Mineral uptake in tobacco leaf discs during different developmental stages of shoot organogenesis

Relationships between mineral uptake and tobacco shoot organogenesis were investigated during three morphogenic phases: phase 1, days 0-10, pre-meristem formation; phase 2, days 10-20, meristem initiation and formation; and phase 3, days 20-35, growth and differentiation of induced meristems into leafy shoots. The mineral content of both shoot-forming (SF) and non-shoot-forming (NSF) media was examined over the 35-day culture period. Both SF and NSF explants rapidly consumed iron during phase 1. Nitrate uptake in SF explants was high and independent of explant growth during phases 1 and 2, but greatest and strongly correlated with growth during phase 3. Phosphorus uptake was highest in SF explants during phases 2 and 3, and correlated with explant growth. Uptake of potassium, calcium and sulphur was strongly associated with explant growth during phase 3 whereas magnesium uptake was only poorly correlated with growth. Results from this study indicate that particular minerals may have an important role in regulating development as well as generally supporting growth.

Upland streamwater nitrate dynamics across decadal to sub-daily timescales: a case study of Plynlimon, Wales

Streamwater nitrate dynamics in the River Hafren, Plynlimon, mid-Wales were investigated over decadal to sub-daily timescales using a range of statistical techniques. Long-term data were derived from weekly grab samples (1984–2010) and high-frequency data from 7-hourly samples (2007–2009) both measured at two sites: a headwater stream draining moorland and a downstream site below plantation forest. This study is one of the first to analyse upland streamwater nitrate dynamics across such a wide range of timescales and report on the principal mechanisms identified. The data analysis provided no clear evidence that the long-term decline in streamwater nitrate concentrations was related to a decline in atmospheric deposition alone, because nitrogen deposition first increased and then decreased during the study period. Increased streamwater temperature and denitrification may also have contributed to the decline in stream nitrate concentrations, the former through increased N uptake rates and the latter resultant from increased dissolved organic carbon concentrations. Strong seasonal cycles, with concentration minimums in the summer, were driven by seasonal flow minimums and seasonal biological activity enhancing nitrate uptake. Complex diurnal dynamics were observed, with seasonal changes in phase and amplitude of the cycling, and the diurnal dynamics were variable along the river. At the moorland site, a regular daily cycle, with minimum concentrations in the early afternoon, corresponding with peak air temperatures, indicated the importance of instream biological processing. At the downstream site, the diurnal dynamics were a composite signal, resultant from advection, dispersion and nitrate processing in the soils of the lower catchment. The diurnal streamwater nitrate dynamics were also affected by drought conditions. Enhanced diurnal cycling in Spring 2007 was attributed to increased nitrate availability in the post-drought period as well as low flow rates and high temperatures over this period. The combination of high-frequency short-term measurements and long-term monitoring provides a powerful tool for increasing understanding of the controls of element fluxes and concentrations in surface waters.

Rapid and slow modulation of nitrate reductase activity in the red macroalga Gracilaria chilensis (Gracilariales, Rhodophyta): influence of different nitrogen sources

Nitrate is one of the most important stimuli in nitrate reductase (NR) induction, while ammonium is usually an inhibitor. We evaluated the influence of nitrate, ammonium or urea as nitrogen sources on NR activity of the agarophyte Gracilaria chilensis. The addition of nitrate rapidly (2 min) induced NR activity, suggesting a fast post-translational regulation. In contrast, nitrate addition to starved algae stimulated rapid nitrate uptake without a concomitant induction of NR activity. These results show that in the absence of nitrate, NR activity is negatively affected, while the nitrate uptake system is active and ready to operate as soon as nitrate is available in the external medium, indicating that nitrate uptake and assimilation are differentially regulated. The addition of ammonium or urea as nitrogen sources stimulated NR activity after 24 h, different from that observed for other algae. However, a decrease in NR activity was observed after the third day under ammonium or urea. During the dark phase, G. chilensis NR activity was low when compared to the light phase. A light pulse of 15 min during the dark phase induced NR activity 1.5-fold suggesting also fast post-translational regulation. Nitrate reductase regulation by phosphorylation and dephosphorylation, and by protein synthesis and degradation, were evaluated using inhibitors. The results obtained for G. chilensis show a post-translational regulation as a rapid response mechanism by phosphorylation and dephosphorylation, and a slower mechanism by regulation of RNA synthesis coupled to de novo NR protein synthesis.