Project description:Plant material consisted of synthetic hexaploid wheat germplasm into the Opata background (Altar 84/ Aegilops squarrosa (TAUS)//Opata) . Plants were grown at a density of 9-11 individuals per 20cm x 10cm (diameter x height) plastic pot containing 1500g well-rinsed Turface MVP® medium (Profile Products LLC, Buffalo, IL), in controlled environment chambers at 23°C, 70% relative humidity, and 16h photoperiod with a photosynthetic photon flux (PPF) of 330±10 µmolem-2s-1 when measured at the top of the canopy at growth stage 22 to 24 in the Zadoks scale (Zadoks et al., 1974). Plants were watered daily until 21 days after seeding (DAS) by flooding trays with water for 5 minutes, then draining the trays. Drought stress was applied by water withholding, beginning on 21 DAS. Watering was withheld from plants belonging to drought treatment, while the control group received water above field capacity. Water content of the growth medium was gravimetrically monitored. Root tissues were collected from control and treated plants when the water content of the treatment group growth medium fell below the permanent wilting point. Three biological replicates were conducted in three separate time-span courses. Total RNA was isolated using a LiCl3 precipitation method following Moore et al., (2005). Dye swap desin microarray analyses of well-watered and water-stressed root tissues were conducted across three biological replicates totaling six hybridizations.
Project description:Water kefir is a slightly alcoholic and traditionally fermented beverage, which is prepared from sucrose, water, kefir grains, and dried or fresh fruits (e.g. figs). Lactobacillus (L.) nagelii, L. hordei and Saccharomyces (S.) cerevisiae are predominant and stable lactic acid bacteria and yeasts, respectively, isolated from water kefir consortia. The growth of L. nagelii and L. hordei are improved in the presence of S. cerevisiae. In this work we demonstrate that quantitative comparative proteomics enables the investigation of interactions between LAB and yeast to predict real-time metabolic exchange in water kefir. It revealed 73 differentially expressed (DE) in L. nagelii TMW 1.1827 in the presence of S. cerevisiae. The presence of the yeast induced changes in the changes in the carbohydrate metabolism of L. nagelii and affected reactions involved in NAD+/NADH homeostasis. Furthermore, the DE enzymes involved in amino acid biosynthesis or catabolism predict that S. cerevisiae releases glutamine, histidine, methionine and arginine, which are subsequently used by L. nagelii to ensure its survival in the water kefir consortium. In co-culture with S. cerevisiae, L. nagelii profits from riboflavin, most likely secreted by the yeast. The reaction of L. nagelii to the presence of S. cerevisiae differs from that one of the previously studied L. hordei, which displays 233 differentially expressed proteins, changes in citrate metabolism and an antidromic strategy for NAD+/NADH homeostasis. So far, aggregation promotion factors, i.e. formation of a specific glucan and bifunctional enzymes were only detected in L. hordei.
Project description:<p><strong>INTRODUCTION:</strong> The extraction solvent mixtures were optimized for untargeted metabolomics analysis of microbial communities from two laboratory scale activated sludge reactors performing enhanced biological phosphorus removal (EBPR).</p><p><strong>OBJECTIVE:</strong> To develop a robust and simple analytical protocol to analyse microbial metabolomics from EBPR bioreactors.</p><p><strong>METHODS:</strong> Extra- and intra-cellular metabolites were extracted using five methods and analysed by ultraperformance liquid chromatography mass spectrometry (UPLC-MS).</p><p><strong>RESULTS:</strong> The optimal extraction method was biomass specific and methanol:water (1:1 v/v) and methanol:chloroform:water (2:2:1 v/v) were chosen, respectively, for each of the two different bioreactors.</p><p><strong>CONCLUSION:</strong> Our approach provides direct surveys of the metabolic state of PAO-enriched EBPR communities, showing that extraction methods should be carefully tailored to the microbial community under study</p>