Non-targeted metabolic profiling of BW312 Hordeum vulgare semi dwarf mutant using UHPLC coupled to QTOF high resolution mass spectrometry

Abstract Barley (Hordeum vulgare) is the fourth crop cultivated in the world for human consumption and animal feed, making it important to breed healthy and productive plants. Among the threats for barley are lodging, diseases, and pathogens. To avoid lodging, dwarf and semi-dwarf mutants have been selected through breeding processes. Most of these mutants are affected on hormonal biosynthesis or signalling. Here, we present the metabolic characterization of a brassinosteroid insensitive semi-dwarf mutant, BW312. The hormone profile was determined through a targeted metabolomics analysis by UHPLC-triple quadrupole-MS/MS, showing an induction of gibberellic acid and jasmonic acid in the semi-dwarf mutant. A non-targeted metabolomics analysis by UHPLC-QTOF-MS/MS revealed a differential metabolic profile, with 16 and 9 metabolites showing higher intensities in the mutant and wild-type plants respectively. Among these metabolites, azelaic acid was identified. Gibberellic acid, jasmonic acid, and azelaic acid are involved in pathogen resistance, showing that this semi-dwarf line has an enhanced basal pathogen resistance in absence of pathogens, and therefore is of interest in breeding programs to fight against lodging, but also probably to increase pathogen resistance

Enolases: Storage compounds in seeds? Evidence from a proteomic comparison of zygotic and somatic embryos of Cyclamen persicum Mill

Somatic embryogenesis is well established for the economic relevant ornamental crop Cyclamen and thus could supplement the elaborate propagation via seeds. However, the use of somatic embryogenesis for commercial large scale propagation is still limited due to physiological disorders and asynchronous development within emerged embryos. To overcome these problems, profound knowledge of the physiological processes in Cyclamen embryogenesis is essential. Thus, the proteomes of somatic and zygotic embryos were characterised in a comparative approach. Protein separation via two dimensional IEF-SDS PAGE led to a resolution of more than 1,000 protein spots/gel. Overall, 246 proteins were of differential abundance in the two tissues compared. Mass spectrometry analysis of the 300 most abundant protein spots resulted in the identification of 247 proteins, which represent 90 distinct protein species. Fifty-five percent of the 247 proteins belong to only three physiological categories: glycolysis, protein folding and stress response. The latter physiological process was especially predominant in the somatic embryos. Remarkably, the glycolytic enzyme enolase was the protein most frequently detected and thus is supposed to play an important role in Cyclamen embryogenesis. Data are presented that indicate involvement of "small enolases" as storage proteins in Cyclamen. A digital reference map was established via a novel software tool for the web-based presentation of proteome data linked to KEGG and ExPasy protein-databases and both were made publicly available online. © 2011 Springer Science+Business Media B.V

Early manganese-toxicity response in Vigna unguiculata L. - A proteomic and transcriptomic study

The apoplast is known to play a predominant role in the expression of manganese (Mn) toxicity in cowpea (Vigna unguiculata L.) leaves. To unravel early Mn-toxicity responses after 1-3 days Mn treatment also in the leaf symplast, we studied the symplastic reactions induced by Mn in two cultivars differing in Mn tolerance on a total cellular level. Comparative proteome analyses of plants exposed to low or high Mn allowed to identify proteins specifically affected by Mn, particularly in the Mn-sensitive cowpea cultivar. These proteins are involved in CO2 fixation, stabilization of the Mn cluster of the photosystem II, pathogenesis-response reactions and protein degradation. Chloroplastic proteins important for CO2 fixation and photosynthesis were of lower abundance upon Mn stress suggesting scavenging of metabolic energy for a specific stress response. Transcriptome analyses supported these findings, but additionally revealed an upregulation of genes involved in signal transduction only in the Mn-sensitive cultivar. In conclusion, a coordinated interplay of apoplastic and symplastic reactions seems to be important during the Mn-stress response in cowpea. © 2008 Wiley-VCH Verlag GmbH & Co. KGaA

Vitamin D5 in Arabidopsis thaliana

Abstract Vitamin D3 is a secosterol hormone critical for bone growth and calcium homeostasis, produced in vertebrate skin by photolytic conversion of the cholesterol biosynthetic intermediate provitamin D3. Insufficient levels of vitamin D3 especially in the case of low solar UV-B irradiation is often compensated by an intake of a dietary source of vitamin D3 of animal origin. Small amounts of vitamin D3 were described in a few plant species and considered as a peculiar feature of their phytochemical diversity. In this report we show the presence of vitamin D5 in the model plant Arabidopsis thaliana. This plant secosterol is a UV-B mediated derivative of provitamin D5, the precursor of sitosterol. The present work will allow a further survey of vitamin D distribution in plant species

Social policies, separation and second birth spacing in Western Europe

The research leading to these results received funding from the European Union’s Seventh Framework Programme (FP7/2007–2013) under grant agreement no. 320116 for the research project FamiliesAndSocieties. We also acknowledge funds from the Academy of Finland (decision number 275030 and 293103) and the German Science Foundation (KR 2855/3–1 and TR 457/7–1).Objective: This paper studies postseparation fertility behavior. The aim is to investigate whether, and if so how, separation affects second birth spacing in Western European countries. Methods: This analysis makes use of rich survey data from Belgium, France, Germany, Italy, Spain, and the United Kingdom, as well as from Finnish register data. We thus cover the behavior of a large proportion of the population of Western Europe. We also use descriptive measures, such as Kaplan‒Meier survival functions and cumulative incidence curves. In the multivariate analysis, we employ event history modeling to show how education relates to postseparation fertility behavior. Results: There are large differences in postseparation fertility behavior across European countries. For Spain and Italy, we find that only a negligibly small proportion of the population have a second child after separating from the other parent of the firstborn child. The countries with the highest proportion of second children with a new partner are the United Kingdom, Germany, and Finland. In all countries, separation after first birth leads to a sharp increase in the birth interval between first and second births. Contribution: Our study is a contribution to the demographic literature that aims at understanding birth spacing patterns in Western Europe. Furthermore, we draw attention to the role of postseparation policies in explaining country differences in fertility behavior in contemporary societies.Publisher PDFPeer reviewe

Proteomic and phosphoproteomic analysis of polyethylene glycol-induced osmotic stress in root tips of common bean (Phaseolus vulgaris L.)

Previous studies have shown that polyethylene glycol (PEG)-induced osmotic stress (OS) reduces cell-wall (CW) porosity and limits aluminium (Al) uptake by root tips of common bean (Phaseolus vulgaris L.). A subsequent transcriptomic study suggested that genes related to CW processes are involved in adjustment to OS. In this study, a proteomic and phosphoproteomic approach was applied to identify OS-induced protein regulation to further improve our understanding of how OS affects Al accumulation. Analysis of total soluble proteins in root tips indicated that, in total, 22 proteins were differentially regulated by OS; these proteins were functionally categorized. Seventy-seven per- cent of the total expressed proteins were involved in metabolic pathways, particularly of carbohydrate and amino acid metabolism. An analysis of the apoplastic proteome revealed that OS reduced the level of five proteins and increased that of seven proteins. Investigation of the total soluble phosphoproteome suggested that dehydrin responded to OS with an enhanced phosphorylation state without a change in abundance. A cellular immunolocalization analysis indicated that dehydrin was localized mainly in the CW. This suggests that dehydrin may play a major protective role in the OS-induced physical breakdown of the CW structure and thus maintenance of the reversibility of CW extensibility during recovery from OS. The proteomic and phosphoproteomic analyses provided novel insights into the complex mechanisms of OS-induced reduction of Al accumulation in the root tips of common bean and highlight a key role for modification of CW structure.BMZ/GTZ/05.7860.9-001.00BMZ/GTZ/05.7860.9-001.0

Hibernating brown bears are protected against atherogenic dyslipidemia

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Phospholipid Scramblase-1-Induced Lipid Reorganization Regulates Compensatory Endocytosis in Neuroendocrine Cells

Calcium-regulated exocytosis in neuroendocrine cells and neurons is accompanied by the redistribution of phosphatidylserine (PS) to the extracellular space, leading to a disruption of plasma membrane asymmetry. How and why outward translocation of PS occurs during secretion are currently unknown. Immunogold labeling on plasma membrane sheets coupled with hierarchical clustering analysis demonstrate that PS translocation occurs at the vicinity of the secretory granule fusion sites. We found that altering the function of the phospholipid scramblase-1 (PLSCR-1) by expressing a PLSCR-1 calcium-insensitive mutant or by using chromaffin cells from PLSCR-1−/−mice prevents outward translocation of PS in cells stimulated for exocytosis. Remarkably, whereas transmitter release was not affected, secretory granule membrane recapture after exocytosis was impaired, indicating that PLSCR-1 is required for compensatory endocytosis but not for exocytosis. Our results provide the first evidence for a role of specific lipid reorganization and calcium-dependent PLSCR-1 activity in neuroendocrine compensatory endocytosis