Production of solamargine by in vitro cultures of Solanum paludosum

Callus cultures of Solanum paludosum were established from roots, hypocotyles, cotyledons and leaf limbs of plantlets cultivated in sterile conditions on a Murashige and Skoog's modified medium. Non organogenous calluses were obtained with addition of BA or kinetin (10^-5M to 10^-6M) as the cytokinin and 2,4-d or NAA (10^-5M to 10^-6M) as the auxin. These calluses permitted the establishment of a cell suspension culture with BA (10-6M) and 2,4-d (10^-6M). Zeatin (10^-6M) with IAA (10^-6M) gave rise to organogenous calluses. These organogenous callus cultures developed multiple shoots which either proliferated if they were cultivated on a medium containing zeatin with IAA or IBA or were able to regenerate into whole plants when zeatin was used as the only hormone. The different plant material produced solamargine, the main steroidal glycoalkaloid present in the unripe fruits. The best production was obtained with the fruits of regenerated plants from organogenous callus cultures after reintroduction of these plants in their brasilian biotope. The solamargine content of the two types of plant materials was about 0.06% and 2.5% (dry weight) respectively for the callus cultures and the fruits from in vitro plants. The fruits were harvested a year after the beginning of the plantlet regeneration step.Abbreviations HPTLC high performance thin layer chromatography - HPLC high performance liquid chromatography - 2,4-d 2,4-dichlorophenoxyacetic acid - BA benzylaminopurine - IAA 3-indolebutyric acid - NAA -naphthaleneacetic acid - IBA 3-indolebutyric acid - IPA isopentenyladenine     

Ecological and genetic differentiation of Barbus callensis populations in Tunisia

Enzyme electrophoresis on horizontal starch gel was carried out on 356 barbel individuals. The sampling comprised 278 individuals of the species Barbus callensis from 10 rivers in Tunisia. The other individuals belonged to reference species (outgroups) from France and Morocco. An ecological study was also carried out on Tunisian rivers. The results show a clear differentiation of the two samples from northwestern Tunisia, which was only partly correlated with ecological characteristics of the rivers they inhabit. There is no genetic cline, but rather a discontinuity between populations in the northwestern-most watershed and all the other Tunisian populations. This differentiation probably has a paleohistoric origin not only related to adaptation to ecological conditions but also to difficulties in colonizing the watersheds. The results do not indicate clearly a colonization direction for the genus Barbus in North Africa. Analysis of the Algerian populations would appear to be indispensable. Lastly, in contrast with the usual taxonomy, Morocco and Tunisia are populated by two closely related species, but B. callensis should remain the name of the Tunisian species, which was the first to be described in the small El Kebir basin, a river that flows from Tunisia to Algeria.     

Tolerance to osmotic shocks in rat kidney cortex and medulla

Kidney medulla cells of mammals have to cope with large changes in environmental osmolarity, a challenge most other mammalian cells never have to experience. In these last cells, application of osmotic shocks induces dramatic modifications in chromatin organization. The present paper reports on the changes of medulla cell chromatin in situ, in rat kidney slices submitted to osmotic challenges and in vitro, on preparations of extracted chromatin submitted to changes in environmental ion concentrations. Our results show that the chromatin of kidney medulla cells: (1) does not behave differently from the other mammalian chromatins when submitted in situ or in vitro to osmotic challenges; (2) presents in vitro physico-chemical characteristics similar to those of the other mammalian chromatins; and (3) is protected in vitro, as the other mammalian chromatins, from the disrupting effects of increases in inorganic ion concentrations by different compensatory organic solutes. The ability of kidney medulla cells to adapt to large increases in osmolarity could thus be related to a rapid control of the level of such compounds rather than to some rather specific, intrinsic molecular adaptations of macromolecules.     

Accumulation of cadmium, lead and strontium, and a role of calcium oxalate in water hyacinth tolerance

Eichhornia crassipes plants brought from the River Nile were cultured in jars containing river water supplemented with various concentrations of Cd, Pb, and Sr (0 to 100 μg cm-3), added simultaneously. Treatment continued for 20 d during which each cultivation solution was being replaced with fresh one every 3 d. The growth of Eichhornia was drastically retarded at heavy metal concentrations higher than 15 μg cm-3. At concentrations 15 or 25 μg cm-3, the accumulation of Cd and Pb to levels several times higher than those in control plants was found. More than 50 % of the uptaken metals were retained by roots alone. Leaves and leaf petiols received around 30 and 20 % of the accumulated metals, respectively. X-ray microanalysis indicated the presence of the three heavy metals in Ca oxalate crystals. Content of metals in the crystals increased progressively over time of exposure in a way similar to those in whole plant tissues. These results suggest a possible role for Ca oxalate crystalization in toxic heavy metal deposition and thus tolerance by Eichhornia. This revised version was published online in August 2006 with corrections to the Cover Date.     

Purification, characterization and molecular modelling of two toxin-like proteins from the Androctonus australis Hector venom.

Two toxin-like proteins (AahTL1 and AahTL3) were purified from the venom of the scorpion Androctonus australis Hector (Aah). AahTL1 and AahTL3 are the first non toxic proteins cross-reacting with AahI toxins group which indicates that these proteins can be used as a model of vaccins. In order to study structure-function relationships, their complete amino-acid sequences (66 residues) were determined, by automated Edman degradation. They show more than 50% of similarity with both AahI and AahIII antimammal toxins. Three-dimensional structural models of AahTL1 and AahTL3 constructed by homology suggest that the two proteins are structurally similar to antimammal scorpion alpha-toxins specific to voltage dependent Na+ channels. The models showed also that amino-acid changes between potent Aah toxins and both AahTL1 and AahTL3 disrupt the electrostatic potential gradient at their surface preventing their interaction with the receptor, which may explain their non toxicity.     

The invasive brown seaweed Sargassum muticum as new resource for alginate in Morocco: Spectroscopic and rheological characterization

The Japanese brown seaweed Sargassum muticum, recently invaded several shorelines worldwide including the Atlantic coast of Morocco with large well‐established populations. Within the framework of a sustainable strategy to control this invasive seaweed, we report on extraction yield, spectroscopic characterization and rheological properties of alginate, a commercially valuable colloid, from harvested biomass of S. muticum. Extraction yield was about 25.6% on dry weight basis. Infrared spectroscopy analysis shows that the obtained Fourier transform infrared spectra of the extracted biopolymer exhibit strong similarities with that of the commercial alginate. Furthermore, Proton nuclear magnetic resonance spectroscopy revealed that S. muticum alginate has almost equal amounts of β‐D‐mannuronic acid (M; 49%) and α‐L‐guluronic acid (G; 51%) with an M/G ratio of 1.04 and a high content of heteropolymeric MG GM diads suggesting a sequence distribution of an alternated polymer type. Rheological measurements were performed at different sodium alginate concentrations, temperatures and shear rates. The hydrocolloid exhibited pseudoplastic behavior and showed shear thinning, particularly at high solution concentration and low temperature which is consistent with the rheological behavior reported for commercial alginates. Considering the abundance of S. muticum in the Northwestern Atlantic coast of Morocco and the quality of the extracted hydrogel, this invasive species could be considered as a potential source of alginates.     

Role of neuromediators in the functioning of the human microbiota: “Business talks” among microorganisms and the microbiota-host dialogue

Current concepts concerning the social behavior of microorganisms inhabiting the human gastrointestinal (GI) tract and their role in the formation of integrated supracellular structures and in intercellular communication in the host–microbiota system are reviewed. The analysis of the literature data and of the results obtained by the authors indicates an important role of neuromediators (biogenic amines, amino acids, peptides, and nitric oxide) in intra- and interspecies microbial communication, as well as in the microbiota–host dialogue. The role of this dialogue for human health, its effect on the human psyche and social behavior, and the possibility of construction of probiotic preparations with a target-oriented neurochemical effect are discussed.     

Cleistogamy and phylogenetic position of Viola uliginosa (Violaceae) re‐examined

Viola uliginosa is an endangered species throughout its range in Central Europe. Based on an unusual combination of morphological characters, including a reported lack of cleistogamous (CL) flowers, it was previously placed in the monotypic subsection Repentes, section Viola. Available individual phylogenetic analyses identify V. uliginosa as a lineage deeply embedded in section Viola, but hitherto its exact position was unknown. Here we confirm the occurrence of cleistogamy in V. uliginosa. Unlike other species of section Viola, V. uliginosa produced both chasmogamous and CL flowers at the same time, not at different times of the year (seasonal cleistogamy). The two flower types differed conspicuously in morphology and microstructure, especially in size and shape of the pistil and stamens as confirmed by scanning electron microscopy. Secondly, we resolved the phylogenetic position of V. uliginosa as a lineage deeply embedded in subsection Rostratae by means of a multigene multispecies coalescent analysis using *BEAST. In this analysis, V. mirabilis and V. uliginosa were successive sisters to the rest of subsection Rostratae and, because allopolyploids exist between V. mirabilis and the last one, subsection Rostratae needs to comprise all these taxa to be monophyletic. The evolution of key morphological traits is briefly discussed.     

Toxic gain of function from mutant FUS protein is crucial to trigger cell autonomous motor neuron loss

FUS is an RNA‐binding protein involved in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Cytoplasmic FUS‐containing aggregates are often associated with concomitant loss of nuclear FUS. Whether loss of nuclear FUS function, gain of a cytoplasmic function, or a combination of both lead to neurodegeneration remains elusive. To address this question, we generated knockin mice expressing mislocalized cytoplasmic FUS and complete FUS knockout mice. Both mouse models display similar perinatal lethality with respiratory insufficiency, reduced body weight and length, and largely similar alterations in gene expression and mRNA splicing patterns, indicating that mislocalized FUS results in loss of its normal function. However, FUS knockin mice, but not FUS knockout mice, display reduced motor neuron numbers at birth, associated with enhanced motor neuron apoptosis, which can be rescued by cell‐specific CRE‐mediated expression of wild‐type FUS within motor neurons. Together, our findings indicate that cytoplasmic FUS mislocalization not only leads to nuclear loss of function, but also triggers motor neuron death through a toxic gain of function within motor neurons.