Identification and spatial arrangement of high molecular weight proteins (Mr 300 000-330 000) co-assembling with microtubules from a cultured cell line (rat glioma C6)

The distribution of three high molecular weight proteins, MAP-1 (Mr 330 000), MAP-2 (Mr 300 000) and plectin (Mr 300 000) in various fractions obtained in cycles of temperature-dependent polymerization/depolymerization of microtubules from rat glioma C6 cells was studied. Using gel electrophoresis and immunoautoradiography/immunoblotting all three proteins were found to codistribute only partially with tubulin because considerable parts remained in the cold-insoluble fractions. Moreover, the proteins, particularly MAPs, were proteolytically degraded during cycling. By contrast, when microtubules were polymerized with taxol after isotonic cell lysis a considerable enrichment of MAP-1 and MAP-2 was achieved; again, plectin co-distributed only partially. In this procedure too, MAPs, especially MAP-2, were found to be highly subject to proteolysis, unless free Ca2+-ions were rigorously avoided. Proteolytic fragments generated from MAP-2 were of similar size independent of whether temperature- or taxol-dependent polymerization procedures were used, suggesting the occurrence of a MAP-2-specific protease. When the spatial arrangement of the high Mr proteins on taxol-polymerized C6 cell microtubules was directly visualized using gold-immunoelectron microscopy, a periodical, apparently helical, decoration of microtubules was found for MAP-1 and MAP-2; plectin was irregularly arrayed. A predominantly helical arrangement of both MAPs was demonstrated also for microtubules reconstituted from mammalian brain.     

Micropropagation and pathogen elimination in elderberry (Sambucus nigra L.)

Plant Cell, Tissue and Organ Culture (PCTOC) - To date, more than 75% of Austrian elderberry acreage is planted with cultivar ‘Haschberg’. This leads to high costs during harvest, since...     

Leaf Litter Decomposition of Three Riparian Tree Species and Associated Macroinvertebrates of Eswathu Oya,a Low Order Tropical Stream in Sri Lanka

Leaf decomposition, an important component of the organic matter dynamics in streams, has been widely examined in temperate regions but much less documented in tropical regions. We report here the first study of leaf decomposition in a Sri Lankan stream. The litterbag technique was used. Coarse (8 mm) and fine (100 µm) litterbags, that included or excluded macroinvertebrates respectively, were used to enclose leaves of three dominant riparian tree species: native Ochlandra stridula (bamboo), and the introduced Alstonia macrophylla and Hevea brasiliensis (rubber). A fourth set of litterbags contained a mixture of these three species. Leaf colonization by macroinvertebrates was highest in the early stages of the decomposition process on Hevea leaves but invertebrate densities declined later. The opposite colonization effect was observed on the native Ochlandra leaves: slow colonization with continuing low densities from the beginning to the end of the process. Decomposition of all three species was significantly faster in the coarse than in the fine mesh bags. Alstonia, which has the softest leaf tissue, was most rapidly decomposed while Ochlandra, with its tough structure, was the slowest. Among the invertebrates, insects were the most important leaf colonizing animals, with Diptera, Coleoptera and Trichoptera the most dominant. The invertebrate variety in the mixed bags was higher than in the single‐species leaf bags, where Chironomidae dominated the colonizing assemblages. This study has shown that toughness, indicated by the ‘specific weight of leaf tissue’, and the quality of the leaves was more important in determining breakdown rates than their origin. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Influence of anthropogenic activities on water quality of a tropical stream ecosystem

Water samples were collected from three sites located in the middle reach of the Njoro River, Kenya, and analysed for total phosphorus (TP), orthophosphate, ammonia‐nitrogen, and nitrate‐nitrogen to evaluate stressor sources (e.g. factories and wastewater ponds) and the general stream water quality. The stream surface water was also analysed for biochemical oxygen demand (BOD₅) to provide an overview of organic matter loading. Mugo, Egerton Bridge and the canning factory sites of the Njoro River had low water quality which is likely to be due to poor farming, partially treated effluents and poor provision of sanitation facilities to the riparian communities. The concentrations of the selected nutrients did not differ significantly among the three sites, presumably due to pollution of the whole stream reach by the catchment nutrient sources. High phosphate concentrations (i.e. ～0.76 mgPO₄ l^?1 and ～0.87 mgTP l^?1) at Canning Factory were recorded during the low flow dry season. Nitrate‐nitrogen concentrations varied significantly with water discharge which explained between 63 and 87% of the nutrient variability in the three sites. BOD₅ differed significantly among the three sites, with historical effects of wastewater and factory effluent discharge being reflected in the results of Egerton Bridge and Canning Factory. The concentrations of ammonia‐nitrogen, TP and orthophosphate were higher in the wastewater than in the river water whereas nitrate‐nitrogen was lower. This study indicates that the Njoro River is stressed by nutrients from the activities within its catchment. With the increasing population, the nutrient load to the river will continue to increase and the water quality will continue to deteriorate. Reductions of nutrient loads into the river as well as provision of sanitation facilities to the riparian communities are needed to control further water degradation.     

Leaf litter transport and retention in a tropical Rift Valley stream: an experimental approach

The impact of a drought on freshwater snail and trematode communities was investigated in a lake. Before the drought, 15 gastropod species (Valvatidae, Planorbidae, Lymnaeidae, Ancylidae, Physidae) and 10 trematode species (cercariaeum, xiphidiocercariae, echinostome, furcocercariae, notocotyle, lophocercous) were recorded. The rate of parasitism was 5.13% and there were 11 host species. The 2 major consequences of desiccation were the disappearance of snails, except Valvata piscinalis and Lymnaea peregra, and the absence of trematodes infecting the surviving snails. As soon as favourable conditions were restored, the littoral area was recolonized, first by hygrophilic and amphibious species, second by aquatic species. Nine months after the drought, the gastropod community was restored. Recolonization by the trematodes was delayed compared with that of gastropods. During the study, the overall prevalence was equal to 0.36% and only 4 trematode species and 5 host species were recorded. Because of the great variability of freshwater ecosystems, long-term studies are necessary to understand the dynamics of snail and trematode populations and determine the regulatory effect of parasitism in the field.     

Cytoskeleton-associated plectin: in situ localization, in vitro reconstitution, and binding to immobilized intermediate filament proteins

The association and interaction of plectin (Mr 300,000) with intermediate filaments and filament subunit proteins were studied. Immunoelectron microscopy of whole mount cytoskeletons from various cultured cell lines (rat glioma C6, mouse BALB/c 3T3, and Chinese hamster ovary) and quick-frozen, deep-etched replicas of Triton X-100-extracted rat embryo fibroblast cells revealed that plectin was primarily located at junction sites and branching points of intermediate filaments. These results were corroborated by in vitro recombination studies using vimentin and plectin purified from C6 cells. Filaments assembled from mixtures of both proteins were extensively crosslinked by oligomeric plectin structures, as demonstrated by electron microscopy of negatively stained and rotary-shadowed specimens as well as by immunoelectron microscopy; the binding of plectin structures on the surface of filaments and cross-link formation occurred without apparent periodicity. Plectin's cross-linking of reconstituted filaments was also shown by ultracentrifugation experiments. As revealed by the rotary-shadowing technique, filament-bound plectin structures were oligomeric and predominantly consisted of a central globular core region of 30-50 nm with extending filaments or filamentous loops. Solid-phase binding to proteolytically degraded vimentin fragments suggested that plectin interacts with the helical rod domain of vimentin, a highly conserved structural element of all intermediate filament proteins. Accordingly, plectin was found to bind to the glial fibrillar acidic protein, the three neurofilament polypeptides, and skin keratins. These results suggest that plectin is a cross-linker of vimentin filaments and possibly also of other intermediate filament types.     

Decomposition of leaf litter of Dombeya goetzenii in the Njoro River, Kenya

Decomposition of the leaves of Dombeya goetzenii (K. Schum) in the Njoro River is described and analysed. The loss of the ash-free dry mass was rapid during the first 14 d of exposure in the wet and humid zones. The leaves in the litter bags in the humid and wet zones were processed at a rate (±SD) of 0.005±0.001 d^-1 and 0.021±0.001 d^-1, respectively. The processing rates of the leaves in the wet zone differed significantly from those observed in the humid zone (t-value, p<0.05). The interchanged litter bags (i.e. from wet to humid zones vice versa) showed that the processing rates of the leaves in the litter bags interchanged from the wet zone to the humid zone was about 200 times lower than that of the leaves retained in the wet zone throughout; fourfold higher in the leaves in the litter bags which were transferred to the wet zone than in those leaves of the litter bags which were retained in the humid zone throughout the experiment. It took approximately 38 months for 90% of the leaf dry mass to be processed in the humid zone whilst it took 4 months for a similar percentage to be processed in the wet zone. It is concluded that the immersion and emersion of leaf litter, which may occur in the wet and humid zones, respectively, are important aspects of the decomposition process which may influence the quantity of nutrients in stream ecosystems.