Isolation of a new Papiliotrema laurentii strain that displays capacity to achieve high lipid content from xylose.

In this work, we isolated and selected oleaginous yeasts from rock field soils from two National Parks in Brazil (Caparaó and Serra dos Órgãos) with the potential to accumulate oil from xylose, the main pentose sugar found in lignocellulosic biomass. From the 126 isolates, two were selected based on their lipid contents. They were taxonomically identified as Papiliotrema laurentii (UFV-1 and UFV-2). Of the two, P. laurentii UFV-1 was selected as the best lipid producer. Under unoptimized conditions, lipid production by P. laurentii UFV-1 was higher in glucose than in xylose. To improve its lipid production from xylose, we applied response surface methodology (RSM) with a face-centered central composite design (CCF). We evaluated the effects of agitation rate, initial cell biomass (OD600), carbon/nitrogen ratio (C/N ratio) and pH on lipid production. P. laurentii UFV-1 recorded the highest lipid content, 63.5% (w/w) of the cell dry mass, under the following conditions: C/N ratio = 100:1, pH value = 7.0, initial OD600 = 0.8 and agitation = 300 rpm. Under these optimized conditions, biomass, lipid titer and volumetric lipid productivity were 9.31 g/L, 5.90 g/L and 0.082 g/L.h, respectively. Additionally, we determined the fatty acid composition of P. laurentii UFV-1 as follows: C14:0 (0.5%), C16:0 (28.4-29.4%), C16:1 (0.2%), C18:0 (9.5-11%), C18:1 (58.6-60.5%), and C20:0 (0.7-0.8%). Based on this composition, the predicted properties of biodiesel showed that P. laurentii UFV-1 oil is suitable for use as feedstock in biodiesel production.

How do wheat plants cope with Pyricularia oryzae infection? A physiological and metabolic approach.

MAIN CONCLUSION: The infection of wheat leaves by Pyricularia oryzae induced remarkable reprogramming of the primary metabolism (amino acids, sugars, and organic acids) in favor of a successful fungal infection and certain changes were conserved among cultivars regardless of their level of resistance to blast. Wheat blast, caused by Pyricularia oryzae, has become one of the major threats for food security worldwide. Here, we investigated the behavior of three wheat cultivars (BR-18, Embrapa-16, and BRS-Guamirim), differing in their level of resistance to blast, by analyzing changes in cellular damage, antioxidative metabolism, and defense compounds as well as their photosynthetic performance and metabolite profile. Blast severity was lower by 45 and 33% in Embrapa-16 and BR-18 cultivars (moderately resistant), respectively, at 120 h after inoculation in comparison to BRS-Guamirim cultivar (susceptible). Cellular damage caused by P. oryzae infection was great in BRS-Guamirim compared to BR-18. The photosynthetic performance of infected plants was altered due to diffusional and biochemical limitations for CO2 fixation. At the beginning of the infection process, dramatic changes in both carbohydrate metabolism and on the levels of amino acids, intermediate compounds of the tricarboxylic acid cycle, and polyamines were noticed regardless of cultivar suggesting an extensive metabolic reprogramming of the plants following fungal infection. Nevertheless, Embrapa-16 plants displayed a more robust and efficient antioxidant metabolism, higher phenylalanine ammonia-lyase and polyphenoloxidase activities and higher concentrations of phenolics and lignin, which, altogether, helped them to counteract more efficiently the infection by P. oryzae. Our results demonstrated that P. oryzae infection significantly modified the metabolism of wheat plants and different types of metabolic defence may act both additively and synergistically to provide additional plant protection to blast.

Ethanol stress responses of Kluyveromyces marxianus CCT 7735 revealed by proteomic and metabolomic analyses.

Kluyveromyces marxianus CCT 7735 offers advantages to ethanol production over Saccharomyces cerevisiae, including thermotolerance and the ability to convert lactose to ethanol. However, its growth is impaired at high ethanol concentrations. Herein we report on the protein and intracellular metabolite profiles of K. marxianus at 1 and 4 h under ethanol exposure. The concentration of some amino acids, trehalose and ergosterol were also measured. We observed that proteins and metabolites from carbon pathways and translation were less abundant, mainly at 4 h of ethanol stress. Nevertheless, the concentration of some amino acids and trehalose increased at 8 and 12 h under ethanol stress, indicating an adaptive response. Moreover, our results show that the abundance of proteins and metabolites related to the oxidative stresses responses increased. The results obtained in this study provide insights into understanding the physiological changes in K. marxianus under ethanol stress, indicating possible targets for ethanol tolerant strains construction.

Validation and application of a QuEChERS based method for estimation of half-life of imidazolinone herbicides in soils by LC-ESI-MS/MS.

A modified QuEChERS (quick, easy, cheap, effective, rugged and safe) method was validated and applied for the estimation of half-lives of two imidazolinone group herbicides, imazethapyr and imazapic, in the Dystric Plinthosol (FX) and Ferralsol (LVA) agricultural soils using liquid chromatography tandem mass spectrometry. The solutions were prepared in a matrix extract to avoid matrix effects. The analytical method showed satisfactory specificity, selectivity, linearity (R2 > 0.99), recoveries (range 85.0-117.0%), with RSD between 6.7% and 16.6%, and precision (range 94.7-108.5%), with RSD between 6.0% and 11.9%. The limit of detections for imazethapyr and imazapic in the soils were 2.2 µg kg-1 and 2.0 µg kg-1, and the limit of quantifications were 6.6 µg kg-1 and 6.1 µg kg-1. The half-lives of imazethapyr (35.7 and 97.9 days) and imazapic (40.4 and 64.4 days), in the FX and LVA soils, respectively, indicate that they are medium-persistence herbicides with possibility of leaching into groundwater. In addition, high concentrations of imazethapyr and imazapic were found in the soil samples after the time of application, meaning that there is a potential for prolonged soil residual activity due to carryover.

Transcriptome analysis of the thermotolerant yeast Kluyveromyces marxianus CCT 7735 under ethanol stress.

The thermotolerant yeast Kluyveromyces marxianus displays a potential to be used for ethanol production from both whey and lignocellulosic biomass at elevated temperatures, which is highly alluring to reduce the cost of the bioprocess. Nevertheless, contrary to Saccharomyces cerevisiae, K. marxianus cannot tolerate high ethanol concentrations. We report the transcriptional profile alterations in K. marxianus under ethanol stress in order to gain insights about mechanisms involved with ethanol response. Time-dependent changes have been characterized under the exposure of 6% ethanol and compared with the unstressed cells prior to the ethanol addition. Our results reveal that the metabolic flow through the central metabolic pathways is impaired under the applied ethanol stress. Consistent with these results, we also observe that genes involved with ribosome biogenesis are downregulated and gene-encoding heat shock proteins are upregulated. Remarkably, the expression of some gene-encoding enzymes related to unsaturated fatty acid and ergosterol biosynthesis decreases upon ethanol exposure, and free fatty acid and ergosterol measurements demonstrate that their content in K. marxianus does not change under this stress. These results are in contrast to the increase previously reported with S. cerevisiae subjected to ethanol stress and suggest that the restructuration of K. marxianus membrane composition differs in the two yeasts which gives important clues to understand the low ethanol tolerance of K. marxianus compared to S. cerevisiae.