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Separate action spectra for the two photochemical systems of photosynthesis.

Action spectra have been obtained which are believed to represent the 2 photochemical systems in the marine green alga, Ulva lobata. Measurement of transient rates of O(2) uptake in the presence of DCMU furnished the data for the spectrum representing system I. The action spectrum for system II refers to O(2) evolution rates after the Ulva was pretreated in an atmosphere of high O(2) concentration. Tissue responses after high O(2) exposure suggests an inhibition of the photosynthetic functioning of system I.

Journal Article↗

Physiological effects of the presence and absence of gas vacuoles in the blue-green alga, Microcystis aeruginosa Kuetz. emend. Elenkin.

Physiological evidence was obtained for a light shielding role for gas vacuoles in Microcystis aeruginosa Kuetz. emend. Elenkin, by comparing photosynthetic oxygen evolution, growth behaviour and pigment composition of cells with intact or collapsed gas vacuoles. The oxygen evolution rates were strongly dependent on cell concentration, a maximum rate for cells with intact gas vacuoles occurring at about 1.4 X 10(9) cells/ml and for cells with collapsed gas vacuoles at about 2.5 X 10(9) cells/ml. By using light saturation curves for oxygen evolution, it was estimated that at low light intensities up to 30% of the photosynthetically useable light was shielded at a cell concentration of 6 X 10(8) cells/ml. Collapsing the gas vacuoles twice daily did not alter the initial growth rate of the cultures, but enabled them to reach a higher final cell density. Collapsing of gas vacuoles during growth for about four generations resulted in a lower level of all acetone soluble pigments with a greater relative reduction in carotenoids than in chlorophyll a. Collapse of the gas vacuoles does not alter the cell volume. Various optical interactions which could account for light shielding are discussed.

Light↗

Endogenous indole-3-acetic acid and ethylene evolution in tilted Metasequoia glyptostroboides stems in relation to compression-wood formation.

Eight-year-old Metasequoia glyptostroboides seedlings were tilted at a 45 degrees angle to induce compression-wood formation on the lower side of the stems. After 2 weeks of treatment, half of the seedlings were sampled and the remaining half were tilted to the opposite orientation to exchange the upper and lower sides and were kept for 2 more weeks until sampled. Cambium-emitted ethylene was analyzed by gas chromatography with flame-ionization detection. Endogenous indole-3-acetic acid (IAA) was measured by gas chromatography-mass spectrometry. Tracheid production and compression-wood formation were determined by light microscopy. Anatomical studies showed that tracheid production was promoted and compression-wood tracheids always developed on the gravitationally lower side of tilted stems in both the original tilting and the subsequent reverse-tilting periods. These were accompanied by an increase in IAA content in and an accelerated ethylene-evolution rate from the cambial region of the same side.

Biological Evolution↗

Metabolic regulation analysis of icd-gene knockout Escherichia coli based on 2D electrophoresis with MALDI-TOF mass spectrometry and enzyme activity measurements.

An integrated study of cell growth characteristics, enzyme activities and protein expression patterns was carried out to investigate how the central metabolism of Escherichia coli changes upon knockout of the isocitrate dehydrogenase (ICDH) gene (icd) in the tricarboxylic acid cycle. Deletion of the icd gene led to reduced specific growth rate and reduced specific glucose consumption rate. The reduced specific growth rate in the icd mutant was due mainly to the lower intracellular ATP/ADP ratio as well as to the lower NADPH/NADP(+) ratio compared with those in the parent strain. However, the specific carbon dioxide evolution rate was found to be higher in the icd mutant strain compared to the parent E. coli. This may be due to the higher activity of 6-phosphogluconate dehydrogenase, phosphoenol pyruvate carboxykinase and NADP(+)-dependent malic enzymes. The glyoxylate pathway was also utilized, as evidenced by the significant upregulation of isocitrate lyase and malate synthase activity in the icd mutant E. coli. The appearance of the glyoxylate pathway caused lower acetate production. Of 21 proteins showing altered expression levels, 17 were successfully identified with the aid of MALDI-TOF mass spectrometry. The results showed that the abolition of ICDH activity significantly affected the respiratory system and electron transport chain, as evidenced by the significant downregulation of proteins encoded by the genes nuoE, nuoH, cydA and cyoA in icd mutant E. coli compared to the parent.

Carbohydrate Metabolism↗

Modelling Yersinia enterocolitica inactivation in coculture experiments with Lactobacillus sakei as based on pH and lactic acid profiles.

In food processing and preservation technology, models describing microbial proliferation in food products are a helpful tool to predict the microbial food safety and shelf life. In general, the available models consider microorganisms in pure culture. Thus, microbial interactions are ignored, which may lead to a discrepancy between model predictions and the actual microbial evolution, particularly for fermented and minimally processed food products in which a background flora is often present. In this study, the lactic acid mediated negative microbial interaction between the lactic acid bacterium Lactobacillus sakei and the psychrotrophic food pathogen Yersinia enterocolitica was examined. A model describing the lactic acid induced inhibition (i.e., early induction of the stationary phase) of the pathogen [Vereecken, K.M., Devlieghere, F., Bockstaele, A., Debevere, J., Van Impe, J.F., 2003. A model for lactic acid induced inhibition of Yersinia enterocolitica in mono- and coculture with Lactobacillus sakei. Food Microbiology 20, 701-713.] was extended to describe the subsequent inactivation (i.e., decrease of the cell concentration to values below the detection limit). In the development of a suitable model structure to describe the inactivation process, critical points in the variation of the specific evolution rate mu [1/h] with the dynamic (time-varying) pH and undissociated lactic acid profiles were taken into account. Thus, biological knowledge, namely, both pH and undissociated lactic acid have an influence on the microbial evolution, was incorporated. The extended model was carefully validated on new data. As a result, the newly developed model is able to accurately predict the growth, inhibition and subsequent inactivation of Y. enterocolitica in coculture as based on the dynamic pH and lactic acid profiles of the medium.

Antibiosis↗

Physiological response to anaerobicity of glycerol-3-phosphate dehydrogenase mutants of Saccharomyces cerevisiae.

Mutants of Saccharomyces cerevisiae, in which one or both of the genes encoding the two isoforms of NAD-dependent glycerol-3-phosphate dehydrogenase had been deleted, were studied in aerobic batch cultures and in aerobic-anaerobic step change experiments. The respirofermentative growth rates under aerobic conditions with semisynthetic medium (20 g of glucose per liter) of two single mutants, gpd1 delta and gpd2 delta, and the parental strain (mu = 0.5 h-1) were almost identical, whereas the growth rate of a double mutant, gpd1 delta gpd2 delta, was approximately half that of the parental strain. Upon a step change from aerobic to anaerobic conditions in the exponential growth phase, the specific carbon dioxide evolution rates (CER) of the wild-type strain and the gpd1 delta strain were almost unchanged. The gpd2 delta mutant showed an immediate, large (> 50%) decrease in CER upon a change to anaerobic conditions. However, after about 45 min the CER increased again, although not to the same level as under aerobic conditions. The gpd1 delta gpd2 delta mutant showed a drastic fermentation rate decrease upon a transition to anaerobic conditions. However, the CER values increased to and even exceeded the aerobic levels after the addition of acetoin. High-pressure liquid chromatographic analyses demonstrated that the added acetoin served as an acceptor of reducing equivalents by being reduced to butanediol. The results clearly show the necessity of glycerol formation as a redox sink for S. cerevisiae under anaerobic conditions.

Acetoin↗

A rigorous method for multigenic families' functional annotation: the peptidyl arginine deiminase (PADs) proteins family example.

BACKGROUND: large scale and reliable proteins' functional annotation is a major challenge in modern biology. Phylogenetic analyses have been shown to be important for such tasks. However, up to now, phylogenetic annotation did not take into account expression data (i.e. ESTs, Microarrays, SAGE, ...). Therefore, integrating such data, like ESTs in phylogenetic annotation could be a major advance in post genomic analyses. We developed an approach enabling the combination of expression data and phylogenetic analysis. To illustrate our method, we used an example protein family, the peptidyl arginine deiminases (PADs), probably implied in Rheumatoid Arthritis. RESULTS: the analysis was performed as follows: we built a phylogeny of PAD proteins from the NCBI's NR protein database. We completed the phylogenetic reconstruction of PADs using an enlarged sequence database containing translations of ESTs contigs. We then extracted all corresponding expression data contained in EST database This analysis allowed us 1/To extend the spectrum of homologs-containing species and to improve the reconstruction of genes' evolutionary history. 2/To deduce an accurate gene expression pattern for each member of this protein family. 3/To show a correlation between paralogous sequences' evolution rate and pattern of tissular expression. CONCLUSION: coupling phylogenetic reconstruction and expression data is a promising way of analysis that could be applied to all multigenic families to investigate the relationship between molecular and transcriptional evolution and to improve functional annotation.

Animals↗

Comparative genetic diversity of the narG, nosZ, and 16S rRNA genes in fluorescent pseudomonads.

The diversity of the membrane-bound nitrate reductase (narG) and nitrous oxide reductase (nosZ) genes in fluorescent pseudomonads isolated from soil and rhizosphere environments was characterized together with that of the 16S rRNA gene by a PCR-restriction fragment length polymorphism assay. Fragments of 1,008 bp and 1,433 bp were amplified via PCR with primers specific for the narG and nosZ genes, respectively. The presence of the narG and nosZ genes in the bacterial strains was confirmed by hybridization of the genomic DNA and the PCR products with the corresponding probes. The ability of the strains to either reduce nitrate or totally dissimilate nitrogen was assessed. Overall, there was a good correspondence between the reductase activities and the presence of the corresponding genes. Distribution in the different ribotypes of strains harboring both the narG and nosZ genes and of strains missing both genes suggests that these two groups of strains had different evolutionary histories. Both dissimilatory genes showed high polymorphism, with similarity indexes (Jaccard) of between 0.04 and 0.8, whereas those of the 16S rRNA gene only varied from 0.77 to 0.99. No correlation between the similarity indexes of 16S rRNA and dissimilatory genes was seen, suggesting that the evolution rates of ribosomal and functional genes differ. Pairwise comparison of similarity indexes of the narG and nosZ genes led to the delineation of two types of strains. Within the first type, the similarity indexes of both genes varied in the same range, suggesting that these two genes have followed a similar evolution. Within the second type of strain, the range of variations was higher for the nosZ than for the narG gene, suggesting that these genes have had a different evolutionary rate.

DNA, Ribosomal↗

Generalized model and optimum performance of an irreversible quantum Brayton engine with spin systems.

The purpose of this paper is to establish a model of an irreversible quantum Brayton engine using many noninteracting spin systems as the working substance and consisting of two irreversible adiabatic and two isomagnetic field processes. The time evolution of the total magnetic moment M is determined by solving the generalized quantum master equation of an open system in the Heisenberg picture. The time of two irreversible adiabatic processes is considered based on finite-rate evolution. The relationship between the power output P and the efficiency eta for the irreversible quantum Brayton engine with spin systems is derived. The optimally operating region (or criteria) for the engine is determined. The influences of these important parameters on the performances (P and eta) of the engine are discussed. The results obtained herein will be useful for the further understanding and the selection of the optimal operating conditions for an irreversible quantum Brayton engine with spin systems.

Journal Article↗

Substitution of Ala-251 of the D1 reaction centre polypeptide with a charged residue results in impaired function of photosystem II.

Ala-251 in the membrane-parallel helix in the D-E loop of the D1 polypeptide close to the Q(B) pocket of photosystem II (PS II), was mutated to aspartate (D), lysine (K), leucine (L) or serine (S) in Synechocystis 6803. O2 evolution rates (H2O-->DCBQ; 2,6-dichloro-p-benzoquinone) of A251D, A251L and A251S were lower, being 38, 16, 62 and 70%, respectively, of that of the control, and there was an even more drastic impairment of O2 evolution when measured from H2O to DMBQ (2,5-dimethyl-p-benzoquinone), demonstrating modifications in the Q(B) pocket. However, in all other mutants but A251K, the Q(B) function could sustain O2 evolution at a level high enough to support photosynthetic growth. The mutant A251S, carrying a substitution of alanine for a chemically quite similar residue serine, was less severely affected. Substitution by a positively charged residue drastically delayed chlorophyll a fluorescence relaxation in the non-photosynthetic strain A251K, implying strong impairment of Q(A)-to-Q(B) electron transfer. Delay of fluorescence relaxation was clear in A251D as well, carrying a substitution of alanine for a negatively charged residue. The effects of the substitutions of A251 demonstrate the importance of this residue of the D1 polypeptide in the conformation of the acceptor side of PS II and, accordingly, the effect on the acceptor-side function of PS II was very clear. Nevertheless, the tolerance of PS II activity to high-light-induced photoinhibition in vivo and the subsequent D1 degradation were not much impaired in any of the photosynthetic mutant strains as compared to the control.

Alanine↗

Evolutionary systems biology: links between gene evolution and function.

The recent accumulation of genome-wide data on various facets of gene expression, function and evolution stimulated the emergence of a new field, evolutionary systems biology. Many significant correlations were detected between variables that characterize the functioning of a gene, such as expression level, knockout effect, connectivity of genetic and protein-protein interaction networks, and variables that describe gene evolution, such as sequence evolution rate and propensity for gene loss. The first attempts on multidimensional analysis of genomic data yielded composite variables that describe the 'status' of a gene in the genomic community. However, it remains uncertain whether different functional variables affect gene evolution synergistically or there is a single, dominant factor. The number of translation events, linked to selection for translational robustness, was proposed as a candidate for such a major determinant of protein evolution. These developments show that, although the methodological basis of evolutionary systems biology is not yet fully solidified, this area of research is already starting to yield fundamental biological insights.

Evolution, Molecular↗

Photorespiratory rates in wheat and maize as determined by o-labeling.

A method was devised to quantify short-term photorespiratory rates in terrestrial plants using (18)O-intermediates of the glycolate pathway, specifically glycolate, glycine, and serine. The pathway intermediates were isolated and analyzed on a GC/MS to determine molecular percent (18)O-enrichment. Rates of glycolate synthesis were determined from (18)O-labeling kinetics of the intermediates, derived rate equations, and nonlinear regression techniques. Glycolate synthesis in wheat (Triticum aestivum L.), a C(3) plant, and maize (Zea mays L.), a C(4) plant, was stimulated by high O(2) concentrations and inhibited by high CO(2) concentrations. The synthesis rates were 7.3, 2.1, and 0.7 micromoles per square decimeter per minute under a 21% O(2) and 0.035% CO(2) atmosphere for leaf tissue of wheat, maize seedlings, and 3-month-old maize, respectively. Photorespiratory CO(2) evolution rates were estimated to be 27, 6, and 2%, respectively, of net photosynthesis for the three groups of plants under the above atmosphere. The results from maize tissue support the hypothesis that C(4) plants photorespire, albeit at a reduced rate in comparison to C(3) plants, and that the CO(2)/O(2) ratio in the bundle sheath of maize is higher in mature tissue than in seedling tissue. The pool size of the three photorespiratory intermediates remained constant and were unaffected by changes in either CO(2) or O(2) concentrations throughout the 10-minute labeling period. This suggests that photorespiratory metabolism is regulated by other mechanism besides phosphoglycolate synthesis by ribulose-1,5-bisphosphate carboxylase/oxygenase, at least under short-term conditions. Other mechanisms could be alternate modes of synthesis of the intermediates, regulation of some of the enzymes of the photorespiratory pathway, or regulation of carbon flow between organelles involved in photorespiration. The glycolate pool became nearly 100% (18)O-labeled under an atmosphere of 40% O(2). This pool failed to become 100% (18)O-enriched under lower O(2) concentrations.

Journal Article↗

Speciation by host switch and adaptive radiation in a fish parasite genus Gyrodactylus (Monogenea, Gyrodactylidae).

Four hundred Gyrodactylus species have been formally described, but the estimated number of species in this fish ectoparasite genus of Monogenean Platyhelminthes is more than 20,000. The unusually high species richness has lead to the hypotheses of speciation and adaptive radiation via host switching. These hypotheses were tested by reconstructing a molecular phylogeny for the subgenus G. (Limnonephrotus) which is a group of freshwater parasites, including five species infecting wild and farmed salmonids. The highly variable ITS1 and ITS2 segments and the conservative 5.8S ribosomal gene were sequenced in 22 species plus two species representing the subgenus G. (Paranephrotus) as an outgroup. The phylogeny was compared with host systematics: the species were collected from six fish families (Cyprinidae, Salmonidae, Percidae, Esocidae, Gasterosteidae, and Gobitidae). The phylogenetic analysis demonstrated that G. (Limnonephrotus) is a monophyletic group that was originally hosted by cyprinids. The speciation has occurred in two episodes, the older one manifested in genetic distances 25-33% (4-6 Myr BP). The latter speciation burst occurred in one clade only, perhaps one million years ago. This clade has been morphologically identified as a wageneri species group. It is a monophyletic group of 18 species [studied here] and contains all five salmonid parasites, but also parasites, on cyprinids, percids, esocids, and gasterosteids. In G. (Limnonephrotus), eight host switches crossing the host family barrier were observed, and at least three of them were followed by repetitive speciation. Seven host-switch events were statistically confirmed by bootstrapping. The suggested model of speciation by host switch was accepted, and interestingly the adaptive radiation seems to be a consequence of host switch to a new family (key innovation model). The molecular and ecological evolution rate of Gyrodactylus parasites is manyfold in comparison to host species, and the phylogenies are largely independent and disconnected.

Animals↗

Comparative mapping of cosmids and gene clones from a 1.6 Mb chromosomal region of Drosophila melanogaster in three species of the distantly related subgenus Drosophila.

The successful hybridization of cosmid clones from Drosophila melanogaster (Sophophora subgenus) to the salivary gland chromosomes of other species as distantly related as those in the Drosophila subgenus attests their great potential for unravelling genome evolution. We have carried out, using 28 cosmids and 13 gene clones, a study of the organization of the D. melanogaster 95A-96A chromosomal region in three Drosophila subgenus species: D. repleta, D. buzzattii and D. virilis. These clones were first used to built an accurate map of this 1.6 Mb region of D. melanogaster chromosome 3R (Muller's element E). Then, they were hybridized and mapped to the homologous chromosome 2 of the other three distantly related species. The studied region is disseminated over 13 different sites of chromosome 2 in the Drosophila subgenus species, which implies a minimum of 12 inversion breakpoints fixed between the two subgenera. Extrapolation to the entire chromosome gives 90 fixed inversions. The D. melanogaster Pp1-96A-Acr96Aa segment conserved in D. repleta and D. buzzatii is longer than previously thought and is also conserved in D. virilis. In addition, three other D. melanogaster segments conserved in the three Drosophila subgenus species were found. Finally, our data indicate significant statistical differences in the evolution rate of Muller's element E among lineages, a result that agrees well with the previous cytogenetic data.

Animals↗

Physiological effects of nitrogen starvation in an anaerobic batch culture of Saccharomyces cerevisiae.

The effects of nitrogen starvation on the anaerobic physiology of Saccharomyces cerevisiae were studied in cells cultivated in a bioreactor. The composition of the mineral medium was designed such that the nitrogen source became depleted while there was still ample glucose left in the medium. The culture was characterized by acoustic gas analysis, flow injection analysis and HPLC analysis of extracellular substrates and metabolites. During the cultivation, the macromolecular composition of the cells was analysed with respect to the cellular content of RNA, protein, trehalose and glycogen. During exponential growth under anaerobic conditions, the maximum specific growth rate conditions. Depletion of ammonium in the medium led to an abrupt decrease (mumax) of S. cerevisiae CBS 8066 (0.46 h-1) was identical to the mumax determined under aerobic in the flux through glycolysis. Subsequently, a continuous decrease in the carbon dioxide evolution rate, caused by catabolite inactivation of the hexose-transport system, was observed. The apparent half-life of the transport system under nitrogen starvation was 13 h. During the exponential growth phase, the cellular content of RNA and protein was 15% (w/w) and 60% (w/w), respectively. At the end of the cultivation where the cells had been starved of nitrogen for 18 h, the cellular content of RNA and protein had decreased to 4% (w/w) and 22% (w/w), respectively. The intracellular carbohydrate content increased dramatically as trehalose and glycogen accumulated to final concentrations of 7% (w/w) and 25% (w/w), respectively. Glycerol formation during nitrogen starvation was higher than that accounted for by the formation of organic acids, suggesting a protein turnover of approximately 6% h-1. The growth energetics of S. cerevisiae CBS 8066 also changed as a result of nitrogen starvation, and YxATP was observed to increase from 80 mmol g-1 during the exponential growth phase to more than 130 mmol g-1 towards the end of the cultivation. The presented results illustrate the effect of nitrogen starvation on glycerol formation, protein turnover, catabolite inactivation of the sugar-transport system, the cellular composition, the cell cycle and growth energetics.

Ammonia↗

NMR study of chloride ion interactions with thylakoid membranes.

The role of Cl(-) in photosynthetic O(2) evolution has been investigated by observing the (35)Cl NMR linewidth under a variety of conditions in aqueous suspensions of chloroplasts, primarily for the halophytes Avicennia germinans, Avicennia marina, and Aster tripolium but also for spinach. The line broadening shows there is weak, ionic binding of Cl(-) to thylakoids, the bound Cl(-) exchanging rapidly (>>10(4) sec(-1)) with free Cl(-) in solution. The binding is necessary for O(2) evolution to occur. Michaelis-Menten constants obtained from the Cl(-) dependence of the O(2) evolution rate are approximately 15-70 mM for the halophytes compared with 0.6 mM for spinach (0.5 mM with Br(-)). There appear to be two types of Cl(-) binding sites in halophytes, of which the stronger is the activator, at lower [Cl(-)], of O(2) evolution. The (35)Cl line broadening includes a nonspecific interaction, which becomes apparent at high Cl(-) concentrations (>/=0.5 M).

Journal Article↗

Local extinction and the evolution of dispersal rates: causes and correlations.

We present the results of individual-based simulation experiments on the evolution of dispersal rates of organisms living in metapopulations. We find conflicting results regarding the relationship between local extinction rate and evolutionarily stable (ES) dispersal rate depending on which principal mechanism causes extinction: if extinction is caused by environmental catastrophes eradicating local populations, we observe a positive correlation between extinction and ES dispersal rate; if extinction is a consequence of stochastic local dynamics and environmental fluctuations, the correlation becomes ambiguous; and in cases where extinction is caused by dispersal mortality, a negative correlation between local extinction rate and ES dispersal rate emerges. We conclude that extinction rate, which both affects and is affected by dispersal rates, is not an ideal predictor for optimal dispersal rates.

Animals↗

The expression of alpha D-chains in the hemoglobin of adult ostrich (Struthio camelus) and American rhea (Rhea americana). The different evolution of adult bird alpha A-, alpha D- and beta-chains.

The hemoglobin of adult American rhea (Rhea americana) and ostrich (Struthio camelus) contains two components identified to be HbA (alpha 2A beta 2) and HbD (alpha 2D beta 2). The amino-acid sequence of alpha D-chains from HbD of adult American rhea and ostrich has been determined. The sequence was studied by Edman degradation of tryptic peptides and chemical cleavage products in a liquid phase sequencer. By homologous comparison with pheasant HbD (Phasianus colchicus colchicus), the alpha D-chains of American rhea differ by 28 amino-acid exchanges, the alpha D-chains of ostrich by 23 residues. These differences are higher than those observed for alpha A- as well as for beta-chains of HbA from the same species. The ratio of amino-acid exchanges for beta:alpha A:alpha D for American rhea and ostrich is found to be 1:5.5:6.5. At present the reason for the differences in evolution rates for the beta-, alpha A- and alpha D-chains of bird hemoglobins is still unclear.

Amino Acid Sequence↗