Sterols as dietary markers for Drosophila melanogaster.

During cold acclimation fruit flies switch their feeding from yeast to plant food, however there are no robust molecular markers to monitor this in the wild. Drosophila melanogaster is a sterol auxotroph and relies on dietary sterols to produce lipid membranes, lipoproteins and molting hormones. We employed shotgun lipidomics to quantify eight major food sterols in total lipid extracts of heads and genital tracts of adult male and female flies. We found that their sterol composition is dynamic and reflective of fly diet in an organ-specific manner. Season-dependent changes observed in the organs of wild-living flies suggested that the molar ratio between yeast (ergosterol, zymosterol) and plant (sitosterol, stigmasterol) sterols is a quantifiable, generic and unequivocal marker of their feeding behavior suitable for ecological and environmental population-based studies. The enrichment of phytosterols over yeast sterols in wild-living flies at low temperatures is consistent with switching from yeast to plant diet and corroborates the concomitantly increased unsaturation of their membrane lipids

Vergleichende genetische Kartierung der Feuerbrandresistenz bei Malus sp. = Comparative genetic mapping of resistance to fire blight in Malus sp.

Für die vergleichende Kartierung von Feuerbrandresistenz bei Malus wurden vier Populationen bearbeitet, die als einen Elter einen Donor für Feuerbrandresistenz und als zweiten Elter die anfällige Sorte ‘Idared’ hatten. Als Donoren für Widerstandsfähigkeit wurden definierte Abstammungen der Wildarten M. baccata, M. fusca, M. ×robusta und die widerstandsfähige Pillnitzer Apfelsorte ‘Rewena’ genutzt. Die Phänotypisierung der Populationen erfolgte in unterschiedlichen Jahren durch Inokulation von vegetativen Trieben mit dem Erwinia amylovora-Isolat Ea222 und anderen definierten Isolaten unter Gewächshausbedingungen. Der Befall der Triebe wurde als Befallsrate, dem Verhältnis der befallenen Trieblänge zur Gesamttrieblänge, ermittelt. Je nach Isolat und Population variierte die durchschnittliche Befallsrate zwischen 10 bis 82%. Die Genotypisierung der Nachkommen zur Erstellung genetischer Karten erfolgte mit Mikrosatelliten-, SNP- und DArT-Markern. Die genetischen Kopplungskarten wurden mittels Joinmap 4.0 und die Resistenz gegenüber Feuerbrand mit MapQTL5.0 kartiert. Major-QTLs konnten für M. ×robusta auf Kopplungsgruppe 3, für M. baccata auf Kopplungsgruppe 12 und für M. fusca auf Kopplungsgruppe 10 detektiert werden. Während die Feuerbrand-QTLs von M. ×robusta 5 und M. fusca je nach Jahr bis zu ca. 87% der phänotypischen Varianz erklären können, erklärt der QTL von M. baccata nur ca. 45%, was die Existenz eines zweiten QTL nahe legt. Für die resistente Sorte ‘Rewena’ konnte kein QTL detektiert werden. Mit den Markern, die im Bereich der detektierten QTLs liegen, kann auf Resistenz gegenüber Feuerbrand selektiert werden. Damit ist eine Kombinierung unterschiedlicher Feuerbrandresistenz-QTLs möglich. Auf dieser Basis können zukünftig Sorten gezüchtet werden, die dauerhaft widerstandsfähig gegenüber Feuerbrand sind, um diese dann dem deutschen Obstbau zur Verfügung stellen zu könne

Precocious expression of Blimp1 in B cells causes autoimmune disease with increased self-reactive plasma cells

The transcription factor Blimp1 is not only an essential regulator of plasma cells, but also a risk factor for the development of autoimmune disease in humans. Here, we demonstrate in the mouse that the Prdm1 (Blimp1) gene was partially activated at the chromatin and transcription level in early B cell development, although mature Prdm1 mRNA did not accumulate due to posttranscriptional regulation. By analyzing a mouse model that facilitated ectopic Blimp1 protein expression throughout B lymphopoiesis, we could demonstrate that Blimp1 impaired B cell development by interfering with the B cell gene expression program, while leading to an increased abundance of plasma cells by promoting premature plasmablast differentiation of immature and mature B cells. With progressing age, these mice developed an autoimmune disease characterized by the presence of autoantibodies and glomerulonephritis. Hence, these data identified ectopic Blimp1 expression as a novel mechanism, through which Blimp1 can act as a risk factor in the development of autoimmune disease

Precocious expression of Blimp1 in B cells causes autoimmune disease with increased self-reactive plasma cells

The transcription factor Blimp1 is not only an essential regulator of plasma cells, but also a risk factor for the development of autoimmune disease in humans. Here, we demonstrate in the mouse that the Prdm1 (Blimp1) gene was partially activated at the chromatin and transcription level in early B cell development, although mature Prdm1 mRNA did not accumulate due to posttranscriptional regulation. By analyzing a mouse model that facilitated ectopic Blimp1 protein expression throughout B lymphopoiesis, we could demonstrate that Blimp1 impaired B cell development by interfering with the B cell gene expression program, while leading to an increased abundance of plasma cells by promoting premature plasmablast differentiation of immature and mature B cells. With progressing age, these mice developed an autoimmune disease characterized by the presence of autoantibodies and glomerulonephritis. Hence, these data identified ectopic Blimp1 expression as a novel mechanism, through which Blimp1 can act as a risk factor in the development of autoimmune disease

CD22 ligand-binding and signaling domains reciprocally regulate B-cell Ca²⁺ signaling

A high proportion of human B cells carry B-cell receptors (BCRs) that are autoreactive. Inhibitory receptors such as CD22 can down-modulate autoreactive BCR responses. With its extracellular domain, CD22 binds to sialic acids in α2,6 linkages in cis, on the surface of the same B cell or in trans, on other cells. Sialic acids are self ligands, as they are abundant in vertebrates, but are usually not expressed by pathogens. We show that cis-ligand binding of CD22 is crucial for the regulation of B-cell Ca2+ signaling by controling the CD22 assocation to the BCR. Mice with a mutated CD22 ligand-binding domain of CD22 showed strongly reduced Ca2+ signaling. In contrast, mice with mutated CD22 immunoreceptor tyrsoine-based inhibition motifs have increased B-cell Ca2+ responses, increased B-cell turnover, and impaired survival of the B cells. Thus, the CD22 ligand-binding domain has a crucial function in regulating BCR signaling, which is relevant for controlling autoimmunity

Essential role for the transcription factor Bhlhe41 in regulating the development, self-renewal and BCR repertoire of B-1a cells

Innate-like B-1a cells provide a first line of defense against pathogens, yet little is known about their transcriptional control. Here we identified an essential role for the transcription factor Bhlhe41, with a lesser contribution by Bhlhe40, in controlling B-1a cell differentiation. Bhlhe41-/-Bhlhe40-/- B-1a cells were present at much lower abundance than were their wild-type counterparts. Mutant B-1a cells exhibited an abnormal cell-surface phenotype and altered B cell receptor (BCR) repertoire exemplified by loss of the phosphatidylcholine-specific VH12Vκ4 BCR. Expression of a pre-rearranged VH12Vκ4 BCR failed to 'rescue' the mutant phenotype and revealed enhanced proliferation accompanied by increased cell death. Bhlhe41 directly repressed the expression of cell-cycle regulators and inhibitors of BCR signaling while enabling pro-survival cytokine signaling. Thus, Bhlhe41 controls the development, BCR repertoire and self-renewal of B-1a cells

Host resistance to Erwinia amylovora: germplasm, breeding, genetics

The most important bacterial disease affecting pome fruit is fire blight caused by Erwinia amylovora. It can cause devastating economic losses and is reliably controlled only by the application of antibiotics, which are banned in many European countries due to environmental, sustainable and consumer friendly issues. One solution could be the utilization of fire blight resistant cultivars in apple production. In 2003, we started an approach at Dresden-Pillnitz to detect different mechanisms conferring resistance to fire blight aimed at their combination in new cultivars. Four segregating populations were established to map QTLs for fire blight resistance. The donors used were three wild species accessions Malus baccata (MALD0004), M. fusca (MALD0045), M. × robusta 5 and the Pillnitz cultivar Rewena. The susceptible parent in each case was Idared. Grafted scions of each progeny were inoculated with E. amylovora strain Ea 222_JKI in at least for two years. Average percent lesion length (PLL) of all progenies was determined. Genetic linkage maps were established using DArT-, SCAR-, SNP-, and SSR-markers. Whereas in Rewena no QTL could be determined, major QTLs were detected in M. baccata on linkage group 12, in M. fusca on linkage group 10, and in M. × robusta 5 on linkage group 3 explaining up to around 50, 85 and 85% of the phenotypic variance, respectively. The case that all resistance QTLs are located on different linkage groups enhances the chance that different mechanisms are acting in the donors. Additionally, trees of the Idared by M. × robusta 5 population were planted in an orchard and flowers were inoculated in two consecutive years. The QTL on linkage group 3 could be confirmed after mappin

QTL mapping for resistance to fire blight using several Erwinia amylovora strains resulting in different host-pathogen interactions

QTL mapping of fire blight resistance in apple is an effective tool to determine associations between regions in the genome of apple and resistance to the bacterial disease. Several wild Malus accessions of different apple species have been identified as resistant to fire blight making them valuable as donors for the introgression of resistance to the cultivated apple Malus × domestica Borkh. Resistant accessions of the wild species M. baccata, M. fusca and M. × robusta 5 (Mr5) were inoculated with the wild type strain Ea1189 and the AvrRpt2EA deletion mutant (pZYRKD3-1). While M. baccata and M. fusca showed no symptoms to pZYRKD3-1, the resistance of M. × robusta 5 was overcome by pZYRKD3-1 with an average necrosis length of 52% respectively. Inoculation of the mapping population ‘Idared’ × Mr5 with the strain Ea 1189 results in the confirmation of the QTL on LG 3 in Mr5; this QTL completely broke down after inoculation with pZYRKD3-1, but two minor QTLs on LG 7 and LG 11 were detected

Mapping of fire blight resistance in Malus ×robusta 5 flowers following artificial inoculation

Background Although the most common path of infection for fire blight, a severe bacterial disease on apple, is via host plant flowers, quantitative trait loci (QTLs) for fire blight resistance to date have exclusively been mapped following shoot inoculation. It is not known whether the same mechanism underlies flower and shoot resistance. Results We report the detection of a fire blight resistance QTL following independent artificial inoculation of flowers and shoots on two F1 segregating populations derived from crossing resistant Malus ×robusta 5 (Mr5) with susceptible ‘Idared’ and ‘Royal Gala’ in experimental orchards in Germany and New Zealand, respectively. QTL mapping of phenotypic datasets from artificial flower inoculation of the ‘Idared’ × Mr5 population with Erwinia amylovora over several years, and of the ‘Royal Gala’ × Mr5 population in a single year, revealed a single major QTL controlling floral fire blight resistance on linkage group 3 (LG3) of Mr5. This QTL corresponds to the QTL on LG3 reported previously for the ‘Idared’ × Mr5 and an ‘M9’ × Mr5 population following shoot inoculation in the glasshouse. Interval mapping of phenotypic data from shoot inoculations of subsets from both flower resistance populations re-confirmed that the resistance QTL is in the same position on LG3 of Mr5 as that for flower inoculation. These results provide strong evidence that fire blight resistance in Mr5 is controlled by a major QTL on LG3, independently of the mode of infection, rootstock and environment. Conclusions This study demonstrates for the first time that resistance to fire blight caused by Erwinia amylovora is independent of the mode of inoculation at least in Malus ×robusta 5