Identification of Arabidopsis rat mutants

Limited knowledge currently exists regarding the roles of plant genes and proteins in the Agrobacterium tumefaciens-mediated transformation process. To understand the host contribution to transformation, we carried out root-based transformation assays to identify Arabidopsis mutants that are resistant to Agrobacterium transformation (rat mutants). To date, we have identified 126 rat mutants by screening libraries of T-DNA insertion mutants and by using various “reverse genetic” approaches. These mutants disrupt expression of genes of numerous categories, including chromatin structural and remodeling genes, and genes encoding proteins implicated in nuclear targeting, cell wall structure and metabolism, cytoskeleton structure and function, and signal transduction. Here, we present an update on the identification and characterization of these rat mutants.     

Reproductive measures and chick provisioning of Cory's Shearwater Calonectris diomedea borealis in the Azores

Capsule Timing of breeding influenced wing-length at fledging, and egg size may be an indicator of fledging weight and the amount of food received by chicks.

Aims To investigate chick growth, temporal patterns of chick food provisioning and the importance of indices of parental condition or quality, egg size and hatching date, to predict nestling body mass and wing-length at fledging, and compare breeding and chick feeding characteristics between colonies in the northeast Atlantic.

Methods A survey of Cory's Shearwater nests was carried out at Vila islet. A sample of 52 chicks, ringed and weighed at hatching, was selected to study chick growth and food provisioning.

Results Hatching success (51%) was much lower than fledging success (87%). Both hatching date and egg size contributed to explain wing-length at fledging, but hatching date, which was negatively correlated with wing-length at fledging, had the most important contribution (22%). There was some indication that egg size may explain variation in fledging weight and the amount of food received by chicks. Food delivery and feeding frequency of chicks varied throughout the chick development stage and three phases were distinguished: (1) 0–29 days, the highest feeding frequency values and a linear increase in food delivery; (2) 30–69 days, an oscillation in food delivery and medium feeding frequencies; (3) 70–90+ days, a sharp decrease in both food delivery and feeding frequency.

Conclusion Variation in food availability did not seem sufficient to override the overall importance of indices of parental quality in determining reproductive measures and chick provisioning. Breeding and feeding characteristics were similar between colonies in the northeast Atlantic, with variability in chick provisioning higher further south.     

On the altitudinal limits of birds on BasilC Peak (Bioko Island; Equatorial Guinea)

Pérez del Val, J. 2000. On the altitudinal limits of birds on Basile Peak (Bioko Island; Equatorial Guinea). Ostrich 71(1 & 2): 342.

The lower altitudinal limit of the montane avifauna in Africa has generally been set at 1 500- 800 m, corresponding to vegetation belts of the main mountain ranges of East Africa. However, much lower limits have been reported from the East Usambaras (Tanzania), Mount Cameroon, and Bioko Island. Basilé Peak is a volcano of 3 011 m which constitutes the northern half of Bioko Island. Below 400–900 m the forest has been logged and largely replaced by cocoa plantations, but the vegetation is relatively undisturbed at higher altitudes. Annual rainfall ranges between 2 000 mm and 3 500 mm. From January 1990 to July 1992 mist-nets were set at 13 standardised stations on Basilé Peak; 1 427 birds of 71 species were captured. Previous data from non-standardised mist-netting at 6 other localities, together with sight and sound records, published data, and 1 047 skins in the collections of Museo Naçional de Ciençias Naturales (Madrid), Estacion Biologica de Donaiia (Sevilla) and the Natural History Museum (Tring) were also used to compile distribution lists. From published botanical surveys, I recognised four vegetation zones: Lowland Rain Forest up to 800 m; Mountain Rain Forest at 80CL1 800 m; Araliaceus Forest 1 800–2 500 m; and Ericaceus Stands above 2 500 m. An analysis of similarity (using the index of Czechanowski) grouped 68 bird species into altitudinal classes at 200 m intervals. This suggests a division between lowland and montane birds at 800–1 000 m, broadly coinciding with the change between Lowland and Mountain Rain Forest. There does not seem to be a discrete bird community restricted to either of the upper zones (Araliaceus Forest or Ericaeus Stands), with only the Yellow Bishop Euplectes capensis restricted to the upper 500 m of the mountain. Most species which reach the summit are found in all vegetation zones, notably the endemic subspecies of the African Goshawk Accipiter tachiro lopezi. It is suggested that low temperatures, high rainfall, and in particular the seasonal distribution of rainfall, may explain the lower altitudinal limits of the montane avifauna on Bioko and on Mount Cameroon.     

Fine-root biomass and soil properties in a semideciduous and a lower montane rain forest in Panama

The distribution of root biomass and physical and chemical properties of the soils were studied in a semideciduous and in a lower montane rain forest in Panama. Roots and soil samples were taken by means of soil cores (25 cm deep) and divided into five, 5-cm deep sections. Soils were wet-sieved to retrieve the roots that were classified in four diameter classes: very fine roots (<1 mm), fine roots (1–2 mm), medium roots (2–5 mm) and coarse roots (5–50 mm). Soil samples were analyzed for organic carbon, total nitrogen, available phosphorus, exchangeable bases, cation exchange capacity, pH, aluminium and exchangeable acidity. Total root biomass measured with the soil corer (roots <50 mm in diameter) was not different between the forests (9.45 t ha^-1), while biomass of very fine roots was larger in the mountains (2.00 t ha^-1) than in the lowlands (1.44 t ha^-1). The soils in the semideciduous forest were low in available phosphorus, while in the mountains, soils had low pH, high exchangeable aluminium and exchangeable acidity, and low concentration of exchangeable bases. Phosphorus was in high concentration only in the first 5 cm of the soil. In both forests, there was an exponential reduction of root biomass with increasing depth, and most of the variation in the vertical distribution of roots less than 2 mm in diameter was explained by the concentration of nitrogen in the soils. The results of this study support the hypothesis that a large root biomass in montane forests is related to nutrients in low concentration and diluted in organic soils with high CEC and low bulk density, and that fine root biomass in tropical forests in inversely related to calcium availability but not a phosphorus as has been suggested for other forests.     

Growth Rate of the Microalga Tetraselmis suecica Changes the Biochemical Composition of Artemia Species

The impact of different microalgal semicontinuous cultures on growth and biochemical composition in the next link of the food chain was tested using the filter feeder Artemia species as a model. The marine microalga Tetraselmis suecica was cultured semicontinuously with renewal rates between 10% and 50% and used to feed Artemia. Microalgal cultures maintained with a low renewal rate that had biochemical composition similar to that of the stationary-phase cultures commonly used in aquaculture produced poor growth and survival and low food-conversion efficiency compared to cultures maintained with a high renewal rate. Changes in the renewal rate in microalgal cultures also resulted in important changes in the gross biochemical composition of the filter feeder. The gross biochemical composition of the Artemia resembled that of the microalgae used as food except for total lipid content. The percentage of protein in the organic fraction of Artemia increased from 45% to 65% of the organic weight with increasing renewal rates in the microalgal cultures, while the carbohydrate percentage decreased under the same conditions. Higher renewal rates resulted in higher lipid percentages in the microalga, but in Artemia the percentage of lipids decreased from 19% of the organic weight with a renewal rate of 10%, to 13% with a renewal rate of 50%. The percentage of all polyunsaturated fatty acids in Artemia, including 20:5n-3, increased slightly with increasing renewal rates in the microalgal cultures. Results emphasize the importance of controlling microalgal nutritional value for the success of aquaculture food chains in which filter feeders are involved. Received October 15, 2000; accepted December 29, 2000.     

The Psychedelic State Induced by Ayahuasca Modulates the Activity and Connectivity of the Default Mode Network

The experiences induced by psychedelics share a wide variety of subjective features, related to the complex changes in perception and cognition induced by this class of drugs. A remarkable increase in introspection is at the core of these altered states of consciousness. Self-oriented mental activity has been consistently linked to the Default Mode Network (DMN), a set of brain regions more active during rest than during the execution of a goal-directed task. Here we used fMRI technique to inspect the DMN during the psychedelic state induced by Ayahuasca in ten experienced subjects. Ayahuasca is a potion traditionally used by Amazonian Amerindians composed by a mixture of compounds that increase monoaminergic transmission. In particular, we examined whether Ayahuasca changes the activity and connectivity of the DMN and the connection between the DMN and the task-positive network (TPN). Ayahuasca caused a significant decrease in activity through most parts of the DMN, including its most consistent hubs: the Posterior Cingulate Cortex (PCC)/Precuneus and the medial Prefrontal Cortex (mPFC). Functional connectivity within the PCC/Precuneus decreased after Ayahuasca intake. No significant change was observed in the DMN-TPN orthogonality. Altogether, our results support the notion that the altered state of consciousness induced by Ayahuasca, like those induced by psilocybin (another serotonergic psychedelic), meditation and sleep, is linked to the modulation of the activity and the connectivity of the DMN.     

A yeast protein analogous to Escherichia coli RecA protein whose cellular level is enhanced after UV irradiation

Summary In Saccharomyces cerevisiae, a protein was recognized by polyclonal antibodies raised against homogeneous Escherichia coli K12 RecA protein. The cellular level of the yeast protein called RecAsc (molecular weight 44 kDa, pI 6.3), was transiently enhanced after UV irradiation. Protease inhibitors were required to minimize degradation of the RecAsc protein during cell lysis. The RecAsc protein exhibited similar basal levels and similar kinetics of increase after UV irradiation in DNA-repair proficient (RAD ⁺) strains carrying mitochondrial DNA or not (rho ⁰). This was also true for the following DNA-repair deficient (rad ^-) strains: rad2-6 rad6-1 rad52-1, a triple mutant blocked in three major repair pathways; rad6-, a mutant containing an integrative deletion in a gene playing a central role in mutagenesis; pso2-1, a mutant that exhibits a reduced rate of mutagenesis and recombination after exposure to DNA cross-linking agents.     

Profiling the Bordetella pertussis proteome during iron starvation

Regulation of gene expression in response to local iron concentration is commonly observed in bacterial pathogens that face this nutrient limitation during host infection. In this study, a proteomic approach was used to analyze the differential protein expression of Bordetella pertussis under iron limitation. Whole cell lysates (WCL) and outer membrane fractions of bacteria grown either under iron-starvation or iron-excess conditions were analyzed by two-dimensional (2-D) gel electrophoresis. Statistical analysis revealed 36 proteins displaying differential expression, 9 with higher expression under iron-excess and 27 with increased expression under iron-starvation. These proteins were subjected to tryptic digestion and MALDI-TOF MS. Apart from those previously reported, we identified new low-iron-induced proteins that might help to explain the increased virulence of this phenotype. Additionally, we found evidence that at least one of the identified proteins, solely expressed under iron starvation, is highly immunogenic in infected individuals.     

Structure of a protein–detergent complex: the balance between detergent cohesion and binding

Despite the major interest in membrane proteins at functional, genomic, and therapeutic levels, their biochemical and structural study remains challenging, as they require, among other things, solubilization in detergent micelles. The complexity of this task derives from the dependence of membrane protein structure on their anisotropic environment, influenced by a delicate balance between many different physicochemical properties. To study such properties in a small protein–detergent complex, we used fluorescence measurements and molecular dynamics (MD) simulations on the transmembrane part of glycophorin A (GpAtm) solubilized in micelles of dihexanoylphosphatidylcholine (DHPC) detergent. Fluorescence measurements show that DHPC has limited ability to solubilize the peptide, while MD provides a possible molecular explanation for this. We observe that the detergent molecules are balanced between two different types of interactions: cohesive interactions between detergent molecules that hold the micelle together, and adhesive interactions with the peptide. While the cohesive interactions are detergent mediated, the adhesion to the peptide depends on the specific interactions between the hydrophobic parts of the detergent and the topography of the peptide dictated by the amino acids. The balance between these two parameters results in a certain frustration of the system and rather slow equilibration. These observations suggest how molecular properties of detergents could influence membrane protein stabilization and solubilization.     

The emergence of neural activity and its role in the development of the enteric nervous system

The enteric nervous system (ENS) is a vital part of the autonomic nervous system that regulates many gastrointestinal functions, including motility and secretion. All neurons and glia of the ENS arise from neural crest-derived cells that migrate into the gastrointestinal tract during embryonic development. It has been known for many years that a subpopulation of the enteric neural crest-derived cells expresses pan-neuronal markers at early stages of ENS development. Recent studies have demonstrated that some enteric neurons exhibit electrical activity from as early as E11.5 in the mouse, with further maturation of activity during embryonic and postnatal development. This article discusses the maturation of electrophysiological and morphological properties of enteric neurons, the formation of synapses and synaptic activity, and the influence of neural activity on ENS development.