Nine neurodegenerative disorders, called polyglutamine (polyQ) diseases, are characterized by the formation of intranuclear amyloid-like aggregates by nine proteins containing a polyQ tract above a threshold length. These insoluble aggregates and/or some of their soluble precursors are thought to play a role in the pathogenesis. The mechanism by which polyQ expansions trigger the aggregation of the relevant proteins remains, however, unclear. In this work, polyQ tracts of different lengths were inserted into a solvent-exposed loop of the β-lactamase BlaP and the effects of these insertions on the properties of BlaP were investigated by a range of biophysical techniques. The insertion of up to 79 glutamines does not modify the structure of BlaP; it does, however, significantly destabilize the enzyme. The extent of destabilization is largely independent of the polyQ length, allowing us to study independently the effects intrinsic to the polyQ length and those related to the structural integrity of BlaP on the aggregating properties of the chimeras. Only chimeras with 55Q and 79Q readily form amyloid-like fibrils; therefore, similarly to the proteins associated with diseases, there is a threshold number of glutamines above which the chimeras aggregate into amyloid-like fibrils. Most importantly, the chimera containing 79Q forms amyloid-like fibrils at the same rate whether BlaP is folded or not, whereas the 55Q chimera aggregates into amyloid-like fibrils only if BlaP is unfolded. The threshold value for amyloid-like fibril formation depends, therefore, on the structural integrity of the β-lactamase moiety and thus on the steric and/or conformational constraints applied to the polyQ tract. These constraints have, however, no significant effect on the propensity of the 79Q tract to trigger fibril formation. These results suggest that the influence of the protein context on the aggregating properties of polyQ disease-associated proteins could be negligible when the latter contain particularly long polyQ tracts.
Haematococcus pluvialis is the current better source of natural astaxanthin, a high-value carotenoid. Traditionally, the production process of astaxanthin by this algae is achieved by a two-stage system: during the first stage, vegetative "green" cells are produced and then converted, in the second stage, into cysts that accumulate astaxanthin. In this work, a medium screening strategy based on the mixing of a 3-component hydroponic fertilizer was applied to identify a new formulation optimized for the vegetative stage. A maximal and high cell density of 2 x 10 6 cells mL −1 was obtained in a medium containing a high level of phosphate relative to nitrate, resulting in a N:P ratio much lower than commonly used media for H. pluvialis. In this medium, cells remained at the vegetative and motile stage during a prolonged period of time. Both high cell density culture and motile stage persistence was proved to be related to the N:P feature of this medium. We conclude that the macrozoid stage of H. pluvialis is favored under high-P and low-N supply and that low-cost hydroponic fertilizers can be successfully used for achieving high density cultures of vegetative cells of H. pluvialis.
Chlorophyll fluorescence is an information-rich signal which provides an access to the management of light absorbed by PSII. A good example of this is the succession of fast fluorescence fluctuations during light-induced photosynthetic induction after dark-adaptation. During this period, the fluorescence trace exhibits several inflexion points: O-J-I-P-S-M-T. Whereas the OJIP part of this kinetics has been the subject of many studies, the processes that underly the PSMT transient are less understood. Here, we report an analysis of the PSMT phase in the green microalga Haematococcus pluvialis in terms of electron acceptors and light use by photochemistry, fluorescence and non-photochemical quenching (NPQ). We identify additional sub-phases between P and S delimited by an inflexion point, that we name Q, found in the second time scale. The P-Q phase expresses a transient photochemical quenching specifically due to alternative electron transport to oxygen. During the transition from Q to S, the NPQ increases and then relaxes during the S-M phase in about 1 min. It is suggested that this transient NPQ observed during induction is a high energy state quenching (qE) dependent on the alternative electron transport to molecular oxygen. We further show that this NPQ is of the same nature than the NPQ, known as the low-wave phenomenon, which is transiently observed after a saturating light pulse given at steady-state. In both cases, the NPQ is oxygen-dependent. This NPQ is observed at external pH 6.0, but not at pH 7.5, which seems correlated with faster saturation of the PQ pool at pH 6.0.
Background The increasing demand for local food production is fueling high interest in the development of controlled environment agriculture. In particular, LED technology brings energy-saving advantages together with the possibility of manipulating plant phenotypes through light quality control. However, optimizing light quality is required for each cultivated plant and specific purpose. Findings This article shows that the combination of LED gradient set-ups with imaging-based non-destructive plant phenotyping constitutes an interesting new screening tool with the potential to improve speed, logistics, and information output. To validate this concept, an experiment was performed to evaluate the effects of a complete range of red:blue ratios on 7 plant species: Arabidopsis thaliana, Brachypodium distachyon, Euphorbia peplus, Ocimum basilicum, Oryza sativa, Solanum lycopersicum, and Setaria viridis. Plants were exposed during 30 days to the light gradient and showed significant, but species-dependent, responses in terms of dimension, shape, and color. A time-series analysis of phenotypic descriptors highlighted growth changes but also transient responses of plant shapes to the red:blue ratio. Conclusion This approach, which generated a large reusable dataset, can be adapted for addressing specific needs in crop production or fundamental questions in photobiology.
Current developments in light-emitting diodes (LEDs) technologies have opened new perspectives for sustainable and highly efficient indoor cultivation. The introduction of LEDs not only allows a reduction in the production costs on a quantitative level, it also offers opportunities to manipulate and optimise qualitative traits. Indeed, while plants respond strongest to red and blue lights for photosynthesis, the whole light spectrum has an effect on plant shape, development, and chemical composition. In order to evaluate LEDs as an alternative to traditional lighting sources, the species-specific plant responses to distinct wavelengths need to be evaluated under controlled conditions. Here, we tested the possibility to use light composition gradients in combination with semi-automated phenotyping to rapidly explore the phenotypic responses of different species to variations in the light spectrum provided by LED sources. Plants of seven different species (Arabidopsis thaliana, Ocimum basilicum, Solanum lycopersicum, Brachypodium distachyon, Oryza sativa, Euphorbia peplus, Setaria viridis) were grown under standard white fluorescent light for 30 days, then transferred to a Red:Blue gradient for another 30 days and finally returned to white light. In all species, differences in terms of dimension, shape, and color were rapidly observed across the gradient and the overall response was widely species-dependent. The experiment yielded large amounts of imaging-based phenotypic data and we suggest simple data analysis methods to aggregate the results and facilitate comparisons between species. Similar experimental setups will help achieve rapid environmental optimization, screen new crop species and genotypes, or develop new gene discovery strategies.
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