Chl a+b. NaCl treatment decreased the PS 2-mediated electron transport activity and contents of various thylakoid membrane polypeptides including D1 protein.     

Stress-induced degradation of D1 protein and its photoprotection by DCPIP in isolated thylakoid membranes of barley leaf

Effects of various stress treatments such as NaCl, hydrogen peroxide, hydroxyl free radical, and high irradiance (HI, 1 000 μmol m⁻² s⁻¹) on the photosystem (PS) 2 mediated electron transport rate and the degradation of D1 protein in the thylakoid membranes of barley were studied. The applied stresses caused significant reduction in the PS 2-mediated electron transport and a degradation of D1 protein that was highest during the HI-treatment. Presence of 2,6-dichlorophenol indophenol (DCPIP), which is an artificial electron acceptor from water, significantly minimizes the HI-induced deleterious effect on the PS 2-mediated electron transport rate, disarrangement of PS machinery, and degradation of the D1 protein. HI in the absence of an acceptor resulted in production of reactive oxygen species due to electron transfer to oxygen.     

Glycine betaine improves thylakoid membrane function of tobacco leaves under low-temperature stress

Glycine betaine (GB) is an effective compatible solute that improves the tolerance in plants to various stresses. We investigated the effects of 2 mM GB applied to the roots of a tobacco (Nicotiana tabacum L.) cultivar on enhancing photosynthesis under low-temperature (LT) stress (5/5 °C, 12/12 h, 300 μmol m⁻² s⁻¹) and in the subsequent recovery (25/18 °C) from the stress. The net photosynthetic rate, intrinsic efficiency measured as the ratio of variable to maximum fluorescence, and actual efficiency of the photochemistry of photosystem 2 as well as the ATPase activity in the thylakoid membrane decreased, and a distinct K step in the fluorescence transient O-J-I-P appeared under cold stress. Exogenous GB alleviated the decrease in all these parameters. The LT-stress induced the accumulation of 33–66 kDa polypeptides and decreased the proportion of unsaturated fatty acids in the thylakoid membrane. In plants subjected to LT-stress, GB protected these polypeptides from damage and enhanced the proportion of unsaturated fatty acids. An increase in non-radiative energy dissipation (NPQ) may be involved in the improvement of the function of the thylakoid membrane by GB since exogenous GB protected violaxanthin de-epoxidase and enhanced NPQ.     

Polyamine metabolism in hants : Arginine and omithine decarboxylase activity in ripeningTrlticum durum seeds

The pattern of the activity of arginine decarboxylase (ADC) and omithine decarboxylase (ODC) involved in polyamine synthesis in ripening wheat seeds was examined. The aim was to study the polyamines and the activity of the two enzymes in correlation with the growth processes occurring in the developing wheat seeds. The results obtained showed a very different pattern of polyamine content in the two organs of caryopsis, and that the two enzymes in the embryos have a higher activity than in the endosperms. Moreover, while in the embryos the ADC exhibits higher activity than the ODC, in the endosperms the activity of ODC is about similar to that of ADC. This pattern is discussed in relation to the different histological characteristics of embryo and endosperm tissues during seed development.     

Effect of osmotic stress on transpiration and absorption rates in triticale and its parental species

In triticale and its parental species, the application of a root osmotic stress induced either a transient increase or an immediate decrease in transpiration rate. The response of wheat (Trticum dicoccum farrum) proved to depend on relative humidity of air. In rye (Secale cerealecv. Petkus) and triticale (T. 300) the effect of NaCl stress was less expressive, than the effect of PEG.     

Response of senescing rice leaves to flooding stress

Flooding stress (FS) induced changes in pigment and protein contents and in photochemical efficiency of thylakoid membranes of chloroplasts were investigated during senescence of primary leaves of rice seedlings. Leaf senescence was accompanied by loss in 2,6-dichlorophenolindophenol (DCPIP) photoreduction, rate of oxygen evolution, quantum yield of photosystem 2 with an increase in MDA accumulation, and non-photochemical quenching (NPQ) of chlorophyll fluorescence. These changes were further aggravated when the leaves during this period experienced FS. The increase in NPQ value under stress may indicate photosynthetic adaptation to FS.     

Effects of evening and nighttime leaf wetting on stomatal behavior of Cryptomeria japonica growing in dry soil

To examine the hypothesis that stomatal behavior of plants in dry soil is influenced by a slow recovery from daytime water deficit, we studied the effect of repeated wetting of leaves during evening and night in Cryptomeria japonica seedlings grown in dry soil. After 7 and 10 days of leaf wetting treatment the midday leaf water potential decreased and the transpiration rate increased, respectively. Therefore, we suggest that rapid recovery from daytime water deficit could weaken the water conserving stomatal behavior that adapts to drought conditions in the roots.     

Diurnal changes in chlorophyll fluorescence and light utilization in Colocasia esculenta leaves grown in marshy waterlogged area

Diurnal cycle of chlorophyll fluorescence parameters was done in Colocasia esculenta L. (swamp taro) grown in marshy land under sun or under shade. The sun leaves maintained higher electron transport rate (ETR) and steady state to initial fluorescence ratio (F_s/F₀) than shade leaves. In spite of lower ETR, higher photochemical quenching (PQ), and effective quantum yield of photosystem 2 (Φ_PS2) was evident in shade plants compared to plants exposed to higher irradiance. ETR increased linearly with increase in irradiance more under low irradiance (r ² = 0.84) compared to higher irradiance (r ² = 0.62). The maximum quantum yield of PS 2 (F_v/F_m) did not differ much in sun and shade leaves with the exception of midday when excess of light energy absorbed by plants under sun was thermally dissipated. Hence swamp taro plants adopted different strategies to utilize radiation under different irradiances. At higher irradiance, there was faster decline in proportion of open PS 2 centers (PQ) and excess light energy was dissipated through non-photochemical quenching (NPQ). Under shade, absorbed energy was effectively utilized resulting in higher Φ_PS2.     

Cadmium effects on the organization of microtubular cytoskeleton in interphase and mitotic cells of Allium sativum

We have investigated the effects of cadmium on the microtubular (MT) cytoskeleton in the root tip cells of Allium sativum L. using indirect immunofluorescence microscopy. Cd affected the mechanisms controlling the organization of MT cytoskeleton, as well as tubulin assembly/disassembly processes. Cd induced the formation of abnormal MT arrays, consisting of discontinuous wavy MTs or short MT fragments at the cell periphery. Cadmium caused irregular nuclear disorder in cells where the MT organization and function was disturbed. Furthermore, with increased Cd concentration and duration of treatment the MTs depolymerized more severely, the frequency of abnormal cell increased and the mitotic index decreased progressively. The above findings showed that MT cytoskeleton is one of target sites of Cd toxicity in root tip cells.     

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.     

Photosynthetic behaviour of Arabidopsis plants with a Cap Binding Protein 20 mutation under water stress

Under non-stressed conditions the net photosynthetic rate (P _N) of the mutant plants cbp20 of Arabidopsis was similar to that of the wild type (WT). In response to water deprivation, however, P _N started to decrease later in the mutants and remained substantially higher. Thermoluminescence measurements showed that the lipid peroxidation induced by severe water stress was also less pronounced in the mutant than in the WT. Both soil gravimetric and plant water potential data showed that cbp20 mutants lose water more slowly than the WT plants. The drought-induced decline in F_v/F_m, the quantum efficiency of photosystem 2, and photochemical quenching parameters also started later in the cbp20 mutants than in the WT plants. Thus the restricted gas exchange in the cbp20 mutants does not impair the photosynthetic performance of the plant; however, under drought improved water retention provides significant protection for the photosynthetic apparatus.     

Ozone sensitivity and ethylenediurea protection in ash trees assessed by JIP chlorophyll <Emphasis Type="Italic">a</Emphasis> fluorescence transient analysis

The effect of ethylenediurea (EDU) was tested using the chlorophyll (Chl) a fluorescence transient analysis, performed with JIP-test, to assess ambient ozone (O₃) effects on photosynthesis of adult trees under natural conditions. Twelve adult European ash (Fraxinus excelsior L.) trees, known to be sensitive or tolerant to O₃, determined by presence symptomatic (S) or absence asymptomatic (AS) trees of foliar symptoms in previous years, were treated either with distilled water containing 450 g m⁻³ EDU or with distilled water. Once a month across the growing season [the accumulated exposure over a threshold of 40 nmol(O₃) mol⁻¹ was 32.49 μmol mol⁻¹ h⁻¹], Chl a fluorescence transients were measured in vivo on dark-adapted leaves of 1-year-old labeled shoots, from the lower crown part. Twenty-five parameters were calculated. The maximum quantum yield of primary photochemistry (ϕ_Po or F_v/F_m) did not differentiate between S-and AS-trees, while increased Chl content and de-excitation rates suggested compensation of O₃ injury in S-trees. Seasonal reductions in absorbing fluxes and increase in heat and fluorescence dissipation processes was due to leaf ageing and drought, the latter suggesting water deficit influenced Chl a fluorescence stronger than ambient O₃ exposure. AS-trees showed elevated probability of connectivity among photosystem 2 units, a mechanism to stimulate energy dissipation and reduce photo-oxidative injury. EDU prevented the inactivation of reaction centers. This slight effect does not warrant EDU as a tool to assess O₃ effects on photosynthesis, while the JIP-test is suggested for a quantitative assessment in adult trees.     

Genetic analysis of fertility-restorer genes in rice

Wild-abortive (WA), Honglian (HL) and Baro-II (BT) are three important cytoplasmic male sterility (CMS) types in rice. It is essential to investigate genetic mode and allelism of fertility restorer (Rf) genes and the relationship between Rf and CMS. Fertility of the all test-cross F₁ plants shows that the restorer-maintainer relationship is similar for HL-CMS and BT-CMS, while that is variance for WA-CMS and HL-CMS (or BT-CMS), respectively. Genetic analysis of Rf genes indicates that HL-or BT-CMS are controlled by single dominant Rf gene and WA-CMS is controlled by one or two pairs of dominant Rf genes, which reflects the characters of the gametophytic and sporophytic restoration CMS type. It is concluded that there are at least three Rf loci in different accessions with Rf genes for each CMS type.     

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.     

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.     

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.     

Measuring solution viscosity and its effect on enzyme activity

In proteins, some processes require conformational changes involving structural domain diffusion. Among these processes are protein folding, unfolding and enzyme catalysis. During catalysis some enzymes undergo large conformational changes as they progress through the catalytic cycle. According to Kramers theory, solvent viscosity results in friction against proteins in solution, and this should result in decreased motion, inhibiting catalysis in motile enzymes. Solution viscosity was increased by adding increasing concentrations of glycerol, sucrose and trehalose, resulting in a decrease in the reaction rate of the H⁺-ATPase from the plasma membrane ofKluyveromyces lactis. A direct correlation was found between viscosity (η) and the inhibition of the maximum rate of catalysis (V _max). The protocol used to measure viscosity by means of a falling ball type viscometer is described, together with the determination of enzyme kinetics and the application of Kramers’ equation to evaluate the effect of viscosity on the rate of ATP hydrolysis by the H⁺-ATPase. Published: May 1, 2003