Effect of nutrient availability on hyphal maturation and topographical sensing in Aspergillus niger

Topographical sensing (thigmotropism) is an essential component of efficient fungal growth. It is an important element in the complex pathway of sensory and mechanical elements that drive and control the growing hyphal tip, a fuller understanding of which will bring the mycological community a step closer to complete comprehension of the hyphal growth mode. Previous work has led us to hypothesize that the stress induced by nutrient deficiency causes structural changes in the hyphal tip that induces a thigmotropic response in Aspergillus niger, a soil fungus that does not display thigmotropism under normal conditions. In this study, we have sought to identify some of the factors that influence this induction of thigmotropism using a novel combination of microengineered substrates and imaging and analysis techniques to quantify thigmotropic behavior in complex hyphal systems. We have shown that the sensitivity of fungal contour sensing appears to be directly linked to nutrient availability and hypothesize that this may be caused by a stress-induced flattening of the tip and increased immaturity of the hyphal apex. Parts of this paper were presented at the Mycological Society of Japan (MSJ)/British Mycological Society (BMS) Joint Symposium, “The new generation of mycologists in Japan and the UK” held in Chiba, Japan, on June 3, 2006.     

Inhibition of <Emphasis Type="Italic">Zizania latifolia</Emphasis> growth by <Emphasis Type="Italic">Phragmites australis</Emphasis>: an experimental study

The influence of Phragmites australis on the growth of Zizania latifolia, and the morphological characteristics of both species, were investigated at two nutrient levels. The experiment was carried out over three growing seasons between May 2003 and December 2005. The experimental plot was longitudinally divided into two equal halves, one of which was planted with Zizania latifolia (Zizania zone) and the other one with Phragmites australis (Phragmites zone). Four weeks after transplantation, the plot was again divided horizontally, perpendicular to the previous division, and subject to low (LN) and high nutrient (HN) treatments. Measured growth indices of the two species were more developed in the HN than in the LN treatments. Both species were observed to invade each other’s zone, but Phragmites latifolia seemed to out-compete Zizania latifolia individuals. This was evident from the decrease in below ground biomass of Zizania latifolia in the third growing season and deterioration in above ground organs with time. It was concluded that Phragmites australis out-competes Zizania latifolia due to better developed root and rhizome system.     

Nutrient status and retention in pristine and disturbed wetlands in Uganda: management implications

Wetlands in Uganda experience different forms of human pressure ranging from drainage for agriculture and industrial development to over harvesting of wetland products. In order to develop sustainable management tools for wetland ecosystems in Uganda and the Lake Victoria Region, water quality analyses were carried out in a rural undisturbed (pristine) wetland (Nabugabo wetland in Masaka) and two urban wetlands that are experiencing human and urban development pressure (the Nakivubo wetland in Kampala and Kirinya wetland in Jinja). The former wetland forms the main inflow into Lake Nabugabo while the other two border the northern shore of Lake Victoria, Uganda. Nabugabo wetland buffers Lake Nabugabo against surface runoff from the catchment, while Nakivubo and Kirinya wetlands provides a water treatment function for wastewater from Kampala City and Jinja town respectively, in addition to buffering Lake Victoria against surface runoff. Water quality was assessed in all the wetland sites, and in addition nutrient content and storage was investigated in the main plant species (papyrus, Phragmites, Miscanthidium and cocoyam) in Nakivubo and Kirinya wetlands. A pilot experiment was also carried out to assess the wastewater treatment potential of both the papyrus vegetation and an important agricultural crop Colocasia esculenta (cocoyam). Low electrical conductivity, ammonium–nitrogen and ortho-phosphate concentrations were recorded at the inflow into Nabugabo wetland (41.5 μS/cm; 0.91 mg/l and 0.42 mg/l respectively) compared to the Nakivubo and Kirinya wetlands (335 μS/cm; 31.68 mg/l and 2.83 mg/l and 502 μS/cm; 10 mg/l and 1.87 mg/l respectively). The papyrus vegetation had higher biomass in Nakivubo and Kirinya wetlands (6.7 kg DW m⁻²; 7.2 kg DW m⁻² respectively), followed by Phragmites (6.5, 6.7), cocoyams (6.4, 6.6) and Miscanthidium (4.0, 4.2). The papyrus vegetation also exhibited a higher wastewater treatment potential than the agricultural crop (cocoyam) during the pilot experiment (maximum removal degree of ammonium–nitrogen being 95% and 67% for papyrus and yams). It was concluded that urbanisation pressure reduces natural wetland functioning either through the discharge of wastewater effluent or the degradation of natural wetland vegetation. It is recommended that wetland vegetation be restored to enhance wetland ecosystem functioning and for wetlands that are not yet under agricultural pressure, efforts should be made to halt any future encroachment.     

Impact of grazing regime on a Mongolian forest steppe

Question: What is the impact of grazing regime on plant species abundance, plant growth form, plant productivity and plant nutrient concentrations in a forest steppe? Location: Hustai National Park in the forest steppe region of Mongolia. Methods: On the Stipa steppe we applied three different grazing regimes by using; (1) one type of exclosure which excluded grazing by large mammalian herbivores, mainly takh (Przewalski horse), (2) another type of exclosure that excluded both large and small (Siberian marmots) mammalian herbivores, and (3) control plots which were freely grazed. We measured species frequencies, tiller densities, plant biomass and nitrogen concentrations of the vegetation. Results: Exclusion from grazing by takh and marmots significantly increased plant standing crop, but marmot grazing and full grazing did not show significant differences. Protection from grazing decreased forage quality, shown by a lower N-concentration of the standing crop. However, this was solely the result of the lower live-dead ratio of the vegetation. The frequency of the rhizomatous Leymus chinensis decreased under reduced grazing, as did the frequency of the total of rhizomatous species. The frequency of Stipa krylovii increased under reduced grazing, as did its basal areas, tiller density and tussock height. Conclusion: Reduced grazing leads to a lower abundance of rhizomatous species and an increase in tussock species.     

Impact of sheep grazing on nutrient budgets of dry heathlands

Questions: What effect does sheep grazing have on the nutrient budgets of heathlands? Can grazing compensate for atmospheric nutrient loads in heathland ecosystems? What are the conclusions for heathland management? Location: Lüneburg Heath, NW Germany. Methods: During a one-year grazing experiment (stocking rate 1.1 sheep/ha) nutrient balances for N, Ca, K, Mg and P were calculated by quantifying input rates (atmospheric deposition, sheep excrement) and output rates (biomass removal, leaching). Results: Atmospheric nutrient deposition amounted to 22.8 kg.ha⁻¹.a⁻¹ for N and < 0.2 kg.ha⁻¹.a⁻¹ for P. Sheep excrement increased the inputs for N and P by ca. 3.5 and 0.2 kg.ha⁻¹.a⁻¹, respectively. Grazing reduced N- and P-stores in the above-ground biomass by 25.6 and 1.9 kg.ha⁻¹.a⁻¹, respectively. N-and P-losses via leaching amounted to 2.2 and < 0.2 kg.ha⁻¹.a⁻¹. Output:input ratios for P were high, indicating that grazing severely affected P-budgets of heaths. Conclusions: Our results suggest that sheep grazing has the potential to compensate for atmospheric nutrient loads (particularly for current N deposition rates). However, in the long term the combination of elevated N-deposition and P-loss due to grazing may cause a shift from N-(co-) limited to more P-(co-) limited plant growth. To counteract an aggravation of P-deficiency in the long term, grazing may be combined with management measures that affect P-budgets to a lesser extent (e.g. prescribed burning).     

Glucose transport by epithelia prepared from harvested enterocytes

Transformed and cultured cell lines have significant shortcomings for investigating the characteristics and responses of native villus enterocytes in situ. Interpretations of results from intact tissues are complicated by the presence of underlying tissues and the crypt compartment. We describe a simple, novel, and reproducible method for preparing functional epithelia using differentiated enterocytes harvested from the small intestine upper villus of adult mice and preterm pigs with and without necrotizing enterocolitis. Concentrative, rheogenic glucose uptake was used as an indicator of epithelial function and was demonstrated by cellular accumulation of tracer ¹⁴C d-glucose and Ussing chamber based short-circuit currents. Assessment of the epithelia by light and immunofluorescent microscopy revealed the harvested enterocytes remain differentiated and establish cell–cell connections to form polarized epithelia with distinct apical and basolateral domains. As with intact tissues, the epithelia exhibit glucose induced short-circuit currents that are increased by exposure to adenosine and adenosine 5′-monophosphate (AMP) and decreased by phloridzin to inhibit the apical glucose transporter SGLT-1. Similarly, accumulation of ¹⁴C d-glucose by the epithelia was inhibited by phloridzin, but not phloretin, and was stimulated by pre-exposure to AMP and adenosine, apparently by a microtubule-based mechanism that is disrupted by nocodazole, with the magnitudes of responses to adenosine, forskolin, and health status exceeding those we have measured using intact tissues. Our findings indicate that epithelia prepared from harvested enterocytes provide an alternative approach for comparative studies of the characteristics of nutrient transport by the upper villus epithelium and the responses to different conditions and stimuli.     

Combined arginine and ascorbic acid treatment induces apoptosis in the hepatoma cell line HA22T/VGH and changes in redox status involving the pentose phosphate pathway and reactive oxygen and nitrogen species

Arginine is a physiological substrate for nitric oxide synthase to generate nitric oxide (NO), which can influence tumor cell survival, while ascorbic acid is selectively toxic for cancer cells. This study explored the effect of an arginine/ascorbic acid combination on human cancer cell lines. The hepatoma cell line HA22T/VGH was the most sensitive of the tested cells to combination treatment. A combination of 5.74 mM of arginine and 0.57 mM of ascorbic acid induced HA22T/VGH cell death through apoptosis and an increase in levels of reactive oxygen species and NO, as well as its stable products NO₂⁻ and NO₃⁻. The combination also reduced the activity of glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and transaldolase in the pentose phosphate pathway, a major mechanism for producing NADPH, resulting in a marked decrease in intracellular NADPH levels. A dramatic decrease in intracellular glutathione (GSH) levels, a decrease in the mitochondrial membrane potential, ATP depletion and release of cytochrome c were also seen. Caspase-9 and caspase-3 were activated, apoptotic protein Bax expression increased and the expression of the anti-apoptotic proteins Bcl-2 and Bcl-xL decreased. These results suggest that this combination induced HA22T/VGH cell death by interfering with redox state regulation by a reduction in pentose phosphate pathway activity and increasing oxidative and nitrosative stress.