Effect of photosynthesis on dark mitochondrial respiration in green cells

Green plant cells can generate ATP in both chloroplasts and mitochondria. Hence the effect of photosynthesis on dark mitochondrial respiration can be considered at a variety of levels. Turnover of ceitric acid cycle dehydrogenases, which is essential for supply of carbon skeletons for amino acid synthesis, seems to be largely unaffected during photosynthesis. The source of carbon for the anaplerotic function of the citric acid cycle in light is however, not known with certainty. NADH generated in these reactions is probably not oxidised via the mitochondrial electron transfer chain coupled to ATP synthesis. However, it may be oxidised by the alternative cyanide-insensitive pathway, exported to the cytosol via the oxaloacetate-malate dicarboxylate shuttle or directly utilised for cytosolic nitrate reduction. Oxidation of succinate via cytochrome oxidase may also be similarly inhibited in light. Whether increase in the cytosolic ATP/ADP ratio in light is responsible for the inhibition of mitochondrial electron transfer to O₂ is not clearly established, because the ATP/ADP ratio is reported to be already quite high in the dark. Effective collaboration between photophosphorylation and oxidative phosphorylation in order to maintain the cytosolic energy charge at a present high level is discussed.     

高压静电场(HVEF)对柑桔青、绿霉病菌的抑制效果

用高压静电场(HVEF)对柑桔青霉菌 (P.italicum )和绿霉菌 (P.digitatum)分别进行了不同时间和方法的处理 ,以比较这些处理对病菌孢子在不同温度下萌发力和致病力的抑制效果。结果表明 ,HVEF处理后病菌孢子的萌发率、芽管长度和产孢量都显著地低于对照。而且处理时间越长 ,效果越明显。处理的孢子接种在柑桔果实上的感病率也显著低于对照 ,并能有效地抑制感病果实病斑的扩展。     

Can the fluorescence of green fluorescent protein chromophore be related directly to the nativity of protein structure?

In studies of green fluorescence protein (GFP) or other proteins with the use of GFP as a marker, the fluorescence of GFP is for the most part related directly to the nativity of its structure. Naturally, such a relation does exist since the chromophore of this protein is formed autocatalytically only just after GFP acquires its native structure. However, the fluorescence method may not yield reliable information on protein structure when studying renaturation and denaturation of this protein (with the formed chromophore). Using proteolysis, denaturant gradient gel electrophoresis and circular dichroism, we demonstrate herein that at major disturbances of the native structure of protein GFP-cycle3 the intensity of fluorescence of its chromophore can change insignificantly. In other words, the chromophore fluorescence does not reliably mirror alterations in protein structure. Since the main conclusions of this study are especially qualitative, it can be suggested that during renaturation/denaturation of wild-type GFP and its “multicolored” mutants their fluorescence is also not always associated with the changes in the structure of these proteins.     

Development of a new protein labeling system to map subunits and domains of macromolecular complexes for electron microscopy

Several gene fusion technologies have been successfully applied to label particular subunits or domains within macromolecular complexes to enable positional mapping of electron microscopy (EM) density maps, but exogenous fusion of a protein domain into the target polypeptide can cause unwanted structural and functional outcomes. Fab fragments from antibodies can be used as labeling reagents during EM visualization without gene manipulation of the target protein, but this method requires a panel of high-affinity antibodies that recognize a wide variety of epitopes. Linear peptide tags and their anti-tag antibodies can be used but they have a limited mapping ability as their placement is usually limited to the terminal regions of a protein. The PA dodecapeptide epitope tag (GVAMPGAEDDVV), forms a tight β-turn in the antigen binding pocket of its antibody (NZ-1). This capability allows for insertion of the PA tag into various surface-exposed loops within a multi-domain cell adhesion receptor, α_IIbβ₃ integrin. We confirmed that the purified PA-tagged integrin ectodomain fragments can form a stable complex with NZ-1 Fab. Negative stain EM of the various integrin-NZ-1 complexes revealed that a majority of the particles exhibited a clear density corresponding to the NZ-1 Fab; and the positions of the bound Fab were in good agreement with the predicted location of the inserted PA tag. The high-affinity and insertion-compatibility of the PA tag system allowed us to develop a new EM labeling methodology applicable to proteins for which good antibodies are not available.     

<Emphasis Type="Italic">Chlorobaculum tepidum</Emphasis> regulates chlorosome structure and function in response to temperature and electron donor availability

Green sulfur bacteria (GSB) rely on the chlorosome, a light-harvesting apparatus comprised almost entirely of self-organizing arrays of bacteriochlorophyll (BChl) molecules, to harvest light energy and pass it to the reaction center. In Chlorobaculum tepidum, over 97% of the total BChl is made up of a mixture of four BChl c homologs in the chlorosome that differ in the number and identity of alkyl side chains attached to the chlorin ring. C. tepidum has been reported to vary the distribution of BChl c homologs with growth light intensity, with the highest degree of BChl c alkylation observed under low-light conditions. Here, we provide evidence that this functional response at the level of the chlorosome can be induced not only by light intensity, but also by temperature and a mutation that prevents phototrophic thiosulfate oxidation. Furthermore, we show that in conjunction with these functional adjustments, the fraction of cellular volume occupied by chlorosomes was altered in response to environmental conditions that perturb the balance between energy absorbed by the light-harvesting apparatus and energy utilized by downstream metabolic reactions.     

Vertical distribution of some piercing sucking insects on some roselle varieties in Egypt and the role of amino acids concentration in infestation

Abstract

The cotton aphid, Aphis gossypii Glover was found to be the only aphid species infesting the tested roselle varieties (Sudani, Masri and White), Hibiscus sabdariffa L. which were cultivated in El-Kanater El-khayria (about 30 km north Cairo) as ex situ old land. The vertical distribution of the whitefly, Bemisia tabaci (Genn.); the leafhopper, Empoasca spp. and the pink hibiscus mealy bug, Maconellicoccus hirsutus Green, as insect pests attacking this crop had been studied. Moreover, certain morphological characters and amino acids concentrations in the three varieties of roselle, were obtained. The obtained results indicated that the experimental insect pests severely attacked leaves of the stem nodes from the 8th to the 11th (counting from the top of the plant), whereas whitefly were more abundant on these nodes. All tested insect species were less abundant on the 4th stem node. This may be due to the extensive existence of gland hairs, that excrete some compound especially phenols that might prevent the insects from reproducing on the leaves. The opposite was, however, true for M. hirsutus as it did not attack Sudani and Masri varieties but the infestation occurred at the highest level on the White variety.     

Evaluation of the bitterness of green tea catechins by a cell-based assay with the human bitter taste receptor hTAS2R39

Catechins have a broad range of physiological functions and act as the main taste ingredient of green tea. Although catechins show a strong bitterness, the bitter taste receptor for catechins has not been fully understood. The objective of this study was to identify the receptor for the major green tea catechins such as (−)-epicatechin (EC), (−)-epicatechin gallate (ECg), (−)-epigallocatechin (EGC), and (−)-epigallocatechin gallate (EGCg). By the cell-based assay using cultured cells expressing human bitter taste receptor, a clear response of hTAS2R39-expressing cells was observed to 300 μM of either ECg or EGCg, which elicit a strong bitterness in humans. The response of hTAS2R39-expressing cells to ECg was the strongest among the tested catechins, followed by EGCg. Because the cellular response to EC and EGC is much weaker than those of ECg and EGCg, galloyl groups was strongly supposed to be involved in the bitter intensity. This finding is similar to the observations of taste intensity obtained from a human sensory study. Our results suggest the participation of hTAS2R39 in the detection of catechins in humans, indicating the possibility that bitterness of tea catechins can be evaluated by using cells expressing hTAS2R39.     

Kinetic parameters of nitrogen mineralization rates of leguminous crops incorporated into soil

Summary Fresh leguminous plant residues were incorporated into soil columns and incubated at 23°C for up to 20 weeks. The N released from specific fractions (foliage, stems, and roots) of each residue were monitored at specific time intervals. Relationships between organic carbon, total nitrogen, CN ratio, lipids, and lignin content of the plant materials and the cumulative amount of N mineralized in soil were investigated. Statistical analyses indicated that the rates of N mineralized were not significantly correlated with the organic C nor lipid content of the residues. However, the cumulative amount of N released was significantly correlated with the total N content of the plant material (r=0.93^***). The percentage of organic N of the legumes mineralized in soil ranged from 15.9 to 76.0%. The relationship between the percentage of N released and the CN ratio of the plant material showed an inverse cuvilinear response (r= 0.88^***). It was also evident that the composition of lignin in the residue influenced N mine-ralization rates of the leguminous organs incorporated into soil.There was a curvilinear relationship between the cumulative amount of N released from the residues and time of incubation. Nitrogen mineralization rates were described by first-order kinetics to estimate the N mineralization potential (N₀), mineralization rate constant (k), and the time of incubation required to mineralize one-half of N₀ (t_1/2). The kinetic parameters were calculated by both the linear least squares (LLS) and nonlinear least squares (NLLS) transformations. The N₀ values among the crop residues varied from –35 to 510 g Ng^–1 soil. Statistical analyses revealed that the N₀ values obtained by both LLS and NLLS methods were significantly correlated (r=0.93^***). The mineralization rate constants (k) of the residues ranged from 0.045 to 0.325 week^–1. The time of incubation required to mineralize one-half the nitrogen mineralization potential (t_1/2) of the legumes incorporated into soil ranged from 2.1 to 15.4 weeks.     

Prediction and interpretation of the antioxidant capacity of green tea from dissimilar chromatographic fingerprints

Previously, multivariate calibration techniques have been successfully applied to model and predict the antioxidant activity of green tea from its chromatographic fingerprint. Since the selectivity differences between dissimilar chromatographic systems have already been valuably used in several applications, in this paper it is studied whether combining the complementary information contained in two dissimilar fingerprints can improve the predictive capacity of the multivariate calibration model. The simplest way of combining the data is concatenating both fingerprints for each sample. The resulting matrix can then be subjected to Orthogonal Projections to Latent Structures (O-PLS). Unfortunately, this approach resulted in a more complex model with a prediction error of about the average of the errors obtained with the individual fingerprints. Secondly, only the peaks with high loading and low orthogonal loading from both chromatograms were included in the O-PLS model. This resulted in a reduced complexity, but not in better predictions, probably due to a lack of complementarity of the information concerning the antioxidant capacity. Finally, the concatenated fingerprints were subjected to stepwise multiple linear regression (MLR) in order to build a model based on the variables most correlated with the antioxidant capacity. The obtained prediction error was lower than those of both previous approaches, but still higher than the error of the model based on a single analysis. This is probably again caused by a lack of complementarity in the variables. Nevertheless, it was advantageous to develop fingerprints on dissimilar system, because it enables to choose the most suited chromatographic profile to build a multivariate calibration model for the considered purpose. In contrast to what was expected, the study showed that the most simple (so the worst separated) fingerprints resulted in the best predictions. On the other hand, a more complex fingerprint in which more compounds are separated is still important to improve the interpretability of the model.     

Limnology of the Green Lakes Valley: phytoplankton ecology and dissolved organic matter biogeochemistry at a long-term ecological research site

ABSTRACT

Background: Surface waters are the lowest points in the landscape, and therefore serve as excellent integrators and indicators of changes taking place in the surrounding terrestrial and atmospheric environment.

Aims: Here we synthesise the findings of limnological studies conducted during the past 15 years in streams and lakes in the Green Lakes Valley, which is part of the Niwot Ridge Long Term Ecological Research (LTER) Site.

Methods: The importance of these studies is discussed in the context of aquatic ecosystems as indicators, integrators, and regulators of environmental change. Specifically, investigations into climatic, hydrologic, and nutrient controls on present-day phytoplankton, and historical diatom, community composition in the alpine lake, Green Lake 4, are reviewed. In addition, studies of spatial and temporal patterns in dissolved organic matter (DOM) biogeochemistry and reactive transport modelling that have taken place in the Green Lakes Valley are highlighted.

Results and conclusions: The findings of these studies identify specific shifts in algal community composition and DOM biogeochemistry that are indicative of changing environmental conditions and provide a framework for detecting future environmental change in the Green Lakes Valley and in other alpine watersheds. Moreover, the studies summarised here demonstrate the importance of long-term monitoring programmes such as the LTER programme.