Durch Phytochrom kontrollierte Enzyme der Flavonoidsynthese in Zellsuspensionskulturen von Petersilie (Petroselinum hortense Hoffm.)

Summary Enzymes involved in flavonoid synthesis in parsley (Petroselinum nortense Hoffm.) cell suspension cultures are induced by light. It has been suggested that the regulation of the enzymes of phenylpropanoid metabolism (group 1) is independent of that of the enzymes of flavonoid synthesis proper (group 2) [Hahl-brock, K., Ebel, J., Ortmann, R., Sutter, A., Wellmann, E., Grisebach, H.: Biochim. biophys. Acta (Amst.) 244, 7–15 (1971)]. Phytochrome was demonstrated to be involved in the light effect controlling flavonoid synthesis. Phytochrome is only effective after a preceding irradiation with ultraviolet light (_max<300 nm) [Wellmann, E.: Planta (Berl.) 101, 283–286 (1971)]. — In order to determine whether phytochrome affects phenylalanine ammonia-lyase (PAL; EC 4.3.1.5), a group 1 enzyme, or exerts a general effect of all enzymes of the flavonoid pathway. PAL and two enzymes from group 2, chalcone-flavanone isomerase and UDP-apiose synthetase, were investigated. Under appropriate conditions of irradiation (low UV-dose with subsequent red/far-red pulses) both group 2 enzymes were shown to be controlled by phytochrome to the same extent as PAL. The UV-induced increase in activity of all three enzymes is reduced to about 70% by an irradiation for 10 min with far-red light. This far-red effect is fully reversible by a subsequent irradiation with 10 mith red light. There is no evidence for a rate limiting enzyme (or group of enzymes) involved in flavonoid formation with respect to phytochrome control.

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Abkürzungen PAL Phenylalaninammoniumlyasc - CFI Chalkonflavanonisomerase     

Galápagos and Californian sea lions are separate species: Genetic analysis of the genus Zalophus and its implications for conservation management

The climatic cycles with subsequent glacial and intergalcial periods have had a great impact on the distribution and evolution of species. Using genetic analytical tools considerably increased our understanding of these processes. In this review I therefore give an overview of the molecular biogeography of Europe. For means of simplification, I distinguish between three major biogeographical entities: (i) "Mediterranean" with Mediterranean differentiation and dispersal centres, (ii) "Continental" with extra-Mediterranean centres and (iii) "Alpine" and/or "Arctic" with recent alpine and/or arctic distribution patterns. These different molecular biogeographical patterns are presented using actual examples. Many "Mediterranean" species are differentiated into three major European genetic lineages, which are due to glacial isolation in the three major Mediterranean peninsulas. Postglacial expansion in this group of species is mostly influenced by the barriers of the Pyrenees and the Alps with four resulting main patterns of postglacial range expansions. However, some cases are known with less than one genetic lineage per Mediterranean peninsula on the one hand, and others with a considerable genetic substructure within each of the Mediterranean peninsulas, Asia Minor and the Maghreb. These structures within the Mediterranean sub-centres are often rather strong and in several cases even predate the Pleistocene. For the "Continental" species, it could be shown that the formerly supposed postglacial spread from eastern Palearctic expansion centres is mostly not applicable. Quite the contrary, most of these species apparently had extra-Mediterranean centres of survival in Europe with special importance of the perialpine regions, the Carpathian Basin and parts of the Balkan Peninsula. In the group of "Alpine" and/or "Arctic" species, several molecular biogeographical patterns have been found, which support and improve the postulates based on distribution patterns and pollen records. Thus, genetic studies support the strong linkage between southwestern Alps and Pyrenees, northeastern Alps and Carpathians as well as southeastern Alps and the Dinaric mountain systems, hereby allowing conclusions on the glacial distribution patterns of these species. Furthermore, genetic analyses of arctic-alpine disjunct species support their broad distribution in the periglacial areas at least during the last glacial period. The detailed understanding of the different phylogeographical structures is essential for the management of the different evolutionary significant units of species and the conservation of their entire genetic diversity. Furthermore, the distribution of genetic diversity due to biogeographical reasons helps understanding the differing regional vulnerabilities of extant populations.     

An evaluation of LSU rDNA D1-D2 sequences for their use in species identification

Background

The Radical Pair model proposes that magnetoreception is a light-dependent process. Under low monochromatic light from the short-wavelength part of the visual spectrum, migratory birds show orientation in their migratory direction. Under monochromatic light of higher intensity, however, they showed unusual preferences for other directions or axial preferences. To determine whether or not these responses are still controlled by the respective light regimes, European robins, Erithacus rubecula, were tested under UV, Blue, Turquoise and Green light at increasing intensities, with orientation in migratory direction serving as a criterion whether or not magnetoreception works in the normal way.

Results

The birds were well oriented in their seasonally appropriate migratory direction under 424 nm Blue, 502 nm Turquoise and 565 nm Green light of low intensity with a quantal flux of 8·10¹⁵ quanta s^-1 m^-2, indicating unimpaired magnetoreception. Under 373 nm UV of the same quantal flux, they were not oriented in migratory direction, showing a preference for the east-west axis instead, but they were well oriented in migratory direction under UV of lower intensity. Intensities of above 36·10¹⁵ quanta s^-1 m^-2 of Blue, Turquoise and Green light elicited a variety of responses: disorientation, headings along the east-west axis, headings along the north-south axis or 'fixed' direction tendencies. These responses changed as the intensity was increased from 36·10¹⁵ quanta s^-1 m^-2 to 54 and 72·10¹⁵ quanta s^-1 m^-2.

Conclusion

The specific manifestation of responses in directions other than the migratory direction clearly depends on the ambient light regime. This implies that even when the mechanisms normally providing magnetic compass information seem disrupted, processes that are activated by light still control the behavior. It suggests complex interactions between different types of receptors, magnetic and visual. The nature of the receptors involved and details of their connections are not yet known; however, a role of the color cones in the processes mediating magnetic input is suggested.     

Parallel selection mapping using artificially selected mice reveals body weight control loci

Understanding how polygenic traits evolve under selection is an unsolved problem, because challenges exist for identifying genes underlying a complex trait and understanding how multilocus selection operates in the genome. Here we study polygenic response to selection using artificial selection experiments. Inbred strains from seven independent long-term selection experiments for extreme mouse body weight ("high" lines weigh 42-77 g versus 16-40 g in "control" lines) were genotyped at 527,572 SNPs to identify loci controlling body weight. We identified 67 parallel selected regions (PSRs) where high lines share variants rarely found among the controls. By comparing allele frequencies in one selection experiment against its unselected control, we found classical selective sweeps centered on the PSRs. We present evidence supporting two G protein-coupled receptors GPR133 and Prlhr as positional candidates controlling body weight. Artificial selection may mimic natural selection in the wild: compared to control loci, we detected reduced heterozygosity in PSRs in unusually large wild mice on islands. Many PSRs overlap loci associated with human height variation, possibly through evolutionary conserved functional pathways. Our data suggest that parallel selection on complex traits may evoke parallel responses at many genes involved in diverse but relevant pathways.     

her1 and her13.2 are jointly required for somitic border specification along the entire axis of the fish embryo

Delta-Notch and FGF signaling are involved in the control of somitogenesis in zebrafish. her genes are generally known as downstream targets of Delta-Notch signaling, but the her13.2 gene from zebrafish has recently been shown to depend on FGF signaling only. We have here studied the functional role of her13.2 in conjunction with her genes that are under Delta-Notch control. We show that joint inactivation of her1 and her13.2 leads to a complete loss of all somitic borders, including the most anterior ones. This somitic phenotype is much stronger than would be expected from the effects of the inactivation of either gene alone. A joint inactivation of her13.2 and her7, which is a paralogue of her1, does not show this enhanced effect. Thus, our results confirm inferences from in vitro studies that her1 and her13.2 form specific heterodimers, which may directly be required for regulating further target genes. These two her genes thus constitute the link between Delta-Notch pathway and FGF signaling during entire somitogenesis. We show that this interaction is conserved in the rice fish medaka, as a joint inactivation of the respective orthologues leads also to the same phenotype as in zebrafish. In addition, our results suggest that the mechanisms for anterior and posterior somite formation are not principally different, although the anterior somites often seem more refractory to genetic perturbations.     

Genome‐scale analysis of library sorting (GALibSo): Isolation of secretion enhancing factors for recombinant protein production in Pichia pastoris

A method combining fluorescence activated cell sorting (FACS) and DNA microarray assisted clone identification was developed and termed Genome‐Scale Analysis of Library Sorting (GALibSo). Genes enhancing the production of secreted heterologous proteins in Pichia pastoris were identified out of a cDNA library by cell surface display and FACS. The trends of gene enrichment during consecutive FACS rounds were monitored by DNA microarrays. In a case study a P. pastoris cDNA library was co‐expressed in a strain secreting the Fab fragment of a monoclonal antibody against human immunodeficiency virus type 1 as a model protein. Three genes were identified, increasing the relative expression level of the surface‐displayed model protein up to 45%. While one of these genes had a positive effect on three out of four tested proteins, the product specific effect of the other two suggested that the effects of the co‐expressed secretion enhancing factors are partly dependent on the protein to be produced. The microarray based monitoring of the enrichment of genes causing enhanced protein secretory capacity led to novel insights into the limitation of protein secretion. Biotechnol. Bioeng. 2010; 105: 543–555. © 2009 Wiley Periodicals, Inc.     

Engineering of biotin-prototrophy in Pichia pastoris for robust production processes

Biotin plays an essential role as cofactor for biotin-dependent carboxylases involved in essential metabolic pathways. The cultivation of Pichia pastoris, a methylotrophic yeast that is successfully used as host for the production of recombinant proteins, requires addition of high dosage of biotin. As biotin is the only non-salt media component used during P. pastoris fermentation (apart from the carbon source), nonconformities during protein production processes are usually attributed to poor quality of the added biotin.In order to avoid dismissed production runs due to biotin quality issues, we engineered the biotin-requiring yeast P. pastoris to become a biotin-prototrophic yeast. Integration of four genes involved in the biotin biosynthesis from brewing yeast into the P. pastoris genome rendered P. pastoris biotin-prototrophic. The engineered strain has successfully been used as production host for both intracellular and secreted heterologous proteins in fed-batch processes, employing mineral media without vitamins. Another field of application for these truly prototrophic hosts is the production of biochemicals and small metabolites, where defined mineral media leads to easier purification procedures.