Cloning expression and characterization of a thermostable exopolygalacturonase from Thermotoga maritima

A gene encoding for a thermostable exopolygalacturonase (exo-PG) from hyperthermophilic Thermotoga maritima has been cloned into a T7 expression vector and expressed in Escherichia coli. The gene encoded a polypeptide of 454 residues with a molecular mass of 51,304 Da. The recombinant enzyme was purified to homogeneity by heat treatment and nickel affinity chromatography. The thermostable enzyme had maximum of hydrolytic activity for polygalacturonate at 95 degrees C, pH 6.0 and retains 90% of activity after heating at 90 degrees C for 5 h. Study of the catalytic activity of the exopolygalacturonase, investigated by means of 1H NMR spectroscopy revealed an inversion of configuration during hydrolysis of alpha-(1-->4)-galacturonic linkage.     

Exopolygalacturonate lyase from Thermotoga maritima: cloning,characterization and organic synthesis application

A new exopolygalacturonate lyase (Pel) gene of the hyperthermophilic bacterium Thermotoga maritima was cloned and overexpressed in Escherichia coli cells. A 42 kDa monomeric Pel was shown to undergo N-terminal processing by cleavage at a putative site between alanine and serine residues. The enzyme catalyzes selectively a beta-4,5 elimination at the third galacturonic unit from the reducing end of polygalacturonic acid by producing (4-deoxy-alpha-L-threo-hex-4-enopyranosyluronic acid)-(1-->4)-(alpha-D-galactopyranosyluronic acid)-(1-->4)-alpha-D-galactopyranuronic acid (3) with a 60% yield. The optimum activity of the enzyme was detected at pH 9.5 and T> or=95 degrees C. The highly thermostable enzyme constitutes a useful catalyst for a simplified synthesis of 4,5-unsaturated trigalacturonic acid 3, a trisaccharide which is extremely difficult to obtain via chemical synthesis.     

Intercropping <Emphasis Type="Italic">Caragana arborescens</Emphasis> with <Emphasis Type="Italic">Salix miyabeana</Emphasis> to Satisfy Nitrogen Demand and Maximize Growth

Willow shows great promise as a biomass crop and is now used worldwide. However, willow is a nutrient and water demanding plant that often requires the use of nitrogen (N) fertilizer to maximize growth on poor soils. The intercropping of Salix miyabeana with the atmospheric N₂-fixing Caragana arborescens on poor soils of the Canadian Prairies could provide a portion of the N demand of the willow. The main objectives were to: (1) determine the yield potential, N nutrition and water use efficiency (WUE) of willow and Caragana grown in pure and mixed plantations across a range of soil productivity and (2) assess the extent of atmospheric N₂-fixation by the Caragana within the first rotation in central Saskatchewan. We found large differences in willow yields, foliar N and WUE across the sites. The willow yields (1.24 to 15.6 t dry matter ha⁻¹ over 4 years) were low compared to northeastern North American values and reflect the short and dry summers of the region. The yields were positively correlated to foliar N (ranging between 14.3 and 32.4 mg g⁻¹), whereas higher WUE (expressed as δ¹³C) were not positively correlated to water availability but to higher yields. Caragana N₂-fixation (measured using ¹⁵N isotope dilution) was not active at the most productive site but up to 60% of the foliar N was of atmospheric origin at the two other sites. Willow growth increased with Caragana proportions at the least productive site, which is typical of the benefits of N₂-fixing plants on the growth of other plants on poor soils. At the most productive site, Caragana decreased the growth of willow early on due to competition for resources, but willow eventually shaded Caragana to a point of significant canopy decline and dieback. It is therefore more appropriate to intercrop the two species on less productive soils as Caragana is more likely to add N to the system via N₂-fixation and is less likely to be shaded out by willow.     

First Clarkforkian Equivalent Land Mammal Age in the Latest Paleocene Basal Sparnacian Facies of Europe: Fauna,Flora, Paleoenvironment and (Bio)stratigraphy

The Paleocene-Eocene Thermal Maximum (PETM) is correlated with the first occurrences of earliest modern mammals in the Northern Hemisphere. The latest Paleocene Clarkforkian North American Land Mammal Age, that has yielded rodents and carnivorans, is the only exception to this rule. However, until now no pre-PETM localities have yielded modern mammals in Europe or Asia. We report the first Clarkforkian equivalent Land Mammal Age in the latest Paleocene deposits of the basal Sparnacian facies at Rivecourt, in the north-central part of the Paris Basin. The new terrestrial vertebrate and macroflora assemblages are analyzed through a multidisciplinary study including sedimentologic, stratigraphic, isotopic, and palynological aspects in order to reconstruct the paleoenvironment and to evaluate biochronologic and paleogeographic implications. The mammals are moderately diverse and not abundant, contrary to turtles and champsosaurs. The macroflora is exceptional in preservation and diversity with numerous angiosperms represented by flowers, fruits, seeds and wood preserved as lignite material, revealing an abundance of Arecaceae, Betulaceae, Icacinaceae, Menispermaceae, Vitaceae and probably Cornaceae. Results indicate a Late Paleocene age based on carbon isotope data, palynology and vertebrate occurrences such as the choristoderan Champsosaurus, the arctocyonid Arctocyon, and the plesiadapid Plesiadapis tricuspidens. However, several mammal species compare better with the earliest Eocene. Among these, the particular louisinid Teilhardimys musculus, also recorded from the latest Paleocene of the Spanish Pyrenees, suggests a younger age than the typical MP6 reference level. Nevertheless, the most important aspect of the Rivecourt fauna is the presence of dental remains of a rodent and a “miacid” carnivoran, attesting to the presence of two modern mammalian orders in the latest Paleocene of Europe. Interestingly, these two groups are also the only modern groups recorded from the latest Paleocene of North America, making Rivecourt the first direct equivalent to the Clarkforkian Land Mammal Age outside of North America.     

Microbial Diversity During Cellulose Decomposition in Different Forest Stands: I. Microbial Communities and Environmental Conditions

We studied the effect of forest tree species on a community of decomposers that colonize cellulose strips. Both fungal and bacterial communities were targeted in a native forest dominated by beech and oak and 30-year-old beech and spruce plantations, growing in similar ecological conditions in the Breuil-Chenue experimental forest site in Morvan (France). Microbial ingrowths from the 3rd to 10th month of strip decomposition (May to December 2004) were studied. Community composition was assessed using temperature gradient gel electrophoresis with universal fungal (ITS1F, ITS2) and bacterial (1401r, 968f) primers. Soil temperature and moisture as well as fungal biomass were also measured to give additional information on decomposition processes. Changing the dominant tree species had no significant influence in the number of decomposer species. However, decomposer community composition was clearly different. If compared to the native forest, where community composition highly differed, young monocultures displayed similar species structure for fungi and bacteria. Both species numbers and community composition evolved during the decay process. Time effect was found to be more important than tree species. Nevertheless, the actual environmental conditions and seasonal effect seemed to be even more determining factors for the development of microbial communities. The course and correlations of the explored variables often differed between tree species, although certain general trends were identified. Fungal biomass was high in summer, despite that species richness (SR) decreased and conversely, that high SR did not necessarily mean high biomass values. It can be concluded that the growth and development of the microbiological communities that colonized a model material in situ depended on the combination of physical and biological factors acting collectively and interdependently at the forest soil microsite.     

Modern pollen rain in savanna and forest ecosystems of Gabon and Cameroon,Central Atlantic Africa

Eighty modern soil surface and litter samples from southern Cameroon and Gabon, Central Atlantic Africa (5°N–4°S, 10°–15°W), were analysed for pollen content. The samples are distributed among two main vegetation types: savanna (8 samples) and forest (71 samples). The aim of this study is to provide new data on the modern pollen rain in the Guineo-Congolian phytogeographical region, mainly in forest communities (secondary and mature forests on well drained soils, and hygrophilous forests) and to interpret these data using diagrams of pollen percentages and numerical analyses. The savannas are well identified by high frequencies of non-arboreal pollen with as pollen marker the Poaceae, and the forests by high frequencies of arboreal pollen with as important families the Burseraceae, Caesalpiniaceae, Mimosaceae, Euphorbiaceae and Sapindaceae. Within the forest ecosystem, secondary and mature forests on well drained soils can be differentiated on the basis of distinct assemblages of tree pollen taxa such as Zanthoxylum, Phyllanthus, Tetrorchidium, Margaritaria discoidea in secondary forest spectra and abundance of Burseraceae, Caesalpiniaceae, Sapindaceae in mature forest ones. In addition, hygrophilous forests are well identified by the presence of high pollen contributors such as Uapaca, Nauclea, Macaranga and Raphia. This work shows that the major vegetation communities occurring today in Cameroon and Gabon can be well differentiated by their pollen assemblages.     

Lead accumulation in the roots of grass pea (Lathyrus sativus L.): a novel plant for phytoremediation systems?

Eleven day-old grass pea plants (Lathyrus sativus L.) were grown hydroponically for 96 h in the presence of 0.5 mM lead nitrate (Pb(NO(3))(2)). The survival rate was 100%. The mean lead content (measured by ICP-OES) in root tissues was 153 mg Pb g(-1) dry matter. Over three quarters of the lead was not labile. Compared with control plants, lead-exposed plants showed a six-fold, two-fold and three and a half-fold reduction in their root calcium, zinc and copper contents, respectively. Together, these results suggested that Lathyrus sativus L. was tolerant to a deficiency in essential nutrients and able to store large amounts of lead in its root tissues. Therefore, it could be used for the development of new rhizofiltration systems.     

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 February 2013–31 March 2013

This article documents the addition of 142 microsatellite marker loci to the Molecular Ecology Resources database. Loci were developed for the following species: Agriophyllum squarrosum, Amazilia cyanocephala, Batillaria attramentaria, Fungal strain CTeY1 (Ascomycota), Gadopsis marmoratus, Juniperus phoenicea subsp. turbinata, Liriomyza sativae, Lupinus polyphyllus, Metschnikowia reukaufii, Puccinia striiformis and Xylocopa grisescens. These loci were cross‐tested on the following species: Amazilia beryllina, Amazilia candida, Amazilia rutila, Amazilia tzacatl, Amazilia violiceps, Amazilia yucatanensis, Campylopterus curvipennis, Cynanthus sordidus, Hylocharis leucotis, Juniperus brevifolia, Juniperus cedrus, Juniperus osteosperma, Juniperus oxycedrus, Juniperus thurifera, Liriomyza bryoniae, Liriomyza chinensis, Liriomyza huidobrensis and Liriomyza trifolii.     

Biology and ecology of sea catfish (Ariidae) of estuarine,lagoon and coastal ecosystems in West Africa

The family Ariidae, sea catfish of the order Siluriformes, is widely distributed throughout the world, particularly in tropical and subtropical areas. The three species of Ariidae found on the coasts and estuaries of West Africa are the smoothmouth catfish Carlarius heudelotii (Valenciennes 1840), the rough-head catfish Carlarius latiscutatus (Günther 1864) and the Guinean sea catfish Carlarius parkii (Günther 1864). They have been increasingly exploited by artisanal and industrial coastal fisheries in recent decades, but there is still little information available on their ecology and biology. The aim of this study was to deepen our knowledge of these three West African Ariidae species based on a dataset collected between 1980 and 2013 during experimental fishing programmes. They were carried out in Mauritania in the Banc d'Arguin National Park, in Senegal in the Sine Saloum estuary including the Bamboung Marine Protected Area (MPA), in The Gambia in the Gambia estuary, in Guinea-Bissau in the Urok Islands MPA in the Bijagos archipelago, in Guinea in the Fatala estuary and Dangara inlet, and in Côte d'Ivoire in the Ebrié Lagoon. C. latiscutatus accounted for 65%, C. parkii for 29% and C. heudelotii for 6% of total number of Ariidae sampled. C. latiscutatus was abundant in the Sine Saloum and Gambia estuaries as well as in Guinea and Guinea-Bissau and was the only species present in the Ebrié Lagoon. C. parkii was in the majority in Mauritania. The three species were recorded in a salinity range of 0 to 50, a temperature range of 19 to 34°C, in areas 1.7 to 15 m depth, and transparency ranging from 0.1 to 4 m (Secchi disk depth). C. heudelotii was present in less saline (25 vs. 32–34), less warm (27 vs. 29°C) and less transparent (0.8 m vs. 1.6 m) waters than the two other species. The maximum sizes (453 mm, 614 mm and 525 mm for, respectively, C. heudelotii, C. latiscutatus and C. parkii) were comparable to those recorded at sea. Length–weight relationships calculated for each species showed b coefficients greater than 3. Sex ratios were always in favour of females. The number of mature individuals and their smallest size at maturity were calculated per species, sex and study area. A size of 27–28 cm at first maturity was estimated for females of C. latiscutatus. A few dozen records made it possible to describe fecundity and cases of oral incubation by females. The diet of the three species was composed of crustaceans, fish and molluscs, confirming their classification as generalist predators. Thanks to their high environmental tolerance, these sea catfish populations are able to occupy both the continental shelf and adjacent estuaries throughout their life cycle, with the exception of spawning, which generally takes place at sea.