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Malinova, Irina ; Mahlow, Sebastian ; Alseekh, Saleh ; Orawetz, Tom ; Fernie, Alisdair ; Baumann, Otto ; Steup, Martin ; Fettke, Jörg
In leaves of two starch-related single-knockout lines lacking either the cytosolic transglucosidase (also designated as disproportionating enzyme 2, DPE2) or the maltose transporter (MEX1), the activity of the plastidial phosphorylase isozyme (PHS1) is increased. In both mutants, metabolism of starch-derived maltose is impaired but inhibition is effective at different subcellular sites. Two constitutive double knockout mutants were generated (designated as dpe2-1 x phs1a and mex1 x phs1b) both lacking functional PHS1. They reveal that in normally grown plants, the plastidial phosphorylase isozyme participates in transitory starch degradation and that the central carbon metabolism is closely integrated into the entire cell biology. All plants were grown either under continuous illumination or in a light-dark regime. Both double mutants were compromised in growth and, compared with the single knockout plants, possess less average leaf starch when grown in a light-dark regime. Starch and chlorophyll contents decline with leaf age. As revealed by transmission electron microscopy, mesophyll cells degrade chloroplasts, but degradation is not observed in plants grown under continuous illumination. The two double mutants possess similar but not identical phenotypes. When grown in a light-dark regime, mesophyll chloroplasts of dpe2-1 x phs1a contain a single starch granule but under continuous illumination more granules per chloroplast are formed. The other double mutant synthesizes more granules under either growth condition. In continuous light, growth of both double mutants is similar to that of the parental single knockout lines. Metabolite profiles and oligoglucan patterns differ largely in the two double mutants.
Balazadeh, Salma ; Schildhauer, Joerg ; Araujo, Wagner L. ; Munne-Bosch, Sergi ; Fernie, Alisdair ; Proost, Sebastian ; Humbeck, Klaus ; Müller-Röber, Bernd
Leaf senescence is a developmentally controlled process, which is additionally modulated by a number of adverse environmental conditions. Nitrogen shortage is a well-known trigger of precocious senescence in many plant species including crops, generally limiting biomass and seed yield. However, leaf senescence induced by nitrogen starvation may be reversed when nitrogen is resupplied at the onset of senescence. Here, the transcriptomic, hormonal, and global metabolic rearrangements occurring during nitrogen resupply-induced reversal of senescence in Arabidopsis thaliana were analysed. The changes induced by senescence were essentially in keeping with those previously described; however, these could, by and large, be reversed. The data thus indicate that plants undergoing senescence retain the capacity to sense and respond to the availability of nitrogen nutrition. The combined data are discussed in the context of the reversibility of the senescence programme and the evolutionary benefit afforded thereby. Future prospects for understanding and manipulating this process in both Arabidopsis and crop plants are postulated.
Pieper, Imke ; Wehe, Christoph A. ; Bornhorst, Julia ; Ebert, Franziska ; Leffers, Larissa ; Holtkamp, Michael ; Höseler, Pia ; Weber, Till ; Mangerich, Aswin ; Bürkle, Alexander ; Karst, Uwe ; Schwerdtle, Tanja
The toxicologically most relevant mercury (Hg) species for human exposure is methylmercury (MeHg). Thiomersal is a common preservative used in some vaccine formulations. The aim of this study is to get further mechanistic insight into the yet not fully understood neurotoxic modes of action of organic Hg species. Mercury species investigated include MeHgCl and thiomersal. Additionally HgCl2 was studied, since in the brain mercuric Hg can be formed by dealkylation of the organic species. As a cellular system astrocytes were used. In vivo astrocytes provide the environment necessary for neuronal function. In the present study, cytotoxic effects of the respective mercuricals increased with rising alkylation level and correlated with their cellular bioavailability. Further experiments revealed for all species at subcytotoxic concentrations no induction of DNA strand breaks, whereas all species massively increased H2O2-induced DNA strand breaks. This co-genotoxic effect is likely due to a disturbance of the cellular DNA damage response. Thus, at nanomolar, sub-cytotoxic concentrations, all three mercury species strongly disturbed poly(ADP-ribosyl)ation, a signalling reaction induced by DNA strand breaks. Interestingly, the molecular mechanism behind this inhibition seems to be different for the species. Since chronic PARP-1 inhibition is also discussed to sacrifice neurogenesis and learning abilities, further experiments on neurons and in vivo studies could be helpful to clarify whether the inhibition of poly(ADP-ribosyl)ation contributes to organic Hg induced neurotoxicity.
Pieper, Imke ; Wehe, Christoph A. ; Bornhorst, Julia ; Ebert, Franziska ; Leffers, Larissa ; Holtkamp, Michael ; Höseler, Pia ; Weber, Till ; Mangerich, Aswin ; Bürkle, Alexander ; Karst, Uwe ; Schwerdtle, Tanja
The toxicologically most relevant mercury (Hg) species for human exposure is methylmercury (MeHg). Thiomersal is a common preservative used in some vaccine formulations. The aim of this study is to get further mechanistic insight into the yet not fully understood neurotoxic modes of action of organic Hg species. Mercury species investigated include MeHgCl and thiomersal. Additionally HgCl2 was studied, since in the brain mercuric Hg can be formed by dealkylation of the organic species. As a cellular system astrocytes were used. In vivo astrocytes provide the environment necessary for neuronal function. In the present study, cytotoxic effects of the respective mercuricals increased with rising alkylation level and correlated with their cellular bioavailability. Further experiments revealed for all species at subcytotoxic concentrations no induction of DNA strand breaks, whereas all species massively increased H2O2-induced DNA strand breaks. This co- genotoxic effect is likely due to a disturbance of the cellular DNA damage response. Thus, at nanomolar, sub-cytotoxic concentrations, all three mercury species strongly disturbed poly(ADP-ribosyl)ation, a signalling reaction induced by DNA strand breaks. Interestingly, the molecular mechanism behind this inhibition seems to be different for the species. Since chronic PARP-1 inhibition is also discussed to sacrifice neurogenesis and learning abilities, further experiments on neurons and in vivo studies could be helpful to clarify whether the inhibition of poly(ADP-ribosyl) ation contributes to organic Hg induced neurotoxicity.
Leaching of dissolved C in arable hummocky ground moraine soil landscapes is characterized by a spatial continuum of more or less erosion-affected Luvisols, Calcaric Regosols at exposed positions, and Colluvic Regosols in depressions. Our objective was to estimate the fluxes of dissolved C in four differently eroded soils as affected by erosion-induced pedological and soil structural alterations. In this model study, we considered landscape position effects by adapting the water table as the bottom boundary condition and erosion effects by using pedon-specific soil hydraulic properties. The one-dimensional vertical water movement was described with the Richards equation using HYDRUS-1D. Solute fluxes were obtained by combining calculated water fluxes with concentrations of dissolved organic and inorganic C (DOC and DIC, respectively) measured from soil solution extracted by suction cups at biweekly intervals. In the 3-yr period (2010-2012), DOC fluxes in the 2-m soil depth were similar at the three non-colluvic locations with -0.8 +/- 0.1 g m(-2) yr(-1) (i.e., outflow) but were 0.4 g m(-2) yr(-1) (i.e., input) in the depression. The DIC fluxes ranged from -10.2 g m(-2) yr(-1) for the eroded Luvisol, -9.2 g m(-2) yr(-1) for the Luvisol, and -6.1 g m(-2) yr(-1) for the Calcaric Regosol to 3.2 g m(-2) yr(-1) for the Colluvic Regosol. The temporal variations in DOC and DIC fluxes were controlled by water fluxes. The spatially distributed leaching results corroborate the hypothesis that the effects of soil erosion influence fluxes through modified hydraulic and transport properties and terrain-dependent boundary conditions.
The size and dynamics of biogenic silicon (BSi) pools influence silicon (Si) fluxes from terrestrial to aquatic ecosystems. The research focus up to now was on the role of plants in Si cycling. In recent studies on old forests annual biosilicification rates of idiosomic testate amoebae (i.e. TA producing self-secreted silica shells) were shown to be of the order of Si uptake by trees. However, no comparable data exist for initial ecosystems. We analyzed the protozoic BSi pool (idiosomic TA), corresponding annual biosilicification rates and readily available and amorphous Si fractions along a 10-year chronosequence in a post-mining landscape in Brandenburg, Germany.
Idiosomic Si pools ranged from 3 to 680 g Si ha(-1) and were about 3-4 times higher at vegetated compared to uncovered spots. They increased significantly with age and were related to temporal development of soil chemical properties. The calculation of annual biosilicification resulted in maxima between 2 and 16 kg Si ha(-1) with rates always higher at vegetated spots. Our results showed that the BSi pool of idiosomic TA is built up rapidly during the initial phases of ecosystem development and is strongly linked to plant growth. Furthermore, our findings highlight the importance of TA for Si cycling in young artificial ecosystems. (C) 2014 Elsevier B.V. All rights reserved.