Expression of a fungal ferulic acid esterase in suspension cultures of tall fescue (Festuca arundinacea) decreases cell wall feruloylation and increases rates of cell wall digestion

In the cell walls of grasses ferulic acid is esterified to arabinosyl residues in arabinoxylans that can then undergo oxidative coupling reactions to form ferulate dehydrodimers, trimers and oligomers which function to cross-link cell-wall polysaccharides, limiting cell wall degradability. Fungal ferulic acid esterase can release both esterified monomeric and dimeric ferulic acids from these cell wall arabinoxylans making the cell wall more susceptible to further enzymatic attack and increasing cell wall degradability. Non-embryogenic cell suspension cultures of Festuca arundinacea expressing a Aspergillus niger ferulic acid esterase (faeA) targeted to either the apoplast, or endoplasmic reticulum under the control of a constitutive actin promoter, or to the vacuole under the control of a soybean heat shock promoter, were established and FAE activity determined in the cells and medium during a growth cycle. Analysis of the ester-linked ferulates of the cell walls showed that all three transformed cell lines had both reduced ferulate levels and increased levels of xylanase mediated release of wall phenolics on autodigestion as well as increased rates of cell wall digestion in a simulated rumen environment, when compared to control non-transformed cells. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11240-017-1168-9) contains supplementary material, which is available to authorized users

Manipulating the Phenolic Acid Content and Digestibility of Forage Grasses by Targeted Expression of Fungal Cell Wall Degrading Enzymes

Grass cell walls constitute 30-80% of forage dry matter, representing a major source of energy for ruminants. Ferulic acid (4-hydroxy-3-methoxy-cinnamic acid) and other hydroxycinnamic acids are ester linked to arabinosyl residues in arabinoxylans of grass cell walls and undergo oxidative coupling reactions resulting in the formation of a variety of dehydrodiferulate dimers which cross-link cell wall polymers. Although such cross-links have a number of important roles in the cell wall, they also hinder the rate and extent of cell wall degradation by ruminant microbial and fungal enzymes. We have shown previously the expression of a ferulic acid esterase gene from Aspergillus niger in Festuca arundinacea and the potential of the expressed FAE to break phenolic cross-links and release monomeric and dimeric ferulic acids on cell death in vacuole targeted FAE plants. This was enhanced several fold by the addition of exogenous recombinant xylanase (Buanafina et al., 2002). We propose to decrease the level of phenolic cross-linking of cell wall carbohydrate by inducible expression of FAE to the apoplast, ER and golgi and by co-expressing FAE and endo-ß-1,4-xylanase from Trichoderma reesei to the apoplast and vacuole

Projected Rotational Velocities and Stellar Characterization of 350 B Stars in the Nearby Galactic Disk

Projected rotational velocities (vsini) are presented for a sample of 350 early B-type main sequence stars in the nearby Galactic disk. The stars are located within ~1.5 kpc from the Sun, and the great majority within 700 pc. The analysis is based on high-resolution spectra obtained with the MIKE spectrograph on the Magellan Clay 6.5-m telescope at the Las Campanas Observatory in Chile.Spectral types were estimated based on relative intensities of some key line absorption ratios and comparisons to synthetic spectra. Effective temperatures were estimated from the reddening-free Q index, and projected rotational velocities were then determined via interpolation on a published grid that correlates the synthetic full width at half maximum of the He I lines at 4026, 4388 and 4471 A with vsini. As the sample has been selected solely on the basis of spectral types it contains an selection of B stars in the field, in clusters, and in OB associations. The vsini distribution obtained for the entire sample is found to be essentially flat for vsini values between 0-150 km/s, with only a modest peak at low projected rotational velocities. Considering subsamples of stars, there appears to be a gradation in the vsini distribution with the field stars presenting a larger fraction of the slow rotators and the cluster stars distribution showing an excess of stars with vsini between 70 and 130 km/s. Furthermore, for a subsample of potential runaway stars we find that the vsini distribution resembles the distribution seen in denser environments, which could suggest that these runaway stars have been subject to dynamical ejection mechanisms.Comment: 38 pages, 11 figures. Complete sample table. AJ accepte

Retinal glycoprotein enrichment by concanavalin a enabled identification of novel membrane autoantigen synaptotagmin-1 in equine recurrent uveitis.

Complete knowledge of autoantigen spectra is crucial for understanding pathomechanisms of autoimmune diseases like equine recurrent uveitis (ERU), a spontaneous model for human autoimmune uveitis. While several ERU autoantigens were identified previously, no membrane protein was found so far. As there is a great overlap between glycoproteins and membrane proteins, the aim of this study was to test whether pre-enrichment of retinal glycoproteins by ConA affinity is an effective tool to detect autoantigen candidates among membrane proteins. In 1D Western blots, the glycoprotein preparation allowed detection of IgG reactions to low abundant proteins in sera of ERU patients. Synaptotagmin-1, a Ca2+-sensing protein in synaptic vesicles, was identified as autoantigen candidate from the pre-enriched glycoprotein fraction by mass spectrometry and was validated as a highly prevalent autoantigen by enzyme-linked immunosorbent assay. Analysis of Syt1 expression in retinas of ERU cases showed a downregulation in the majority of ERU affected retinas to 24%. Results pointed to a dysregulation of retinal neurotransmitter release in ERU. Identification of synaptotagmin-1, the first cell membrane associated autoantigen in this spontaneous autoimmune disease, demonstrated that examination of tissue fractions can lead to the discovery of previously undetected novel autoantigens. Further experiments will address its role in ERU pathology

Fundamental Parameters of He-Weak and He-Strong Stars

We carried out low resolution spectroscopic observations in the wavelength range 3400-4700 A of 20 He-weak and 8 He-strong stars to determine their fundamental parameters by means of the Divan-Chalonge-Barbier (BCD) spectrophotometric system. For a few He-weak stars we also estimate the effective temperatures and the angular diameters by integrating absolute fluxes observed over a wide spectral range. Non-LTE model calculations are carried out to study the influence of the He/H abundance ratio on the emergent radiation of He-strong stars and on their Teff determination. We find that the effective temperatures, surface gravities and bolometric absolute magnitudes of He-weak stars estimated with the BCD system and the integrated flux method are in good agreement between each other, and they also agree with previous determinations based on several different methods. The mean discrepancy between the visual absolute magnitudes derived using the Hipparcos parallaxes and the BCD values is on average 0.3 mag for He-weak stars, while it is 0.5 mag for He-strong stars. For He-strong stars, we note that the BCD calibration, based on stars in the solar environment, leads to overestimated values of Teff. By means of model atmosphere calculations with enhanced He/H abundance ratios we show that larger He/H ratios produce smaller BD which naturally explains the Teff overestimation. We take advantage of these calculations to introduce a method to estimate the He/H abundance ratio in He-strong stars. The BD of HD 37479 suggests that the Teff of this star remains fairly constant as the star spectrum undergoes changes in the intensity of H and He absorption lines. Data for the He-strong star HD 66765 are reported for the first time.Comment: Accepted for publication in A&

Обзор детекторов гамма-излучения для контроля положения ствола горизонтальной скважины

Представлен обзор детекторов гамма-излучения и принципы их работы. Приведены основные характеристики счетчиков гамма-излучения. Отмечены основные достоинства и недостатки данных устройств. Выбран детектор для регистрации гамма-излучения в процессе горизонтального бурения нефтяных и газовых скважин

Mesoscale features create hotspots of carbon uptake in the Antarctic Circumpolar Current

The influence of eddy structures on the seasonal depletion of dissolved inorganic carbon (DIC) and carbon dioxide (CO2) disequilibrium was investigated during a trans-Atlantic crossing of the Antarctic Circumpolar Current (ACC) in austral summer 2012. The Georgia Basin, downstream of the island of South Georgia (54-55°S, 36-38°W) is a highly dynamic region due to the mesoscale activity associated with the flow of the Subantarctic Front (SAF) and Polar Front (PF). Satellite sea-surface height and chlorophyll-a anomalies revealed a cyclonic cold core that dominated the northern Georgia Basin that was formed from a large meander of the PF. Warmer waters influenced by the SAF formed a smaller anticyclonic structure to the east of the basin. Both the cold core and warm core eddy structures were hotspots of carbon uptake relative to the rest of the ACC section during austral summer. This was most amplified in the cold core where greatest CO2 undersaturation (-78 µatm) and substantial surface ocean DIC deficit (5.1 mol m-2) occurred. In the presence of high wind speeds, the cold core eddy acted as a strong sink for atmospheric CO2 of 25.5 mmol m-2 day-1. Waters of the warm core displayed characteristics of the Polar Frontal Zone (PFZ), with warmer upper ocean waters and enhanced CO2 undersaturation (-59 µatm) and depletion of DIC (4.9mol m-2). A proposed mechanism for the enhanced carbon uptake across both eddy structures is based on the Ekman eddy pumping theory: (i) the cold core is seeded with productive (high chlorophyll-a) waters from the Antarctic Zone and sustained biological productivity through upwelled nutrient supply that counteracts DIC inputs from deep waters; (ii) horizontal entrainment of low-DIC surface waters (biological uptake) from the PFZ downwell within the warm core and cause relative DIC-depletion in the upper water column. The observations suggest that the formation and northward propagation of cold core eddies in the region of the PF could project low-DIC waters towards the site of Antarctic Intermediate Water formation and enhance CO2 drawdown into the deep ocean

Biorefining Potential of Wild-Grown Arundo donax, Cortaderia selloana and Phragmites australis and the Feasibility of White-Rot Fungi-Mediated Pretreatments

Arundo donax, Cortaderia selloana and Phragmites australis are high-biomass-producing perennial Poalean species that grow abundantly and spontaneously in warm temperate regions, such as in Mediterranean-type climates, like those of Southern Europe, Western United States coastal areas, or in regions of South America, South Africa and Australia. Given their vigorous and spontaneous growth, biomass from the studied grasses often accumulates excessively in unmanaged agro-forestry areas. Nonetheless, this also creates the demand and opportunity for the valorisation of these biomass sources, particularly their cell wall polymers, for biorefining applications. By contrast, a related crop, Miscanthus × giganteus, is a perennial grass that has been extensively studied for lignocellulosic biomass production, as it can grow on low-input agricultural systems in colder climates. In this study Fourier transform mid-infrared spectroscopy (FTIR), high-performance anion-exchange chromatography (HPAEC) and lignin content determinations were used for a comparative compositional characterisation of A. donax, C. selloana and P. australis harvested from the wild, in relation to a trial field-grown M. × giganteus high-yielding genotype. A high-throughput saccharification assay showed relatively high sugar release values from the wild-grown grasses, even with a 0.1M NaOH mild alkali pretreatment. In addition to this alkaline pretreatment, biomass was treated with white-rot fungi (WRF), which preferentially degrade lignin more readily than holocellulose. Three fungal species were used: Ganoderma lucidum, Pleurotus ostreatus and Trametes versicolor. Our results showed that neutral sugar contents are not significantly altered, while some lignin is lost during the pretreatments. Furthermore, sugar release upon enzymatic saccharification was enhanced, and this was dependent on the plant biomass and fungal species used in the treatment. To maximise the potential for lignocellulose valorisation, the liquid fractions from the pretreatments were analysed by high performance liquid chromatography – photodiode array detection – electrospray ionisation tandem mass spectrometry (HPLC-PDA-ESI-MS(n)). This study is one of the first to report on the composition of WRF-treated grass biomass, while assessing the potential relevance of breakdown products released during the treatments, beyond more traditional sugar-for-energy applications. Ultimately, we expect that our data will help promote the valorisation of unused biomass resources, create economic value, while contributing to the implementation of sustainable biorefining systems

Measuring the effect of circular public procurement on Government’s Environmental Impact

Industrial Ecolog