Tribute in memory of Jacques Breton (1942–2018)

Photosynthesis Research - Jacques Breton spent his 39 years of professional life at Saclay, a center of the French Atomic Energy Commission. He studied photosynthesis with various advanced...     

Effects of phenol in antioxidant metabolism in matrinxã, Brycon amazonicus (Teleostei; Characidae)

Parameters of the antioxidant defense systems of Brycon amazonicus (matrinx?--a neotropical fish) exposed to phenol for 96 h plus the recovery over 1 and 2 weeks were studied in erythrocytes and liver. Hematocrit increase was observed during phenol exposure and recovery for 1 week. Total superoxide dismutases (SOD), glutathione peroxidase (GPx) and reduced glutathione (GSH) did not change during phenol exposure. Erythrocyte glucose-6-phosphate dehydrogenase (G6PDH) increased during that period while catalase (CAT) activity decreased during phenol exposure and recovery for 2 weeks. In the liver, SOD and CAT did not change, whereas GPx increased in the first week of recovery and decreased after 2 weeks. A late response was observed for G6PDH activity which increased only at the second week. Ascorbate concentration in the brain decreased during phenol exposure and increased over recovery. From our results it appears that the oxidative stress was limited in matrinx? exposed to phenol, but seemed to occur during the recovery period.     

Integral role of the I′-helix in the function of the “inactive” C-terminal domain of catalase–peroxidase (KatG)

Catalase–peroxidases (KatGs) have two peroxidase-like domains. The N-terminal domain contains the heme-dependent, bifunctional active site. Though the C-terminal domain lacks the ability to bind heme or directly catalyze any reaction, it has been proposed to serve as a platform to direct the folding of the N-terminal domain. Toward such a purpose, its I′-helix is highly conserved and appears at the interface between the two domains. Single and multiple substitution variants targeting highly conserved residues of the I′-helix were generated for intact KatG as well as the stand-alone C-terminal domain (KatG^C). Single variants of intact KatG produced only subtle variations in spectroscopic and catalytic properties of the enzyme. However, the double and quadruple variants showed substantial increases in hexa-coordinate low-spin heme and diminished enzyme activity, similar to that observed for the N-terminal domain on its own (KatG^N). The analogous variants of KatG^C showed a much more profound loss of function as evaluated by their ability to return KatG^N to its active conformation. All of the single variants showed a substantial decrease in the rate and extent of KatG^N reactivation, but with two substitutions, KatG^C completely lost its capacity for the reactivation of KatG^N. These results suggest that the I′-helix is central to direct structural adjustments in the adjacent N-terminal domain and supports the hypothesis that the C-terminal domain serves as a platform to direct N-terminal domain conformation and bifunctionality.     

Effects of antimicrobial activity of silver nanoparticles on in vitro establishment of G × N15 (hybrid of almond × peach) rootstock

In the present investigation were evaluated the antifungal and antibacterial activities of Nano-silver (NS). Two separate experiments were done to evaluate the potential of silver nanoparticles in controlling the contamination of G × N15 micro-propagation. In the first experiment, a factorial experiment based on a completely randomized design with 15 treatments including five different NS concentrations (0, 50, 100, 150 and 200 ppm) and three soaking time of explants (3, 5 and 7 min) with five replications was conducted. In the other experiment, NS was added to Murashige and Skoog (MS) medium with concentrations of 0, 50, 100, 150 and 200 ppm in a completely randomized design. Results showed that the application of 100 and 150 ppm NS both as an immersion and as added directly to the culture medium significantly reduces internal and external contaminations compared with the control group. Using NS in culture medium was more efficient to reduce fungal and bacterial contamination than immersion. High concentrations of NS had an adverse effect on the viability of G × N15 single nodes and this effect was more serious in immersed explants in NS than directly added NS ones regarding the viability of buds and plantlet regeneration. In conclusion, due to high contamination of woody plants especially fruit trees and also adverse environmental effects of mercury chloride, the NS solution can be used as a low risk bactericide in micro-propagation of G × N15 and can be an appropriate alternative to mercury chloride in the future.     

Global diversity of oligochaetous clitellates (“Oligochaeta”; Clitellata) in freshwater

One of the predicted consequences of climate change is an increase in the occurrence of extreme rainfall and a subsequent increase in frequency of high flow events in rivers. High flow events have the potential to impact estuarine communities like mussel assemblages due to decreased salinity and/or the transport of sediments, organic matter and nutrients from the terrestrial environment to the estuary. The impact of two high flow events was investigated using mussels located within the Conwy estuary, North Wales, using the ‘Beyond BACI’ approach. Three study sites were chosen, the potentially impacted site (Conwy) and two control sites located outside the estuary. Sampling took place over 18 months with samples being collected before and after each event. On each sampling occasion, the following data were collected: the total haemocyte count (THC) and condition index (CI) of the mussels and the diversity (Hlog_e) of their associated macrofauna. A significant effect of the first event (22nd October 2004) was found on the CI of the Conwy mussels, whereas a significant effect of the second event (10th October 2005) was found on mussel THC. No effect of either event was found on the diversity of the associated fauna. The results of this study suggest that any increase in the number or intensity of heavy precipitation or high flow events have potential implications for the health and resilience of estuarine mussel populations. Guest editors: J. Davenport, G. Burnell, T. Cross, M. Emmerson, R. McAllen, R. Ramsay & E. Rogan Challenges to Marine Ecosystems     

Characteristics of photosynthesis in maize leaves (С<Subscript>4</Subscript> plants) upon changes in the level of illuminance and nitrate nutrition

We studied assimilation of ¹⁴СО₂ and distribution of ¹⁴С among the products of 3-min-long photosynthesis of maize (Zea mays L.) leaves. The day before the experiment, half of the plants were fertilized with Ca(NO₃)₂ (1 g/L of water) at a rate of 6 L/m². Five days before the experiment, some plants were shaded for adaptation (illuminance was reduced by 50%). On the day of the experiment (before the application of ¹⁴СО₂), several shaded plants were exposed to direct sunlight for 3 min, and some plants grown at full light (light plants) were shaded for 3 min (illuminance of 50%). Unfertilized plants adapted for 5 days to shading showed photosynthesis of 75.9% of control level (full light). If light plants were transferred to shading for 3 min, their photosynthesis decreased to 42.1%. In plants shaded for 5 days and then transferred to full light, photosynthesis in 3 min was 96.3% of control level. At full light, fertilization with nitrate boosted photosynthesis to 132.6% as compared with control material, but photosynthesis decreased to 43.5 and 65.4% of control level in plants shaded for 5 days and those shaded for 3 min, respectively. At the same time, the plants shaded for 5 days and then exposed for 3 min to full light restored photosynthesis to almost control level (95.5%). Analysis of ¹⁴С distribution among the products of 3-min-long photosynthesis showed that, the same as in C₃ plants, a decrease in illuminance (especially a sudden one) in maize reduced the ratio between labeled sucrose and hexoses and elevates incorporation of ¹⁴С into malate, which indicated that its consumption in bundle sheath cells was suppressed. A decrease in the ratio between labeled sucrose and hexoses became more pronounced under the influence of nitrates with this effect also occurring in transport products of photosynthesis (20 cm below ¹⁴С-providing leaf area). In plants fertilized with nitrates, radioactivity of sucrose (% of radioactivity of soluble compounds) decreased in all the types of illumination. When illuminance was suddenly reduced for 3 min, incorporation of ¹⁴С into sucrose was 21.5 against 51.2% in light plants, and radioactivity of aspartate and malate sharply rose to 13.7 and 26.1% (against 2.1 and 8.9% in control material). Incorporation of ¹⁴С into compounds of glycolate pathway was low (less than 2.5%), but it was somewhat greater in nitrate plants. We concluded that the same mechanism of interaction between stomatal apparatus of leaf epidermis, invertase of mesophyll apoplast, and photosynthetic metabolism of carbon with electron flux via electron transport chain in chloroplasts of bundle sheath cells, which governs the rate of photosynthesis and assimilate export from the leaf but is triggered by the extent of consumption in the bundle sheath cells of C₄ acids produced in the mesophyll operates in C₄ plants (the same as in C₃ plants).     

Microsatellite assessment of the genetic diversity in indigenous populations of curimba (<Emphasis Type="Italic">Prochilodus argenteus</Emphasis>) in the São Francisco river (Brazil)

The freshwater fish curimba (Prochilodus argenteus) is an endemic species of the São Francisco River basin, with great aquaculture potential. The construction of several hydroelectric power dams along the river, and the pulsed releases of water from these dams altered and degraded the native fish habitats, which led to the development of a hatchery-reared fingerling restocking program. A comprehensive genetic baseline evaluation of the indigenous populations of curimba and of a hatchery stock was done using microsatellite markers. Pairwise F _ST values, Bayesian analysis and the number of migrants per generation were used to illustrate the genetic structure of the curimba populations along almost 3000 km of the river, suggesting the existence of three units of conservation: one in the Upper, a second in the Middle and a third including part of the Middle, Submiddle and Lower stretches. We have analyzed the genetic contribution of hatchery stock releases in two stretches of the river using individual admixture coefficients; and the results showed that admixture makes a negligible contribution to indigenous recruitment. We discussed about supplementation programs for this river from the perspective of three units of conservation and the risks associated with using domesticated fish.     

Distribution and some traits of biology of <Emphasis Type="Italic">Lycodes tanakae</Emphasis> (Perciformes: Zoarcidae) in Primor’e waters (Sea of Japan)

With consideration of investigations made in 2004–2010, the spatial distribution of Lycodes tanakae, as well as some traits of its biology in Primor’e waters, is described. This species occurs all over the investigated region—from 42° to 49°N as explained by hydrological properties of bottom waters and by the bottom relief of this water area. In Promor’e waters, Lycodes tanakae occurs at depths of 87–1034 m. However, the majority of specimens (90%) prefers depths of 200–700 m. Thus, it can be attributed to the mesobathial ecological fish group. In spring and summer, juveniles of L. tanakae live at similar depths (200–350 m) where this species seems to spawn. In catches, L. tanakae is represented by specimens 11–90 cm in length, 0.1–4.9 kg in weight, and age from 1 to 10 years. The bulk of catches consists of specimens 40–70 cm long (62%), up to 2.2 kg in weight (90%), and of the age 4–7 years (72%). Sexual dimorphism in linear dimensions is not determined in L. tanakae. In summer, the ration of L. tanakae in Primor’e waters consists predominantly of cephalopods (on average, 59.4% of food weight). A noticeable part belongs to decapods (19.8%) and fish juveniles (16.6%). The smallest analyzed specimens consume small decapods and polychaetes. At the body length over 40 cm, L. tanakae pass over to predation. The value of the daily ration of Lycodes tanakae is, on average, 1.5% of the body weight.     

Evolution of mixed-linkage (1 -> 3, 1 -> 4)-β-D-glucan (MLG) and xyloglucan in Equisetum (horsetails) and other monilophytes

Background and Aims

Horsetails (Equisetopsida) diverged from other extant eusporangiate monilophytes in the Upper Palaeozoic. They are the only monilophytes known to contain the hemicellulose mixed-linkage (1 → 3, 1 → 4)-β-d-glucan (MLG), whereas all land plants possess xyloglucan. It has been reported that changes in cell-wall chemistry often accompanied major evolutionary steps. We explored changes in hemicelluloses occurring during Equisetum evolution.

Methods

Hemicellulose from numerous monilophytes was treated with lichenase and xyloglucan endoglucanase. Lichenase digests MLG to di-, tri- and tetrasaccharide repeat-units, resolvable by thin-layer chromatography.

Key Results

Among monilophytes, MLG was confined to horsetails. Our analyses support a basal trichotomy of extant horsetails: MLG was more abundant in subgenus Equisetum than in subgenus Hippochaete, and uniquely the sister group E. bogotense yielded almost solely the tetrasaccharide repeat-unit (G4G4G3G). Other species also gave the disaccharide, whereas the trisaccharide was consistently very scarce. Tetrasaccharide : disaccharide ratios varied interspecifically, but with no consistent difference between subgenera. Xyloglucan was scarce in Psilotum and subgenus Equisetum, but abundant in subgenus Hippochaete and in the eusporangiate ferns Marattia and Angiopteris; leptosporangiate ferns varied widely. All monilophytes shared a core pattern of xyloglucan repeat-units, major XEG products co-chromatographing on thin-layer chromatography with non-fucosylated hepta-, octa- and nonasaccharides and fucose-containing nona- and decasaccharides.

Conclusions

G4G4G3G is the ancestral repeat-unit of horsetail MLG. Horsetail evolution was accompanied by quantitative and qualitative modification of MLG; variation within subgenus Hippochaete suggests that the structure and biosynthesis of MLG is evolutionarily plastic. Xyloglucan quantity correlates negatively with abundance of other hemicelluloses; but qualitatively, all monilophyte xyloglucans conform to a core pattern of repeat-unit sizes.     

Kinetics of Histidine Dissociation From the Heme Fe(III) in N-fragment (residues 1–56) of Cytochrome <Emphasis Type="Italic">c</Emphasis>

We have here investigated the dissociation kinetics of the His side chains axially ligated to the heme-iron in the ferric (1–56 residues) N-fragment of horse cyt c. The ligand deligation induced by acidic pH-jump occurs as a biexponential process with different pre-exponential factors, consistent with a structural heterogeneity in solution and the presence of two differently coordinated species. In analogy with GuHCl-denatured cyt c, our data indicate the presence in solution of two ferric forms of the N-fragment characterized by bis-His coordination, as summarized in the following scheme: His18–Fe(III)–His26 \rightleftharpoons His18–Fe(III)–His33. We have found that the pre-exponential factors depend on the extent of the pH-jump. This may be correlated with the different pK_a values shown by His26 and His33; due to steric factors, His26 binds to the heme–Fe(III) less strongly than His33, as recently shown by studies on denatured cyt c. Interestingly, the two lifetimes are affected by temperature but not by the extent of the pH-jump. The lower pK_a for the deligation reaction required the use of an improved laser pH-jump setup, capable of inducing changes in H⁺ concentration as large as 1 mM after the end of the laser pulse. For the ferric N-fragment, close activation entropy values have been determined for the two histidines coordinated to the iron; this result significantly differs from that for GuHCl-denatured cyt c, where largely different values of activation entropy were calculated. This underlines the role played by the missing segment (residues 57–104) peptide chain in discriminating deligation of the nonnative His from the sixth coordination position of the metal.     

Cooperation of adenosine and prostaglandin E<Subscript>2</Subscript> (PGE<Subscript>2</Subscript>) in amplification of cAMP–PKA signaling and immunosuppression

INTRODUCTION: We hypothesize that adenosine and PGE(2) could have a complementary immunosuppressive effect that is mediated via common cAMP-PKA signaling. MATERIALS AND METHODS: To test this hypothesis, the effect of adenosine and PGE(2) on the cytotoxic activity and cytokine production of lymphokine activated killer (LAK) cells was investigated. RESULTS: PGE(2) and adenosine inhibited LAK cells cytotoxic activity and production of INF-gamma, GM-CSF and TNF-alpha. In combination they showed substantially higher inhibition than each modality used alone. Using agonists and antagonists specific for PGE(2) and adenosine receptors we found that cooperation of PGE(2) and adenosine in their inhibitory effects are mediated via EP(2) and A(2A) receptors, respectively. LAK cells have 35-fold higher expression of EP(2) than A(2A). Combined PGE(2) and adenosine treatment resulted in augmentation of cAMP production, PKA activity, CREB phosphorylation and inhibition of Akt phosphorylation. Wortmannin and LY294002 enhanced the suppressive effects of adenosine and PGE(2). In contrast, Rp-8-Br-cAMPS, an inhibitor of PKA type I blocked their immunosuppressive effects, suggesting that the inhibitory effects of PGE(2) and adenosine are mediated via common pathway with activation of cAMP-PKA and inhibition of Akt. CONCLUSION: In comparison to other immunosuppressive molecules (TGF-beta and IL-10), adenosine and PGE(2) are unique in their ability to inhibit the executive function of highly cytotoxic cells. High intratumor levels of adenosine and PGE(2) could protect tumor from immune-mediated destruction by inactivation of the tumor infiltrating functionally active immune cells.     

Induction of androgenetic development of the brook charr (Salvelinus fontinalis) × Arctic charr (Salvelinus alpinus) hybrids in eggs derived from the parental species

Failure of interspecific androgenesis between brook charr (Salvelinus fontinalis, Mitchill 1814) and Arctic charr (Salvelinus alpinus, L.) has been attributed to the conflict between the egg cytoplasm of one species and the sperm nucleus of the other species. To overcome this incompatibility, sperm derived from the brook charr × Arctic charr hybrid male was used to induce androgenetic development in eggs originating from the parental species as well as their hybrids. The eggs were subjected to 420 Gy of X-radiation to damage the maternal nuclear DNA and inseminated with untreated sperm. Haploid zygotes were exposed to high hydrostatic pressure shock (7000 psi for 4 min), which was applied 420 min after insemination to inhibit the first cell cleavage and recover the diploid state of the zygote. The androgenetic diploid offspring that hatched from the brook charr, the Arctic charr and the hybrids eggs had survival rates of 4.7 ± 0.6%, 1.2 ± 0.4% and 16.8 ± 0.5%, respectively. Drastic mortality among the hatched androgenetic individuals was observed within the first five months of rearing. Cytogenetic analysis of the androgenetic progenies exhibited residues of the irradiated maternal nuclear genome in the form of radiation-induced chromosome fragments in 47% of the specimens that were examined. Interactions between the egg cytoplasm and the sperm nucleus, the low quality of the gametes, the expression of homozygous paternal lethal alleles and the incomplete inactivation of the maternal chromosomes were identified as factors responsible for the large mortality among androgenetic embryos and hatchlings.     

Evaluation of clinical significance of с.2956G&gt;A (rs112287730) polymorphism in <Emphasis Type="Italic">FBN1</Emphasis> gene in Marfan syndrome

Marfan syndrome (MFS) is an autosomal dominant inherited systemic disorder of connective tissue with many clinical manifestations in the cardiovascular, skeletal, and ocular systems. MFS is caused by mutations in the fibrillin-1 (FBN1) gene. To date, about 2000 FBN1 pathogenic variants that cause MFS or related phenotypes have been described. The c.2956G>A, p.Ala986Thr substitution (exon 25) in the FBN1 gene is described in the SNP database as rs112287730 with allele frequency of 0.02%. Although numerous published data exist, the clinical significance of this variant is unknown. Some studies identify this substitution as probably a pathogenic mutation, and others, as a polymorphism. Among Russian Marfan patients, the heterozygous c.2956G>A substitution was identified in four probands; three of them had familial history. To determine the clinical significance of this substitution, a segregation analysis of DNA samples of affected and unaffected family members was conducted. In the first case, a segregation of the c.2956G>A substitution with the disease was observed in the family: this substitution was detected in the heterozygous state in the three affected members, but not in the one unaffected member. However, the opposite observation occurred in the second familial case: three affected members did not have the c.2956G>A substitution, whereas it was found in one unaffected member. In addition, the molecular-genetic analysis of 110 ethnically unrelated unexplored individuals was performed. The c.2956G>A substitution was identified in two of 220 examined chromosomes (allele frequency 0.9%). Thus, it was established that the c.2956G>A substitution appears to be a polymorphism (nonpathogenic variant) and cannot cause MFS.     

Notes on the biology of Trypoxylon (Trypargilum) opacum Brèthes (Hymenoptera; Crabronidae) in Southern Brazil.

The present study investigated the abundance, seasonality and various life-history traits of Trypoxylon (Trypargilum) opacum. Using trap-nests, 320 nests of T. opacum were collected in the Parque Municipal das Araucárias in Southern Brazil (25 degrees 23' 36" S and 51 degrees 27' 19" W) over a 3 year period. Nesting was more frequent during the warm season. Nests consisted of a linear series of 1 to 8 brood cells separated by mud partitions, usually followed by an empty vestibular cell and final-closure mud plug. Brood cells were most commonly provisioned with spiders of the family Araneidae. Sex-ratio was strongly female biased, 3.4:1 females:males. Natural enemies attacking nests T. opacum included chrysidids, ichneumonids, sarcophagids, bombyliids and ants.     

Altitudinal variation in ant–aphid mutualism in nitrogen transfer of oak (<Emphasis Type="Italic">Quercus liaotungensis</Emphasis>)

Ant–plant relationship is a model for the study of the ecology and evolution of interspecific interactions. In direct ant–plant mutualism (i.e., plants providing food or nesting places for ants, and ants protecting the plants in return) ants provide nutrients to plants. However, whether a similar mechanism exists in indirect ant–plant mutualism (i.e., an ant–aphid–plant system) remains unknown. In this work, we used the ¹⁵N stable isotope method to study altitudinal variations in the roles of ants in the nutrient transfer of oak (Quercus liaotungensis). Our work shows that ants deliver nitrogen in indirect ant–plant interactions, and that the effect of nutrient transfer differed significantly with altitude. Ants’ trophic level at high altitudes was significantly lower than that at low altitudes, indicating that the degree of ant–aphid mutualism was greater at high altitudes, which may be beneficial in nitrogen transfer. Our work suggests that ant–aphid mutualism might be context dependent, such that it affects nutrient transfer in the food web, and that this context dependency is an important factor that influences altitudinal variation in nutrient transfer.