Thermoresponsive structures in the 5-untranslated region of mRNA are known to control translation of heat shock and virulence genes. Expression of many rhizobial heat shock genes is regulated by a conserved sequence element called ROSE for repression of heat shock gene expression. This cis-acting, untranslated mRNA is thought to prevent ribosome access at low temperature through an extended secondary structure, which partially melts when the temperature rises. We show here by a series of in vivo and in vitro approaches that ROSE is a sensitive thermometer responding in the physiologically relevant temperature range between 30 and 40°C. Point mutations predicted to disrupt base pairing enhanced expression at 30°C. Compensatory mutations restored repression, emphasizing the importance of secondary structures in the sensory RNA. Only moderate inducibility of a 5-truncated ROSE variant suggests that interactions between individual stem loops coordinate temperature sensing. In the presence of a complementary oligonucleotide, the functionally important stem loop of ROSE was rendered susceptible to RNase H treatment at heat shock temperatures. Since major structural rearrangements were not observed during UV and CD spectroscopy, subtle structural changes involving the Shine-Dalgarno sequence are proposed to mediate translational control. Temperature perception by the sensory RNA is an ordered process that most likely occurs without the aid of accessory factors.

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International audienceBiomass can be converted into liquid and gaseous biofuels with good efficiency. In this study, the conversion of industrial hemp ( L.), a biomass source that can be cultivated with a high biomass yield per hectare, was used. Steam pretreatment of dry and ensiled hemp was investigated prior to ethanol production. The pretreatment efficiency was evaluated in terms of sugar recovery and polysaccharide conversion in the enzymatic hydrolysis step. For both materials, impregnation with 2% SO followed by steam pretreatment at 210°C for five minutes were found to be the optimal conditions leading to the highest overall yield of glucose. Simultaneous saccharification and fermentation experiments carried out with optimised pretreatment conditions resulted in ethanol yields of 163 g kg ensiled hemp (dry matter) (71% of the theoretical maximum) and 171 g kg dry hemp (74%), which corresponds to 206-216 l Mg ethanol based on initial dry material

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BackgroundEnsiling is a common method of preserving energy crops for anaerobic digestion, and many scientific studies report that ensiling increases the methane yield. In this study, the ensiling process and the methane yields before and after ensiling were studied for four crop materials.ResultsThe changes in wet weight and total solids (TS) during ensiling were small and the loss of energy negligible. The methane yields related to wet weight and to volatile solids (VS) were not significantly different before and after ensiling when the VS were corrected for loss of volatile compounds during TS and VS determination. However, when the TS were measured according to standard methods and not corrected for losses of volatile compounds, the TS loss during ensiling was overestimated for maize and sugar beet. The same methodological error leads to overestimation of methane yields; when TS and VS were not corrected the methane yield appeared to be 51% higher for ensiled than fresh sugar beet.ConclusionsEnsiling did not increase the methane yield of the studied crops. Published methane yields, as well as other information on silage related to uncorrected amounts of TS and VS, should be regarded with caution.

BackgroundCurrent EU directives demand increased use of renewable fuels in the transportation sector but restrict governmental support for production of biofuels produced from crops. The use of intercropped lucerne and wheat may comply with the directives. In the current study, the combination of ensiled lucerne (Medicago sativa L.) and wheat straw as substrate for hydrogen and methane production was investigated. Steam-pretreated and enzymatically hydrolysed wheat straw [WSH, 76% of total chemical oxygen demand (COD)] and ensiled lucerne (LH, 24% of total COD) were used for sequential hydrogen production through dark fermentation and methane production through anaerobic digestion and directly for anaerobic digestion. Synthetic co-cultures of extreme thermophilic Caldicellulosiruptor species adapted to elevated osmolalities were used for dark fermentation.ResultsBased on 6 tested steam pretreatment conditions, 5 min at 200 °C was chosen for the ensiled lucerne. The same conditions as applied for wheat straw (10 min at 200 °C with 1% acetic acid) would give similar sugar yields. Volumetric hydrogen productivities of 6.7 and 4.3 mmol/L/h and hydrogen yields of 1.9 and 1.8 mol/mol hexose were observed using WSH and the combination of WSH and LH, respectively, which were relatively low compared to those of the wild-type strains. The combinations of WSH plus LH and the effluent from dark fermentation of WSH plus LH were efficiently converted to methane in anaerobic digestion with COD removal of 85–89% at organic loading rates of COD 5.4 and 8.5 g/L/day, respectively, in UASB reactors. The nutrients in the combined hydrolysates could support this conversion.ConclusionsThis study demonstrates the possibility of reducing the water addition to WSH by 26% and the phosphorus addition by 80% in dark fermentation with Caldicellulosiruptor species, compared to previous reports. WSH and combined WSH and LH were well tolerated by osmotolerant co-cultures. The yield was not significantly different when using defined media or hydrolysates with the same concentrations of sugars. However, the sugar concentration was negatively correlated with the hydrogen yield when comparing the results to previous reports. Hydrolysates and effluents from dark fermentation can be efficiently converted to methane. Lucerne can serve as macronutrient provider in anaerobic digestion. Intercropping with wheat is promising.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1280-z) contains supplementary material, which is available to authorized users.

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