Improved understanding of drought controls on seasonal variation in Mediterranean forest canopy CO2 and water fluxes through combined in situ measurements and ecosystem modelling

Water stress is a defining characteristic of Mediterranean ecosystems, and is likely to become more severe in the coming decades. Simulation models are key tools for making predictions, but our current understanding of how soil moisture controls ecosystem functioning is not sufficient to adequately constrain parameterisations. Canopy-scale flux data from four forest ecosystems with Mediterranean-type climates were used in order to analyse the physiological controls on carbon and water flues through the year. Significant non-stomatal limitations on photosynthesis were detected, along with lesser changes in the conductance-assimilation relationship. New model parameterisations were derived and implemented in two contrasting modelling approaches. The effectiveness of two models, one a dynamic global vegetation model ('ORCHIDEE'), and the other a forest growth model particularly developed for Mediterranean simulations ('GOTILWA+'), was assessed and modelled canopy responses to seasonal changes in soil moisture were analysed in comparison with in situ flux measurements. In contrast to commonly held assumptions, we find that changing the ratio of conductance to assimilation under natural, seasonally-developing, soil moisture stress is not sufficient to reproduce forest canopy CO2 and water fluxes. However, accurate predictions of both CO2 and water fluxes under all soil moisture levels encountered in the field are obtained if photosynthetic capacity is assumed to vary with soil moisture. This new parameterisation has important consequences for simulated responses of carbon and water fluxes to seasonal soil moisture stress, and should greatly improve our ability to anticipate future impacts of climate changes on the functioning of ecosystems in Mediterranean-type climates

Large-scale production of somatic embryos as a source of hypocotyl explants for Vitis vinifera micrografting

To the standard methods currently used to make grapevine virus-free, apex micrografting on hypocotyls of somatic embryos is proposed as an alternative procedure. The study defines optimal conditions to produce hypocotyl fragments suitable for micrografting. Interruption of the process by storage of tissues or embryos at low temperature (+ 4 °C) was assessed at different stages and for durations up to 6 months. Best procedure to produce somatic embryos were: long-term maintenance of embryogenic cultures on C1 medium (5 μM 2.4-D + 1 μM BAP, solidified with 4 g·l-1 agar and 4 g·l-1 Phytagel) ; differentiation of embryogenic callus for 2 months on C2 medium (5 μM NOA + 1 μM BAP, gelling agents same as above) ; transfer of single embryos on plant growth regulator-free medium for 2-3 weeks for germination. At different steps of the process, embryogenic tissues or differentiated embryos can be stored for up to 180 d for some cultivars. Micrografting assays were performed with various types of embryo and with apices from several V. vinifera cultivars. White to slightly coloured hypocotyls, excised from embryos germinated in darkness, gave best results for micrografting, while hypocotyl shape had little influence. For all genotypes tested the success rate ranged from 18 to 30 %.

Using bacterial inclusion bodies to screen for amyloid aggregation inhibitors

Background: The amyloid-β peptide (Aβ42) is the main component of the inter-neuronal amyloid plaques characteristic of Alzheimer's disease (AD). The mechanism by which Aβ42 and other amyloid peptides assemble into insoluble neurotoxic deposits is still not completely understood and multiple factors have been reported to trigger their formation. In particular, the presence of endogenous metal ions has been linked to the pathogenesis of AD and other neurodegenerative disorders. Results: Here we describe a rapid and high-throughput screening method to identify molecules able to modulate amyloid aggregation. The approach exploits the inclusion bodies (IBs) formed by Aβ42 when expressed in bacteria. We have shown previously that these aggregates retain amyloid structural and functional properties. In the present work, we demonstrate that their in vitro refolding is selectively sensitive to the presence of aggregation-promoting metal ions, allowing the detection of inhibitors of metal-promoted amyloid aggregation with potential therapeutic interest. Conclusions: Because IBs can be produced at high levels and easily purified, the method overcomes one of the main limitations in screens to detect amyloid modulators: the use of expensive and usually highly insoluble synthetic peptides