Identification of a UDP-glucosyltransferase conferring deoxynivalenol resistance in Aegilops tauschii and wheat

Aegilops tauschii is the diploid progenitor of the wheat D subgenome and a valuable resource for wheat breeding, yet, genetic analysis of resistance against Fusarium head blight (FHB) and the major Fusarium mycotoxin deoxynivalenol (DON) is lacking. We treated a panel of 147 Ae. tauschii accessions with either Fusarium graminearum spores or DON solution and recorded the associated disease spread or toxin-induced bleaching. A k-mer-based association mapping pipeline dissected the genetic basis of resistance and identified candidate genes. After DON infiltration nine accessions revealed severe bleaching symptoms concomitant with lower conversion rates of DON into the non-toxic DON-3-O-glucoside. We identified the gene AET5Gv20385300 on chromosome 5D encoding a uridine diphosphate (UDP)-glucosyltransferase (UGT) as the causal variant and the mutant allele resulting in a truncated protein was only found in the nine susceptible accessions. This UGT is also polymorphic in hexaploid wheat and when expressed in Saccharomyces cerevisiae only the full-length gene conferred resistance against DON. Analysing the D subgenome helped to elucidate the genetic control of FHB resistance and identified a UGT involved in DON detoxification in Ae. tauschii and hexaploid wheat. This resistance mechanism is highly conserved since the UGT is orthologous to the barley UGT HvUGT13248 indicating descent from a common ancestor of wheat and barley.     

Über die Einwirkung Hypotonischer Medien auf das Zwischenhirn-Hypophysensystem von Mugil und Gobius

Zusammenfassung Bei den Teleostiern Mugil und Gobius wurden die Veränderungen des Zwischenhirn-Hypophysensystems in veränderten Außenmedien (hypotonisches Seewasser) histologisch untersucht.Beim langsamen Überführen aus Seewasser von 38,4 S in Seewasser von 25,4 und 20,8 S nahm das Sekret nicht, wie erwartet, zu. Im Nucleus praeopticus und im Hinterlappen der Hypophyse erfolgt eine starke Verringerung des Sekretbestandes (Mugil). Erst in Seewasser von weit niedrigerem Salzgehalt (6,3 und 1, 4) nimmt das Sekret im Hypophysenhinterlappen wieder zu (Gobius). Bei 6 S ist eine intensive Sekretableitung im N.p. und Tractus praeopticohypophyseus zu beobachten. Eine Deutung der Ergebnisse wird versucht, indem das weitgehend konstante Innenmedium mit den wechselnden Außenfaktoren in Beziehung gebracht wird.Die Befunde weisen darauf hin, daß das Zwischenhirn-Hypophysensystem auch bei Teleostiern an der Regulation des Wasserhaushaltes beteiligt ist.Der Deutschen Forschungsgemeinschaft danke ich für die Gewährung eines Stipendiums, das mir den Aufenthalt in Neapel ermöglichte. Mein besonderer Dank gilt auch den Herren Prof. Dr. Remane, Prof. Dr. Bargmann und Dozent Dr. Schiebler für ihre Unterstützung.     

Untersuchungenüber Das tageszeitlich gebundene Schlüpfen yon Pseudosmittia Arenaria (Dipt. Chironomidae)

Zusammenfassung In einleitenden terminologischen Erwägungen wird vorgeschlagen, den Ausdruck Schlüpfrhythmus zugunsten von tageszeitlich gebundenem Schlüpfen aufzugeben. Pseudosmittia arenaria zeigt im normalen Tag-Nachtwechsel ein deutliches tageszeitlich gebundenes Schlüpfen mit einem Maximum 6–8h nach DL (Beginn der Beleuchtung).In Dauerdunkel schlüpfen keine Imagines. In Dauerlicht erscheinen die Tiere gleichmäßig über den ganzen Tag verteilt.Der ganze Bereich des sichtbaren Lichtes (geprüft von 476–641 m) ist wirksam. Auch die Lichtintensität in Licht-Dunkelbedingungen spielt im untersuchten Bereich (18–350 Lux) keine Rolle.Das Schlüpfmaximum von Ps. arenaria zeigt eine deutliche Beziehung zu DL. Es wandert bei konstanter Tageslänge um so dichter an DL heran, je kürzer die relative Länge der Lichtzeit ist.Bei kürzeren Tageslängen wandert das Maximum von DL fort, bei längeren an DL heran. Bei Tagen, die kürzer als 18h sind, liegt es in der folgenden Dunkelzeit, bei solchen, die länger als 36h sind, vor DL. Sein Weg beschreibt dabei eine kubische Parabel.Beim Eintritt des Gipfels in die folgende Dunkelzeit erscheint ein Maximum nur an jedem 2. Tag. Ein Maximum vor DL ruft ein zusätzliches Maximum in der gleichen Entfernung von LD (Beginn der Dunkelzeit) hervor. Bei extremen Tageslängen ist also dennoch ein Maximum ungefähr alle 24h zu beobachten. Dies ist exogen bedingt. Verlagert man durch schwache Beleuchtung während der Dunkelzeit das Maximum im 12h-Tag in die Lichtzeit, so erhält man ein Maximum an jedem Tag. Der Eintritt des Maximums in eine Dunkelzeit hat also das Umspringen auf einen Schlüpfgipfel an jedem 2. Tag bzw. auf 2 je Tag zur Folge.Im 12h-Tag mit absoluter Dunkelzeit während der Dunkelzeit kann man auch ein Maximum alle 12h erreichen, und zwar durch Umstimmung eines Teils der Population. Man hat dann 2 Gruppen, deren Maxima um 12h gegeneinander verschoben sind. Hemmend und fördernd auf den Einfluß des Lichtes wirken Temperatur und Substratfeuchtigkeit. Beide Faktoren können ohne Licht kein Schlüpfen hervorrufen. Beziehungen zwischen Lebensweise, Ökologie oder systematischer Stellung und tageszeitlich gebundenem Schlüpfen lassen sich bisher nicht feststellen.     

Effects of oxygen level on metabolism and development of seedlings of Trapa natans and two ecologically related species

Seeds of the water plant Trapa natans L. (water chestnut) can germinate in strict anoxia. The seedlings show seminal roots growing upwards while shoot buds remain quiescent until O₂ becomes available. Trapa seedlings are highly tolerant to anoxia. The rate of ethanol fermentation was 21.2 μmol (g FW)⁻¹ h⁻¹, while production of lactate was negligible and lower than that of succinate. The seminal root of Trapa compares better to the rice coleoptile rather than to the rice root, both functionally and as to the metabolic response to anoxia. The anaerobic germination of Nuphar luteum L. and Scirpus mucronatus L. was also characterized by a limited developmental program.     

First report on new evidence for the occurrence of Podocarpus and possible human presence at the mouth of the Amazon during the Late-glacial

Palynological studies on late Quaternary lake sediments from the region of the Amazon estuary, 100 km north-east of Belém, Pará State, Brazil, enable reconstruction of lowland Amazonian rain forest during the Late-glacial and Holocene periods. Late-glacial forests included populations of Podocarpus which suggests a distinct climatic cooling. Ilex was abundant in the early Holocene. Records of the mangrove taxon, Rhizophora, indicate rapid Atlantic sea-level rise in the beginning of the Holocene. High charcoal representation may reflect the first arrival of Amerindians in the Amazon coastal area, probably about 10 800 B.P.     

snRNP Sm proteins share two evolutionarily conserved sequence motifs which are involved in Sm protein-protein interactions.

The spliceosomal small nuclear ribonucleoproteins (snRNPs) U1, U2, U4/U6 and U5 share eight proteins B', B, D1, D2, D3, E, F and G which form the structural core of the snRNPs. This class of common proteins plays an essential role in the biogenesis of the snRNPs. In addition, these proteins represent the major targets for the so-called anti-Sm auto-antibodies which are diagnostic for systemic lupus erythematosus (SLE). We have characterized the proteins F and G from HeLa cells by cDNA cloning, and, thus, all human Sm protein sequences are now available for comparison. Similar to the D, B/B' and E proteins, the F and G proteins do not possess any of the known RNA binding motifs, suggesting that other types of RNA-protein interactions occur in the snRNP core. Strikingly, the eight human Sm proteins possess mutual homology in two regions, 32 and 14 amino acids long, that we term Sm motifs 1 and 2. The Sm motifs are evolutionarily highly conserved in all of the putative homologues of the human Sm proteins identified in the data base. These results suggest that the Sm proteins may have arisen from a single common ancestor. Several hypothetical proteins, mainly of plant origin, that clearly contain the conserved Sm motifs but exhibit only comparatively low overall homology to one of the human Sm proteins, were identified in the data base. This suggests that the Sm motifs may also be shared by non-spliceosomal proteins. Further, we provide experimental evidence that the Sm motifs are involved, at least in part, in Sm protein-protein interactions. Specifically, we show by co-immunoprecipitation analyses of in vitro translated B' and D3 that the Sm motifs are essential for complex formation between B' and D3. Our finding that the Sm proteins share conserved sequence motifs may help to explain the frequent occurrence in patient sera of anti-Sm antibodies that cross-react with multiple Sm proteins and may ultimately further our understanding of how the snRNPs act as auto-antigens and immunogens in SLE.     

Binding of antibodies against high and low molecular weight cytokeratin proteins in the human placenta with special reference to infarcts, proliferation and differentiation processes

Recent immunocytochemical studies have shown that placental villous trophoblasts contain the high molecular weight cytokeratin (CK) proteins 5/6 and 17. In the case of CK 17, trophoblastic immunostaining was positive in villi covered by fibrinoid. CKs 5/6 and 17 are expressed by hyperproliferative cells. The aim of this investigation was to examine the location of these CKs in placental infarcts, known to be demarcated by fibrinoid and hyperproliferative trophoblasts. The results were compared with those obtained by immunostaining against Ki-67, tenascin and α1-, α6- and β1- integrins, which are involved in cell proliferation, differentiation and regenerative processes. Furthermore, the expression of the single CKs 7, 8, 10, 13, 14, 18 and 19 was investigated by immunocytochemistry and immunoblotting. While low and high molecular weight CKs were present in villous and extravillous trophoblasts, only low molecular weight CKs were detected in vascular and extravascular placental smooth muscle cells. Placental infarcts revealed different immunoreactivities in the infarct margin and centre: high molecular CKs, tenascin, Ki-67 and oncofoetal fibronectin predominated in the infarct margin, low molecular CKs, fibrin and integrins in the centre. The expression of tenascin and a defined change in the expression of CK 17 indicates villous repair and hyperproliferative mechanisms in placental infarcts. This revised version was published online in November 2006 with corrections to the Cover Date.     

Determination of apoplastic K+ in intact leaves by ratio imaging of PBFI fluorescence

The tetraammonium salt of the K⁺ binding fluorescent dye benzofuranisophthalate (PBFI) was used to investigate the influence ofpotassium nutrition (0.1–2.1 mol m^–3) on apoplasticK⁺ inVicia faba leaves by means of ratio imaging. As a referencethe infiltration-centrifugation method was used. Both methodsreflected the influence of K⁺ supply on apoplastic K⁺ concentration.The abaxial leaf side revealed significantly higher K⁺ concentrations(20-25 mol m^–3) than the adaxial side (5–8 mol m^–3).Application of CCCP led to an immediate increase in apoplasticK⁺ demonstrating the reliability of the PBFI method. Key words: Vicia faba, leaf, apoplast, K⁺, PBFI, ratio imaging, ratiometric fluorescence microscopy     

Ethanol Activates Maxi Ca2+-activated K+ Channels of Clonal Pituitary (GH3) Cells

The effect of ethanol on maxi Ca²⁺-activated K⁺ channels (BK channels) in GH3 pituitary tumor cells was investigated using single-channel recordings and focusing on intracellular signal transduction. In outside-out patches, ethanol caused a transient concentration-dependent increase of BK-channel activity. 30 mm (1.4‰) ethanol significantly increased mean channel open time and channel open probability by 26.3 ± 9% and 78.8 ± 10%, respectively; single-channel current amplitude was not affected by ethanol. The augmenting effect of ethanol was blocked in the presence of protein kinase C (PKC) inhibitors staurosporine, bisindolylmaleimide, and PKC (19–31) pseudosubstrate inhibitor as well as by AMP-PNP (5′-adenylylimidodiphosphate), a nonhydrolyzable ATP-analogue, but not by the phospholipase C blocker U-73122. Phosphatase inhibitors microcystin-LR and okadaic acid promoted the ethanol effect. The blocking effect was released at higher concentrations of ethanol (100 mm) suggesting a second site of action or a competition between blockers and ethanol. Our results suggest that the effect of ethanol on BK-channels is mediated by PKC stimulation and phosphorylation of the channels which increases channel activity and hence may influence action potentials duration and hormone secretion. Received: 24 July 1996/Revised: 27 December 1996