Fagus sylvatica (beech) dominates the montane forests of the Apennines and builds old-growth high-conservation value stands. However, recent severe drought-induced diebacks raise concern on the future persistence of these forests and of Southern European mesophilous woodlands overall, growing at their dry edge. To explore the history of Apennine beech-dominated forests, we draw on the multiproxy paleoecological record from Lago Verdarolo, which includes a robust vegetation-independent temperature reconstruction. Numerical techniques are used to investigate the drivers of long-term Mediterranean mountain forest dynamics. Specifically, we focus on disentangling the ecological factors that caused the shift from high-diversity mixed forests to beech-dominated stands and on assessing the occurrence of legacy effects on present-day forests. Abrupt climate change largely drove vegetation dynamics during the Late Glacial and Early Holocene. Species-rich mixed Abies alba (silver fir) forests dominated about 10,500—5500 years ago, under rather dry and warmer-than-today conditions (+ 1—2 °C) and limited fire occurrence. Cooler and moister summers and increasing fire activity caused declines in several fire-sensitive temperate deciduous trees (for example, Ulmus, Tilia, Fraxinus) and favored the establishment of fir-beech forests around 5500 years ago. Further enhancement of fire activity and farming around 2000 years ago led to local Abies alba extinction and forest impoverishment. We conclude that the currently widespread monospecific Apennine beech forests are the result of multi-millennial land-use intensification superimposed on Late Holocene cooling and moistening. Given their higher drought-tolerance compared to beech stands, reviving ancient species-rich mixed fir forests represents a feasible and ‘tested’ possibility to adapt forests to climate change.
For Trypanosoma brucei arginine and lysine are essential amino acids and therefore have to be imported from the host. Heterologous expression in Saccharomyces cerevisiae mutants identified cationic amino acid transporters among members of the T. brucei AAAP (amino acid/auxin permease) family. TbAAT5-3 showed high affinity arginine uptake (Km 3.6 ± 0.4 μM) and high selectivity for L-arginine. L-arginine transport was reduced by a 10-times excess of L-arginine, homo-arginine, canavanine or arginine-β-naphthylamide, while lysine was inhibitory only at 100-times excess, and histidine or ornithine did not reduce arginine uptake rates significantly. TbAAT16-1 is a high affinity (Km 4.3 ± 0.5 μM) and highly selective L-lysine transporter and of the compounds tested, only L-lysine and thialysine were competing for L-lysine uptake. TbAAT5-3 and TbAAT16-1 are expressed in both procyclic and bloodstream form T. brucei and cMyc-tagged proteins indicate localization at the plasma membrane. RNAi-mediated down-regulation of TbAAT5 and TbAAT16 in bloodstream form trypanosomes resulted in growth arrest, demonstrating that TbAAT5-mediated arginine and TbAAT16-mediated lysine transport are essential for T. brucei. Growth of induced RNAi lines could partially be rescued by supplementing a surplus of arginine or lysine, respectively, while addition of both amino acids was less efficient. Single and double RNAi lines indicate that additional low affinity uptake systems for arginine and lysine are present in T. brucei.
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