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Rüdiger und die Gaben
(2019)
Identification and Characterization of Three Epithiospecifier Protein Isoforms in Brassica oleracea
(2019)
Glucosinolates present in Brassicaceae play a major role in herbivory defense. Upon tissue disruption, glucosinolates come into contact with myrosinase, which initiates their breakdown to biologically active compounds. Among these, the formation of epithionitriles is triggered by the presence of epithiospecifier protein (ESP) and a terminal double bond in the glucosinolate side chain. One ESP gene is characterized in the model plant Arabidopsis thaliana (AtESP; At1g54040.2). However, Brassica species underwent genome triplication since their divergence from the Arabidopsis lineage. This indicates the presence of multiple ESP isoforms in Brassica crops that are currently poorly characterized. We identified three B. oleracea ESPs, specifically BoESP1 (LOC106296341), BoESP2 (LOC106306810), and BoESP3 (LOC106325105) based on in silico genome analysis. Transcript and protein abundance were assessed in shoots and roots of four B. oleracea vegetables, namely broccoli, kohlrabi, white, and red cabbage, because these genotypes showed a differential pattern for the formation of glucosinolate hydrolysis products as well for their ESP activity. BoESP1 and BoESP2 were expressed mainly in shoots, while BoESP3 was abundant in roots. Biochemical characterization of heterologous expressed BoESP isoforms revealed different substrate specificities towards seven glucosinolates: all isoforms showed epithiospecifier activity on alkenyl glucosinolates, but not on non-alkenyl glucosinolates. The pH-value differently affected BoESP activity: while BoESP1 and BoESP2 activities were optimal at pH 6-7, BoESP3 activity remained relatively stable from pH 4 to 7. In order test their potential for the in vivo modification of glucosinolate breakdown, the three isoforms were expressed in A. thaliana Hi-0, which lacks AtESP expression, and analyzed for the effect on their respective hydrolysis products. The BoESPs altered the hydrolysis of allyl glucosinolate in the A. thaliana transformants to release 1-cyano-2,3-epithiopropane and reduced formation of the corresponding 3-butenenitrile and allyl isothiocyanate. Plants expressing BoESP2 showed the highest percentage of released epithionitriles. Given these results, we propose a model for isoform-specific roles of B. oleracea ESPs in glucosinolate breakdown.
Large-magnitude fluid-injection induced seismic events are a potential risk for geothermal energy developments worldwide. One potential risk mitigation measure is the application of cyclic injection schemes. After validation at small (laboratory) and meso (mine) scale, the concept has now been applied for the first time at field scale at the Pohang Enhanced Geothermal System (EGS) site in Korea. From 7 August until 14 August 2017 a total of 1756 m(3) of surface water was injected into Pohang well PX-1 at flow rates between 1 and 10 l s(-1), with a maximum wellhead pressure (WHP) of 22.8 MPa, according to a site-specific cyclic soft stimulation schedule and traffic light system. A total of 52 induced microearthquakes were detected in real-time during and shortly after the injection, the largest of M-w 1.9. After that event a total of 1771 m(3) of water was produced back from the well over roughly 1 month, during which time no larger-magnitude seismic event was observed. The hydraulic data set exhibits pressure-dependent injectivity increase with fracture opening between 15 and 17 MPa WHP, but no significant permanent transmissivity increase was observed. The maximum magnitude of the induced seismicity during the stimulation period was below the target threshold of M-w 2.0 and additional knowledge about the stimulated reservoir was gained. Additionally, the technical feasibility of cyclic injection at field scale was evaluated. The major factors that limited the maximum earthquake magnitude are believed to be: limiting the injected net fluid volume, flowback after the occurrence of the largest induced seismic event, using a cyclic injection scheme, the application of a traffic light system, and including a priori information from previous investigations and operations in the treatment design.
Saturn’s main ring system is associated with a set of small moons that either are embedded within it or interact with the rings to alter their shape and composition. Five close flybys of the moons Pan, Daphnis, Atlas, Pandora, and Epimetheus were performed between December 2016 and April 2017 during the ring-grazing orbits of the Cassini mission. Data on the moons’ morphology, structure, particle environment, and composition were returned, along with images in the ultraviolet and thermal infrared. We find that the optical properties of the moons’ surfaces are determined by two competing processes: contamination by a red material formed in Saturn’s main ring system and accretion of bright icy particles or water vapor from volcanic plumes originating on the moon Enceladus.