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Die Referentin erläutert den Ursprung embryonaler Stammzellen und woher sie kommen. Gezeigt wird weiter unter anderem die Gewinnung embryonaler Stammzellen aus der Blastozyste der Maus, und es wird auf die Differenzierung von glatter Muskulatur und Knochen eingegangen. Die Wissenschaftlerin sieht in der Etablierung neuer in vitro Modelle eine große Perspektive für die Forschung mit Maus-Stammzellen.
Die vorliegende Arbeit wurde im Zeitraum von Oktober 2002 bis November 2005 an dem Institut für Biochemie und Biologie der Universität Potsdam in Kooperation mit dem Institut für Chemie des GKSS Forschungszentrums in Teltow unter der Leitung von Herrn Prof. Dr. B. Micheel und Herrn Prof. Dr. Th. Groth angefertigt. Im Rahmen dieser Arbeit wurden die Wechselwirkungen von Immunzellen mit verschiedenen Kultursubstraten untersucht. Dafür wurden drei verschiedene Hybridomzelllinien eingesetzt. Eine Hybridomzelllinie (K2) ist im Laufe dieser Arbeit hergestellt und etabliert worden. Der Einsatz von synthetischen und proteinbeschichteten Kulturoberflächen führte bei Hybridomzellen zu einer deutlich gesteigerten Antikörpersynthese im Vergleich zu herkömmlichen Zellkulturmaterialien. Obwohl diese Zellen in der Regel als Suspensionszellen kultiviert werden, führten die eingesetzten Polymermembranen (PAN, NVP) zu einer verbesserten Antikörpersynthese (um 30%) gegenüber Polystyrol als Referenz. Es konnte gezeigt werden, dass es einen Zusammenhang zwischen der Produktivität und dem Adh asionsverhalten der Hybridomzellen gibt. Um den Einfluss von Proteinen der extrazellulären Matrix auf Zellwachstum und Antikörpersynthese von Hybridomzellen zu untersuchen, wurden proteinbeschichtete Polystyrol-Oberflächen eingesetzt. Für die Modifikationen wurden Fibronektin, Kollagen I, Laminin und BSA ausgewählt. Die Modifikation der Polystyrol-Oberfläche mit geringen Mengen Fibronektin (0,2-0,4 µg/ml) führte zu einer beträchtlichen Steigerung der Antikörpersynthese um 70-120%. Für Kollagen I- und BSA-Beschichtungen konnten Steigerungen von 40% beobachtet werden. Modifikationen der Polystyrol-Oberfläche mit Laminin zeigten nur marginale Effekte. Durch weitere Versuche wurde bestätigt, dass die Adhäsion der Zellen an Kollagen I- und Laminin-beschichteten Oberflächen verringert ist. Die alpha2-Kette des alpha2beta1-Integrins konnte auf der Zelloberfläche nicht nachgewiesen werden. Durch ihr Fehlen wird wahrscheinlich die Bindungsfähigkeit der Zellen an Kollagen I und Laminin beeinflusst. Durch die Ergebnisse konnte gezeigt werden, dass Hybridomzellen nicht nur Suspensionszellen sind und das Kultursubstrate das Zellwachstum und die Produktivität dieser Zellen stark beeinflussen können. Der Einsatz von synthetischen und proteinbeschichteten Kultursubstraten zur Steigerung der Antikörpersynthese kann damit für die industrielle Anwendung von großer Relevanz sein. Für die Modellierung einer Lymphknotenmatrix wurden Fibronektin, Kollagen I, Heparansulfat und N-Acetylglucosamin-mannose in verschiedenen Kombinationen an Glasoberflächen adsorbiert und für Versuche zur In-vitro-Immunisierung eingesetzt. Es konnte gezeigt werden, dass die Modifikation der Oberflächen die Aktivierung und Interaktion von dendritischen Zellen, T- und B-Lymphozyten begünstigt, was durch den Nachweis spezifischer Interleukine (IL12, IL6) und durch die Synthese spezifischer Antikörper bestätigt wurde. Eine spezifische Immunreaktion gegen das Antigen Ovalbumin konnte mit den eingesetzten Zellpopulationen aus Ovalbumin-T-Zell-Rezeptor-transgenen Mäusen nachgewiesen werden. Die In-vitro-Immunantwort wurde dabei am stärksten durch eine Kombination von Kollagen I, Heparansulfat und N-Acetylglucosamin-mannose auf einer Glasoberfläche gefördert. Die Etablierung einer künstlichen Immunreaktion kann eine gesteuerte Aktivierung bzw. Inaktivierung von körpereigenen dendritischen Zellen gegen bestehende Krankheitsmerkmale in vitro ermöglichen. Durch die Versuche wurden Grundlagen für spezifische Immunantworten erarbeitet, die u.a. für die Herstellung von humanen Antikörpern eingesetzt werden können.
Weltweit versuchen Wissenschaftler, künstliche Viren für den Gentransfer zu konstruieren, die nicht reproduktionsfähig sind. Diese sollen die Vorteile der natürlichen Viren besitzen (effizienter Transport von genetischem Material), jedoch keine Antigene auf ihrer Oberfläche tragen, die Immunreaktionen auslösen. Ziel dieses Projektes ist es, einen künstlichen Viruspartikel herzustellen, dessen Basis eine Polyelektrolytenhohlkugel bildet, die mit einer Lipiddoppelschicht bedeckt ist. Um intakte Doppelschichten zu erzeugen, muss die Wechselwirkung zwischen Lipid und Polyelektrolyt (z.B. DNA) verstanden und optimiert werden. Dazu ist es notwendig, die strukturelle Grundlage der Interaktion aufzuklären. Positiv geladene Lipide gehen zwar starke Wechselwirkungen mit der negativ geladenen DNA ein, sie wirken jedoch toxisch auf biologische Zellen. In der vorliegenden Arbeit wurde daher die durch zweiwertige Kationen vermittelte Kopplung von genomischer oder Plasmid-DNA an zwitterionische oder negativ geladene Phospholipide an zwei Modellsystemen untersucht. 1. Modellsystem: Lipidmonoschicht an der Wasser/Luft-Grenzfläche Methoden: Filmwaagentechnik in Kombination mit IR-Spektroskopie (IRRAS), Röntgenreflexion (XR), Röntgendiffraktion (GIXD), Brewsterwinkel-Mikroskopie (BAM), Röntgenfluoreszenz (XRF) und Oberflächenpotentialmessungen Resultate: A) Die Anwesenheit der zweiwertigen Kationen Ba2+, Mg2+, Ca2+ oder Mn2+ in der Subphase hat keinen nachweisbaren Einfluss auf die Struktur der zwitterionischen DMPE- (1,2-Dimyristoyl-phosphatidyl-ethanolamin) Monoschicht. B) In der Subphase gelöste DNA adsorbiert nur in Gegenwart dieser Kationen an der DMPE-Monoschicht. C) Sowohl die Adsorption genomischer Kalbsthymus-DNA als auch der Plasmid-DNA pGL3 bewirkt eine Reduktion des Neigungswinkels der Alkylketten, die auf einen veränderten Platzbedarf der Kopfgruppe zurückzuführen ist. Durch die Umorientierung der Kopfgruppe wird die elektrostatische Wechselwirkung zwischen den positiv geladenen Stickstoffatomen der Lipidkopfgruppen und den negativ geladenen DNA-Phosphaten erhöht. D) Die adsorbierte DNA weist eine geordnete Struktur auf, wenn sie durch Barium-, Magnesium-, Calcium- oder Manganionen komplexiert ist. Der Abstand zwischen parallelen DNA-Strängen hängt dabei von der Größe der DNA-Fragmente sowie von der Art des Kations ab. Die größten Abstände ergeben sich mit Bariumionen, gefolgt von Magnesium- und Calciumionen. Die kleinsten DNA-Abstände werden durch Komplexierung mit Manganionen erhalten. Diese Ionenreihenfolge stellt sich sowohl für genomische DNA als auch für Plasmid-DNA ein. E) Die DNA-Abstände werden durch die Kompression des Lipidfilms nicht beeinflusst. Zwischen der Lipidmonoschicht und der adsorbierten DNA besteht demnach nur eine schwache Wechselwirkung. Offensichtlich befindet sich die durch zweiwertige Kationen komplexierte DNA als weitgehend eigenständige Schicht unter dem Lipidfilm. 2. Modellsystem: Lipiddoppelschicht an der fest/flüssig-Grenzfläche Methoden: Neutronenreflexion (NR) und Quarzmikrowaage (QCM-D) Resultate: A) Das zwitterionische Phospholipid DMPC (1,2-Dimyristoyl-phosphatidylcholin) bildet keine Lipiddoppelschicht auf planaren Polyelektrolytmultischichten aus, deren letzte Lage das positiv geladene PAH (Polyallylamin) ist. B) Hingegen bildet DMPC auf dem negativ geladenen PSS (Polystyrolsulfonat) eine Doppelschicht aus, die jedoch Defekte aufweist. C) Eine Adsorption von genomischer Kalbsthymus-DNA auf dieser Lipidschicht findet nur in Gegenwart von Calciumionen statt. Andere zweiwertige Kationen wurden nicht untersucht. D) Das negativ geladene Phospholipid DLPA (1,2-Dilauryl-phosphatidsäure) bildet auf dem positiv geladenen PAH eine Lipiddoppelschicht aus, die Defekte aufweist. E) DNA adsorbiert ebenfalls erst in Anwesenheit von Calciumionen in der Lösung an die DLPA-Schicht. F) Durch die Zugabe von EDTA (Ethylendiamintetraessigsäure) werden die Calciumionen dem DLPA/DNA-Komplex entzogen, wodurch dieser dissoziiert. Demnach ist die calciuminduzierte Bildung dieser Komplexe reversibel.
We compared growth rates and efficiencies of pelagic bacteria from an extremely acidic mining lake (pH 2.6, mean depth 4.6m) supplied with different sources of carbon: (1) excreted by phytoplankton, (2) derived from benthic algae, (3) entering the lake via ground water, and (4) leached from leaf litter. Bacteria exhibited high growth rate and efficiency on exudates of pelagic and benthic algae. In contrast, they showed a lower growth rate and efficiency with organic carbon from ground water, and grew at a very high rate but a very low efficiency on leaf leachate. Results from stable isotope analyses indicate a greater importance of benthic exudates and leaf leachate for bacteria in the epilimnion, and a higher impact of ground water sources in the hypolimnion. Given the magnitude of differential source inputs into the lake, we suggest that benthic primary production was the most important carbon source for pelagic bacteria. The benthic-pelagic coupling seems to be more relevant in this shallow acidic lake with low pelagic carbon dioxide concentrations than in neutral lakes
Use of large Acacia trees by the cavity dwelling Black-tailed Tree Rat in the southern Kalahari
(2006)
Recent extensive harvesting of large, often dead Acacia trees in and savanna of southern Africa is cause for concern about the conservation status of the arid savanna and its animal community. We mapped vegetation and nests of the Black-tailed Tree Rat Thallomy's nigricauda to assess the extent to which the rats depend on particular tree species and on the existence of dead, standing trees. The study was conducted in continuous Acacia woodland on the southern and eastern edge of the Kalahari, South Africa. Trees in which there were tree rat nests were compared with trees of similar size and vigour to identify the characteristics of nest sites. Spatial analysis of tree rat distribution was conducted using Ripley's-L function. We found that T nigricauda was able to utilize all available tree species, as long as trees were large and old enough so that cavities were existing inside the stem. The spatial distribution of nest trees did not show clumping at the investigated scale, and we therefore reject the notion of the rats forming colonies when inhabiting continuous woodlands. The selection of a particular tree as a nest site was furthermore depending on the close proximity of the major food plant, Acacia mellifera. This may limit the choice of suitable nest sites. since A. mellifera was less likely to grow within a vegetation patch containing a large trees than in patches without large trees.
Filamin c is the predominantly expressed filamin isoform in striated muscles. It is localized in myofibrillar Z- discs, where it binds FATZ and myotilin, and in myotendinous junctions and intercalated discs. Here, we identify Xin, the protein encoded by the human gene 'cardiomyopathy associated 1' (CMYA1) as filamin c binding partner at these specialized structures where the ends of myofibrils are attached to the sarcolemma. Xin directly binds the EVH1 domain proteins Mena and VASP. In the adult heart, Xin and Mena/VASP colocalize with filamin c in intercalated discs. In cultured cardiomyocytes, the proteins also localize in the nonstriated part of myofibrils, where sarcomeres are assembled and an extensive reorganization of the actin cytoskeleton occurs. Unusual intraexonic splicing events result in the existence of three Xin isoforms that associate differentially with its ligands. The identification of the complex filamin c-Xin-Mena/VASP provides a first glance on the role of Xin in the molecular mechanisms involved in developmental and adaptive remodeling of the actin cytoskeleton during cardiac morphogenesis and sarcomere assembly. (c) 2006 Elsevier Inc. All rights reserved
Investigating the mechanisms which underlie the biomass fluctuations of populations and communities is important to better understand the processes which buffer community biomass in a variable environment. Based on long- term data of plankton biomass in Lake Constance (Bodensee), this study aims at explaining the different degree of synchrony among populations observed within two freshwater plankton groups, phytoplankton and ciliates. Established measures of temporal variability such as the variance ratio and cross-correlation coefficients were combined with first- order autoregressive models that allow estimating species interactions from time-series data. We found that predation was an important driver of the observed seasonal variability patterns in phytoplankton and ciliates, and that competitive interactions only played a subordinate role. In Lake Constance copepods and cladocerans, two major invertebrate predator groups, focus their grazing pressure at different times of the season. Model results suggested that compensatory dynamics detected in phytoplankton originate from the differential vulnerability of species to either one of these two predator groups. For ciliates model results advocated that synchrony among species occurs because ciliates tend to be vulnerable to both predator groups. Our findings underline the necessity of extending studies of community variability to multiple trophic levels because accounting for predator-prey interactions may often be more important than accounting for competitive interactions at one trophic level
In Arabidopsis thaliana and Oryza sativa, two types of PI 4-kinase (PI4Ks) have been isolated and functionally characterized. The alpha-type PI4Ks (similar to 220 kDa) contain a PH domain, which is lacking in beta-type PI4Ks (similar to 120 kDa). beta-Type PI4Ks, exemplified by Arabidopsis AtPI4K beta and rice OsPI4K2, contain a highly charged repetitive segment designated PPC (Plant PI4K Charged) region, which is an unique domain only found in plant beta-type PI4Ks at present. The PPC region has a length of similar to 300 amino acids and harboring 11 (AtPI4K beta) and 14 (OsPI4K2) repeats, respectively, of a 20-aa motif. Studies employing a modified yeast-based "Sequence of Membrane- Targeting Detection'' system demonstrate that the PPC(OsPI4K2) region, as well as the former 8 and latter 6 repetitive motifs within the PPC region, are able to target fusion proteins to the plasma membrane. Further detection on the transiently expressed GFP fusion proteins in onion epidermal cells showed that the PPC(OsPI4K2) region alone, as well as the region containing repetitive motifs 1-8, was able to direct GFP to the plasma membrane, while the regions containing less repetitive motifs, i.e. 6, 4, 2 or single motif(s) led to predominantly intracellular localization. Agrobacterium-mediated transient expression of PPC-GFP fusion protein further confirms the membrane-targeting capacities of PPC region. In addition, the predominant plasma membrane localization of AtPI4Kb was mediated by the PPC region. Recombinant PPC peptide, expressed in E. coli, strongly binds phosphatidic acid, PI and PI4P, but not phosphatidylcholine, PI5P, or PI(4,5) P-2 in vitro, providing insights into potential mechanisms for regulating sub- cellular localization and lipid binding for the plant beta-type PI4Ks
The ancestral galectin from the sponge Geodia cydonium (GCG) is classified on a structural basis to the prototype subfamily, whereas its carbohydrate-binding specificity is related to that of the mammalian chimera-type galectin-3. This dual coordination reveals GCG as a potential precursor of the later evolved galectin subfamilies, which is reflected in the primary structure of the protein. This study provides evidence that GCG is the LECT1 gene product, while neither a previously described LECT2 gene nor a functional LECT2 gene product was found in the specimen under investigation. The electrophoretically separated protein isomers with apparent molecular masses of 13, 15, and 16 kDa correspond to variants of the LECT1 protein-exhibiting peptide sequence polymorphisms that concern critical positions of the carbohydrate recognition domain (13 kDa: Leu51, Asn55, His130, Gly137; 15 kDa: Ser51, Asn55, Asn130, Gly137; 16 kDa: Ser51, Tyr55, Asn130, Glu137). Four residues, highly conserved in the galectin family, are substituted. None of the residues claimed to be involved in interactions with GalNAc alpha 1-3 moieties at an extended binding subsite of galectin-3 was identified in the corresponding positions of GCG. Apparently, the substitutions do not confer distinct binding characteristics to the GCG variants as evidenced by binding studies with a recombinantly expressed 15-kDa isoform. The natural isoforms as well as the recombinant 15-kDa isoform oligomerize by the formation of non-covalent heteromeric or homomeric complexes. A phosphorylation of the galectin was confirmed neither by mass spectrometry nor by alkaline phosphatase treatment combined with isoelectric focusing
Relationships of annelid subtaxa are controversially discussed and additional markers are necessarily needed to get further insights into their evolution. Due to their high content of information, mitochondrial genomes have been proven very useful in phylogenetic analyses. Whereas many complete mitochondrial genomes of arthropods are available, lophotrochozoan taxa are only scarcely represented and this is especially true for annelids. Here we present the complete mitochondrial genome of the orbiniid polychaete Orbinia latreillii. The circular genome is 15,558 bp in size and contains the same 37 genes as found in most other metazoans. As in the case for all studied annelids all genes are transcribed from the same strand. Compared with the known data from other annelids at least five gene translocations must be hypothesized for O. latreillii. A comparison of the available data shows that gene translocations within Annelida seem to be less frequent than in molluscs, but more frequent as previously assumed. Phylogenetic analyses of mitochondrial DNA sequence data and amino acid data support an inclusion Of Sipuncula within Annelida and a closer relationship to orbiniids is recovered for this taxon.
The communicatory significance of localised defecation sites in bushbuck (Tragelaphus scriptus)
(2006)
Like several other mammals, bushbuck (Tragelaphus scriptus) deposit faeces in specific localised defecation sites (LDS). A previous study has ruled out a function of LDS in the context of parasite avoidance. In this study, we investigated the communicatory significance of LDS. In a free ranging population, we tested whether LDS serve to demarcate home ranges, and/or if LDS are used for communication in a non-territorial context. In both sexes, LDS increased significantly in number towards the periphery of individual home ranges. However, the distribution pattern of LDS, as revealed by a nearest-neighbour mapping technique (nearest distances between LDS), did not support the idea that LDS serve home range/territory-demarcation because LDS did not follow a pattern of minimal nearest-neighbour distances along the margins of home ranges. We found females to urinate more often in LDS than males. Notably, information transfer was most frequent between sending (urinating or defecating) females and receiving/responding males (urination or defecation at places where a female had previously signalled). Our results suggest that LDS mainly serve for inter- sexual communication in bushbuck
The AtNFXL1 gene encodes a NF-X1 type zinc finger protein required for growth under salt stress
(2006)
The human NF-X1 protein and homologous proteins in eukaryotes represent a class of transcription factors which are characterised. by NF-X1 type zinc finger motifs. The Arabidopsis genome encodes two NF-X1 homologs, which we termed AtNFXL1 and AtNFXL2. Growth and survival was impaired in atnfxl1 knock-out mutants and AtNFXL1-antisense plants under salt stress in comparison to wild-type plants. In contrast, 35S: :AtNFXL1 plants showed higher survival rates. The AtNFXL2 protein potentially plays an antagonistic role. The Arabidopsis NF-X1 type zinc finger proteins likely are part of regulatory mechanisms, which protect major processes such as photosynthesis.
1. The in situ abundance, biomass and mean cell volume of Actinophrys sol (Sarcodina: Heliozoa), the top predator in an extremely acidic German mining lake (Lake 111; pH 2.65), were determined over three consecutive years (spring to autumn, 2001-03). 2. Actinophrys sol exhibited pronounced temporal and vertical patterns in abundance, biomass and mean cell volume. Increasing from very low spring densities, maxima in abundance and biomass were observed in late June/early July and September. The highest mean abundance recorded during the study was 7 x 10(3) Heliozoa L-1. Heliozoan abundance and biomass were higher in the epilimnion than in the hypolimnion. Actinophrys sol cells from this acidic lake were smaller than individuals of the same species found in other aquatic systems. 3. We determined the growth rate of A. sol using all potential prey items available in, and isolated and cultured from, Lake 111. Prey items included: single-celled and filamentous bacteria of unknown taxonomic affinity, the mixotrophic flagellates Chlamydomonas acidophila and Ochromonas sp., the ciliate Oxytricha sp. and the rotifers Elosa worallii and Cephalodella hoodi. Actinophrys sol fed over a wide-size spectrum from bacteria to metazoans. Positive growth was not supported by all naturally available prey. Actinophrys sol neither increased in cell number (k) nor biomass (k(b)) when starved, with low concentrations of single-celled bacteria or with the alga Ochromonas sp. Positive growth was achieved with single- celled bacteria (k = 0.22 +/- 0.02 d(-1); k(b) = -0.06 +/- 0.02 d(-1)) and filamentous bacteria (k = 0.52 +/- < 0.01 d(- 1); k(b) = 0.66 d(-1)) at concentrations greater than observed in situ, and the alga C. acidophila (up to k = 0.43 +/- 0.03 d(-1); k(b) = 0.44 +/- 0.04 d(-1)), the ciliate Oxytricha sp. (k = 0.34 +/- 0.01 d(-1)) and in mixed cultures containing rotifers and C. acidophila (k = 0.23 +/- 0.02-0.32 +/- 0.02 d(-1); maximum k(b) = 0.42 +/- 0.05 d(-1)). The individual- and biomass-based growth of A. sol was highest when filamentous bacteria were provided. 4. Existing quantitative carbon flux models for the Lake 111 food web can be updated in light of our results. Actinophrys sol are omnivorous predators supported by a mixed diet of filamentous bacteria and C. acidophila in the epilimnion. Heliozoa are important components in the planktonic food webs of 'extreme' environments
Chlamydomonas acidophila, a unicellular green alga, is a dominant phytoplankton species in acidic water bodies, facing severe environmental conditions such as low pH and high heavy metal concentrations. We examined the pH-, and temperature-dependent accumulation of heat-shock proteins in this alga to determine whether heat-shock proteins play a role in adaptation to their environment. Our results show increased heat-shock proteins accumulation at suboptimal pHs, which were not connected with any change in intracellular pH. In comparison to the mesophilic Chlamydomonas reinhardtii, the acidophilic species exhibited significantly higher accumulations of heat-shock proteins under control conditions, indicating an environmental adaptation of increased basal levels of heat-shock proteins. The results suggest that heat- shock proteins might play a role in the adaptation of C. acidophila, and possibly other acidophilic algae, to their extreme environment
A 13.94-m-long sediment core, collected from a medium-sized lake in the Qilian Mountains (NE Tibetan Plateau, China), was analysed palynologically at 81 horizons. The interpretation of indicator taxa yielded various vertical shifts of the vegetation belts. These palaeovegetation results have been checked with lake surface pollen spectra from 8 takes representing different altitudinal vegetation belts. Our main findings are the following: A short period of the late Marine Isotope Stage 3 (around similar to 46,000 yr ago) was characterized by interglacial temperature conditions with a tree line above its present-day altitude. During the LGM, the vicinity of the lake was not covered by ice but by sparse alpine vegetation and alpine deserts, indicating that the climate was colder by similar to 4-7 degrees C than today Markedly higher temperatures were inferred from higher arboreal pollen frequencies between similar to 13,000 and similar to 7000 yr ago with a Holocene temperature optimum and a maximal Picea-Betula mixed-forest expansion between similar to 9000 and similar to 7000 yr ago, when temperatures exceeded the present-day conditions by at least 1-2 degrees C. Alpine steppes and meadows and sub-alpine shrub vegetation dominated around the lake since the middle Holocene, suggesting that vegetation and climate conditions were exceptionally stable in comparison to previous periods.
This contribution describes a generator of stochastic time series of daily precipitation for the interior of Israel from c. 90 to 900 mm mean annual precipitation (MAP) as a tool for studies of daily rain variability. The probability of rainfall on a given day of the year is described by a regular Gaussian peak curve function. The amount of rain is drawn randomly from an exponential distribution whose mean is the daily mean rain amount (averaged across years for each day of the year) described by a flattened Gaussian peak curve. Parameters for the curves have been calculated from monthly aggregated, long-term rain records from seven meteorological stations. Parameters for arbitrary points on the MAP gradient are calculated from a regression equation with MAP as the only independent variable. The simple structure of the generator allows it to produce time series with daily rain patterns that are projected under climate change scenarios and simultaneously control MAP. Increasing within-year variability of daily precipitation amounts also increases among-year variability of MAP as predicted by global circulation models. Thus, the time series incorporate important characteristics for climate change research and represent a flexible tool for simulations of daily vegetation or surface hydrology dynamics.
This contribution describes a generator of stochastic time series of daily precipitation for the interior of Israel from c. 90 to 900 mm mean annual precipitation (MAP) as a tool for studies of daily rain variability. The probability of rainfall on a given day of the year is described by a regular Gaussian peak curve function. The amount of rain is drawn randomly from an exponential distribution whose mean is the daily mean rain amount (averaged across years for each day of the year) described by a flattened Gaussian peak curve. Parameters for the curves have been calculated from monthly aggregated, long-term rain records from seven meteorological stations. Parameters for arbitrary points on the MAP gradient are calculated from a regression equation with MAP as the only independent variable. The simple structure of the generator allows it to produce time series with daily rain patterns that are projected under climate change scenarios and simultaneously control MAP. Increasing within-year variability of daily precipitation amounts also increases among-year variability of MAP as predicted by global circulation models. Thus, the time series incorporate important characteristics for climate change research and represent a flexible tool for simulations of daily vegetation or surface hydrology dynamics.
Daylength is one of several parameters controlling flowering time in many plant species. The day length is perceived in leaves, but how the floral signal is transduced to the shoot apex via the phloem to induce flowering remains to be elucidated. This study aimed at the identification of new candidates involved in the induction of flowering by employing three plant species, Arabidopsis thaliana, Sinapis alba and Brassica napus in combination with transcript profiling by Affymetrix chip hybridization, metabolite profiling by gas chromatography – mass spectrometry and targeted protein analysis using antibodies. All analyses were performed on tissue-specific samples and focused on phloem sap or phloem exudates. To find common transcript and metabolite candidates potentially associated with the floral transition, two independent induction systems in Arabidopsis were used: a photoextension system, whereby plants received fourteen additional hours of light, and a parallel dexamethasone-inducible system, which was centered on the induction of the known flowering gene CONSTANS (CO). Identification of signals preceding the CO cascade was possible using the light extension regime, while downstream events dependent on CO activation were compared in both systems. Altogether, a number of interesting transcript and metabolite candidates were identified in both systems with some degree of overlap. Sinapis alba was used to investigate the universality of the floral signals between species. Comparisons of metabolite data revealed a few common candidates that may prove interesting for further studies. In addition, a targeted approach was carried out to investigate the presence of the Flowering Locus T (FT) protein during different stages of flower development using an antibody. Interesting changes in the sizes of antigens from rape phloem were seen and appeared consistent in Arabidopsis and to a lesser extent in Sinapis. Overall, the broad surveying approaches for transcripts and metabolites used in this study revealed several new potential candidates involved in the induction and/or regulation of flowering. As far as the protein work, additional experiments will reveal the link between FT and floral induction as well as its role in maintaining the floral state using the abovementioned plant species.
Soil seed banks near rubbing trees indicate dispersal of plant species into forests by wild boar
(2006)
Current knowledge about processes that generate long-distance dispersal of plants is still limited despite its importance for persistence of populations and colonization of new potential habitats. Today wild Large mammals are presumed to be important vectors for long-distance transport of diaspores within and between European temperate forest patches, and in particular wild boars recently came into focus. Here we use a specific habit of wild boar, i.e. wallowing in mud and subsequent rubbing against trees, to evaluate epizoochorous dispersal of vascular plant diaspores. We present soil seed bank data from 27 rubbing trees versus 27 control trees from seven forest areas in Germany. The mean number of viable seeds and the plant species number were higher in soil samples near rubbing trees compared with control trees. Ten of the 20 most frequent species were more frequent, and many species exclusively appeared in the soil samples near rubbing trees. The large number of plant species and seeds - more than 1000 per tree - in the soils near rubbing trees is difficult to explain unless the majority were dispersed by wild boar. Hooked and bristly diaspores, i.e. those adapted to epizoochory, were more frequent; however, many species with unspecialized diaspores occurred exclusively near rubbing trees. As opposed to plant species closely tied to forests species which occur both in forest and open vegetation and non-forest species were more frequent near rubbing trees compared with controls. These findings are consistent with previous studies on diaspore loads in the coats and hooves of shot wild boars. However, our method allows to identify the transport of diaspores from the open landscape into forest stands, where they might especially emerge after disturbance, and a clustered distribution of epizoochorically dispersed seeds. Moreover, accumulation of seeds of wetness indicators near rubbing trees demonstrates directed dispersal of plant species inhabiting wet places among remote wallows.
Soil seed banks near rubbing trees indicate dispersal of plant species into forests by wild boar
(2006)
Current knowledge about processes that generate long-distance dispersal of plants is still limited despite its importance for persistence of populations and colonization of new potential habitats. Today wild large mammals are presumed to be important vectors for long-distance transport of diaspores within and between European temperate forest patches, and in particular wild boars recently came into focus. Here we use a specific habit of wild boar, i.e. wallowing in mud and subsequent rubbing against trees, to evaluate epizoic dispersal of vascular plant diaspores. We present soil seed bank data from 27 rubbing trees versus 27 control trees from seven forest areas in Germany. The mean number of viable seeds and the plant species number were higher in soil samples near rubbing trees compared with control trees. Ten of the 20 most frequent species were more frequent, and many species exclusively appeared in the soil samples near rubbing trees. The large number of plant species and seeds – approximated > 1000 per tree – in the soils near rubbing trees is difficult to explain unless the majority were dispersed by wild boar. Hooked and bristly diaspores, i.e. those adapted to epizoochory, were more frequent, above that many species with unspecialised diaspores occurred exclusively near rubbing trees. Different to plant species closely tied to forest species which occur both in forest and open vegetation, and non-forest species were more frequent near rubbing trees compared with controls. These findings are consistent with previous studies on diaspore loads in the coats and hooves of shot wild boars. However, our method allows to identify the transport of diaspores from the open landscape into forest stands where they might especially emerge after disturbance, and a clustered distribution of epizoochorically dispersed seeds. Moreover, accumulation of seeds of wetness indicators near rubbing trees demonstrates directed dispersal of plant species inhabiting wet places between remote wallows.
Maldanids are usually divided into several subfamilies: Euclymeninae, Lumbriclymeninae, Maldaninae, Nicomachinae, Rhodininae, Clymenurinae, Notoproctinae, and Boguinae. The taxonomy of maldanids and the delimination of these taxa are mainly based on head morphology, total number of segments, chaetal structure, shape of the pygidium, and position of the anus. The maldanid ingroup relationships, as well as the monophyly of the proposed subfamilies, have so far not been investigated. Pilgrim (1977) described a shift of the notopodial chaetal rows from a transverse direction in anterior chaetigers to a more longitudinal one in posterior chaetigers in Clymene torquata (Leidy, 1855) and Euclymene oerstedii (Claparede, 1863), both belonging to the Euclymeninae. We investigated several maldanid species to assess the usefulness of this character for maldanid systematics and used 3D-reconstruction techniques to reveal the complete geometry of the chaetal sacs. Our investigation shows that a shift is apparent in Euclymene, Axiothella, Johnstonia (all Euclymeninae) and Clymenura (Clymenurinae), but absent in species like Maldane sarsi (Malmgren, 1865), Metasychis disparidentata (Moore, 1904) (both Maldaninae) and Petaloproctus borealis Ardwisson, 1906 (Nicomachinae). The shift is not typical for sedentary polychaetes and is apomorphic within maldanid polychaetes. It thus argues for a close relationship between Euclymeninae and Clymenurinae. The investigation of further maldanid species of different subfamilies may shed additional light on maldanid systematics.
Flüssigkeitssekretion und Proteinsekretion werden in Speicheldrüsen von Insekten über Hormone und Neurotransmitter gesteuert. Diese entfalten ihre physiologische Wirkung in den sekretorischen Drüsenzellen hauptsächlich über den zyklischen Adenosinmonophosphat (cAMP)-Signalweg und den Inositoltrisphosphat (IP<SUB>3</SUB>) / Ca<sup>2+-Signalweg. Die Mechanismen möglicher Wechselwirkungen zwischen diesen Signalwegen und ihre physiologischen Auswirkungen sind unzureichend bekannt. Im Mittelpunkt dieser Arbeit stand die Frage, ob und wie sich der Ca<sup>2+-Signalweg und der cAMP-Signalweg in der Speicheldrüse der Diptere Calliphora vicina beeinflussen. Substanzen wie 5-Fluoro-α-Methyltryptamin und Histamin wurden in früheren Arbei-ten als Agonisten genutzt, um in den Speicheldrüsen von C. vicina selektiv den cAMP-Signalweg (getrennt vom IP<SUB>3</SUB>/Ca<sup>2+-Signalweg) zu aktivieren. Es zeigte sich in transepithelialen Potentialmessungen und mikrofluorometrischen Ca<sup>2+-Untersuchungen, dass beide Substanzen sowohl den cAMP-Weg als auch den Ca<sup>2+-Signalweg aktivierten. Die physiologischen Ursachen der Histamin-induzierten Ca<sup>2+-Erhöhung wurden genauer untersucht. Zusammengefasst zeigten diese Untersuchungen, dass Histamin wie 5-HT den cAMP-Weg und die Phosphoinositidkaskade aktivierte. Im Gegensatz zu den 5-HT-induzierten Ca<sup>2+-Oszillationen, welche durch interzelluläre Ca<sup>2+-Wellen synchronisiert werden, verursachte Histamin bei niedrigen Konzentrationen lokale Ca<sup>2+-Oszillationen in einzelnen Zellen (keine Wellen). Bei höheren Histamin-Konzentrationen war eine anhaltende Ca<sup>2+-Erhöhung oder ein synchrones <quote>Ca<sup>2+-beating</quote> in der gesamten Drüse zu beobachten. Des Weiteren wurde die Frage untersucht, ob eine Erhöhung der intrazellulären cAMP-Konzentration den IP<SUB>3</SUB> Ca<sup>2+-Signalweg in den Epithelzellen der Speicheldrüse beeinflussen kann. Es zeigte sich, dass cAMP den durch schwellennahe 5-HT-Konzentrationen induzierten Ca<sup>2+-Anstieg verstärkte. Diese Verstärkung wurde durch eine PKA-vermittelte Sensitivierung des IP<SUB>3</SUB>-Rezeptor/Ca<sup>2+-Kanals für IP<SUB>3</SUB> verursacht. Immunzytochemische Untersuchungen deuten dar-auf hin, dass die Proteinkinase A eng mit dem IP<SUB>3</SUB>-Rezeptor/Ca<sup>2+-Kanal assoziiert ist. Diese Messungen zeigen erstmals, dass auch bei Invertebraten der Botenstoff cAMP, PKA-vermittelt, den IP<SUB>3</SUB>-Rezeptor/Ca<sup>2+-Kanal des ER für IP<SUB>3</SUB> sensitiviert.
In semi-arid savannas, unsustainable land use can lead to degradation of entire landscapes, e.g. in the form of shrub encroachment. This leads to habitat loss and is assumed to reduce species diversity. In BIOTA phase 1, we investigated the effects of land use on population dynamics on farm scale. In phase 2 we scale up to consider the whole regional landscape consisting of a diverse mosaic of farms with different historic and present land use intensities. This mosaic creates a heterogeneous, dynamic pattern of structural diversity at a large spatial scale. Understanding how the region-wide dynamic land use pattern affects the abundance of animal and plant species requires the integration of processes on large as well as on small spatial scales. In our multidisciplinary approach, we integrate information from remote sensing, genetic and ecological field studies as well as small scale process models in a dynamic region-wide simulation tool. <hr> Interdisziplinäres Zentrum für Musterdynamik und Angewandte Fernerkundung Workshop vom 9. - 10. Februar 2006.
Rhodobacter capsulatus xanthine dehydrogenase (XDH) is a cytoplasmic enzyme with an (alpha beta) 2 heterodimeric structure that is highly identical to homodimeric eukaryotic xanthine oxidoreductases. The crystal structure revealed that the molybdenum cofactor (Moco) is deeply buried within the protein. A protein involved in Moco insertion and XDH maturation has been identified, which was designated XdhC. XdhC was shown to be essential for the production of active XDH but is not a subunit of the purified enzyme. Here we describe the purification of XdhC and the detailed characterization of its role for XDH maturation. We could show that XdhC binds Moco in stoichiometric amounts, which subsequently can be inserted into Moco-free apo-XDH. A specific interaction between XdhC and XdhB was identified. We show that XdhC is required for the stabilization of the sulfurated form of Moco present in enzymes of the xanthine oxidase family. Our findings imply that enzyme-specific proteins exist for the biogenesis of molybdoenzymes, coordinating Moco binding and insertion into their respective target proteins. So far, the requirement of such proteins for molybdoenzyme maturation has been described only for prokaryotes
We examined chemical communication in male and female European pond turtles (Emys orbicularis). In simultaneous binary choice tests, a focal animal was given a choice between pheromones from a conspecific and a choice chamber containing untreated water. Females did not show a preference, both when male and when female stimuli were presented. On the contrary, males preferred the odor of a female over untreated water, suggesting that males actively search for females. The strength of preference was positively correlated with the body size difference between the female and the focal male, indicating that males prefer to mate with larger females. Female fecundity is positively correlated with female size in E. orbicularis, which may account for male choosiness. No overall preference for the stimulus animal was observed when males were presented cues from another male. However, the strength of preference was negatively correlated with the difference in body size. Males avoided large males, but oriented towards smaller stimulus males. This reflects that males form dominance hierarchies, where large males aggressively attack smaller ones. Far-range chemical communication probably enables males to minimize the risk of costly aggressive interactions. This is, to our knowledge, the first study on the role of chemical cues for inter-and intrasexual communication in the European pond turtle
More than one century ago, sturgeons were prevalent species in the fish communities of all major German rivers both in the North and the Baltic seas drainages. Since then, the populations declined rapidly due to river damming, overfishing and pollution. The last sturgeon catches in the Baltic drainage system occurred during the late 1960ies. Only a few individual captures have been reported during the last 30 years with the most recent records in the Lake Ladoga ( Russia), where the last confirmed catch was recorded in 1984, and a single individual caught off Estonia in 1996. Today, sturgeons are considered missing or extinct in German waters. First attempts for remediation of the species were undertaken in the mid 1990ies. Subsequently, phylogenetic and population genetic analyses of the species were carried out using mtDNA, microsatellites, and nuclear markers ( SNPs). These genetic analyses using recent and historic material have proven the existence of two different species in the Baltic Sea in what was previously considered to represent the European Atlantic sturgeon only. In the Baltic Sea, the American Atlantic sturgeon ( A. oxyrinchus) succeeded to colonize this brackish water system during the Middle Ages. In the North Sea, the European Atlantic sturgeon ( A. sturio) is considered to be the endemic species. These results led to the separation of the remediation activities in the North Sea and the Baltic Sea tributaries. Further studies on the mechanism that lead to the extinction of A. sturio in Germany and the subsequent succession of the A. oxyrinchus mtDNA haplotype are currently been carried out. Broodstock development using the northernmost populations of A. oxyrinchus is currently under way. As a further prerequisite to re-introduce this species into the Baltic, the evaluation of the status of critical habitats for the early life stages of the American Atlantic sturgeon in the River Odra has been performed in collaboration with the Institute for Inland Fisheries of Poland. Alternative fisheries techniques, based on the data of by-catch of exotic sturgeons in the fishery, are presently developed in close cooperation with the fishery to reduce fisheries related mortality in juvenile sturgeons upon release. Monitoring of habitat utilization and migration characteristics of juvenile fish upon experimental release will have to be carried out shortly, using acoustic telemetry, with the aim to follow the fate of the released fish and to determine the best time-size-release-window for future release programmes.
Context: Placental and circulatory soluble fms-like tyrosine kinase 1 (sFlt1) has proven to be elevated in pregnant women with preeclampsia, a disease characterized by hypertension, proteinuria, and endothelial dysfunction. Recent studies also demonstrated an autoantibody against the angiotensin II type 1 (AT1) receptor (AT1-AA) in that disease. Objective: Both factors are discussed as key players in the etiology of preeclampsia. However, it has not yet been clarified whether these two circulating factors correlate and whether synergy determines the severity of pathology. Design: AT1-AA was retrospectively determined by a bioassay and sFlt1 by an ELISA. Patients: Serum from second-trimester pregnancies with normal or abnormal uterine perfusion and in women at term with or without pregnancy pathology was analyzed. Results: Most of the preeclamptic patients were characterized by high sFlt1 levels and the presence of AT1-AA, although the agonistic effects of the antibody did not correlate with the sFlt1 concentrations (P = 0.85). Although AT1- AA was also detected in second-trimester pregnancies evidencing abnormal uterine perfusion without later pathology, sFlt1 was not significantly elevated in these pregnancies, compared with those with normal uterine perfusion. However, whereas women with abnormal perfusion and later pregnancy pathology did not differ in AT1-AA, compared with those with normal outcome, sFlt1 was significantly increased. Again, the two factors did not correlate (P = 0.15). Conclusions: We conclude that AT1-AA bioactivity and sFlt1 concentrations do not correlate, are not mutually dependent, and are thus probably involved in distinct pathogenetic mechanisms. Both factors in combination may not be causative for the early impaired trophoblast invasion and pathological uterine perfusion
Cave animals are widely recognised as model organisms to study regressive evolutionary processes like the reduction of eyes. In this paper, we report on the regressive evolution of species discrimination in the cave molly, Poecilia mexicana, which, unlike other cave fishes, still has functional eyes. This allowed us to examine the response to both visual and non-visual cues involved in species discrimination. When surface-dwelling females were given a chance to associate with either a conspecific or a swordtail (Xiphophorus hellerii) female, they strongly preferred the conspecific female both when multiple cues and when solely visual cues were available to the female. No association preference was observed when only non-visual cues were provided. In contrast, cave-dwelling females showed no preference under all testing conditions, suggesting that species recognition mechanisms have been reduced. We discuss the role of species discrimination in relation to habitat differences.
A novel method for the encapsulation of biomacromolecules, such as nucleic acids and proteins, into polyelectrolyte microcapsules is described. Fluorescence-labelled double-stranded DNA and human serum albumin (HSA) are used as model substances for encapsulation in hollow microcapsules templated on human erythrocytes. The encapsulation procedure involves an intermediate drying C, step. The accumulation of DNA and HSA in the capsules is observed by confocal laser scanning microscopy, UV spectroscopy, and flourimetry. The mechanism of encapsulation is discussed
The advent of large-scale and high-throughput technologies has recently caused a shift in focus in contemporary biology from decades of reductionism towards a more systemic view. Alongside the availability of genome sequences the exploration of organisms utilizing such approach should give rise to a more comprehensive understanding of complex systems. Domestication and intensive breeding of crop plants has led to a parallel narrowing of their genetic basis. The potential to improve crops by conventional breeding using elite cultivars is therefore rather limited and molecular technologies, such as marker assisted selection (MAS) are currently being exploited to re-introduce allelic variance from wild species. Molecular breeding strategies have mostly focused on the introduction of yield or resistance related traits to date. However given that medical research has highlighted the importance of crop compositional quality in the human diet this research field is rapidly becoming more important. Chemical composition of biological tissues can be efficiently assessed by metabolite profiling techniques, which allow the multivariate detection of metabolites of a given biological sample. Here, a GC/MS metabolite profiling approach has been applied to investigate natural variation of tomatoes with respect to the chemical composition of their fruits. The establishment of a mass spectral and retention index (MSRI) library was a prerequisite for this work in order to establish a framework for the identification of metabolites from a complex mixture. As mass spectral and retention index information is highly important for the metabolomics community this library was made publicly available. Metabolite profiling of tomato wild species revealed large differences in the chemical composition, especially of amino and organic acids, as well as on the sugar composition and secondary metabolites. Intriguingly, the analysis of a set of S. pennellii introgression lines (IL) identified 889 quantitative trait loci of compositional quality and 326 yield-associated traits. These traits are characterized by increases/decreases not only of single metabolites but also of entire metabolic pathways, thus highlighting the potential of this approach in uncovering novel aspects of metabolic regulation. Finally the biosynthetic pathway of the phenylalanine-derived fruit volatiles phenylethanol and phenylacetaldehyde was elucidated via a combination of metabolic profiling of natural variation, stable isotope tracer experiments and reverse genetic experimentation.
Profiling of plant secondary metabolites is still a very difficult task. Liquid chromatography (LC) or capillary electrophoresis hyphenated with different kinds of detectors are methods of choice for analysis of polar, thermo labile compounds with high molecular masses. We demonstrate the applicability of LC combined with UV diode array or/and mass spectrometric detectors for the unambiguous identification and quantification of flavonoid conjugates isolated from Arahidopsis thaliana leaves of different genotypes and grown in different environmental conditions. During LC/UV/MS/MS analyses we were able to identify tetra-, tri, and di-glycosides of kaempferol, quercetin and isorhamnetin. Based on our results we can conclude that due to the co-elution of different chemical compounds in reversed phase H PLC systems the application of UV detectors does not allow to precisely profile all flavonoid conjugates existing in A. thaliana genotypes. Using MS detection it was possible to unambiguously recognize the glycosylation patterns of the aglycones. However, from the mass spectra we could not conclude neither the anomeric form of the C-1 carbon atoms of sugar moieties in glycosidic bonds between sugars or sugar and aglycone nor the position of the second carbon involved in disaccharides. The applicability of collision induced dissociation techniques (CID MS/MS) for structural analyses of the studied group of plant secondary metabolites with two types of analyzers (triple quadrupole or ion trap) was demonstrated.
Predicting the breeding success of large raptors in arid southern Africa : a first assessment
(2006)
Raptors are often priorities for conservation efforts and breeding success is a target measure for assessing their conservation status. The breeding success of large raptors in and southern Africa is thought to be higher in years of high rainfall. While this correlation has been found in several studies, it has not yet been shown for data from a wider geographical area. In conservation research, it is important to explore the differences between spatially- separated populations to estimate and to compare their conservation status, and to deduce specific management strategies. Using a theoretical approach, we develop a simplistic model to explain the breeding success-rainfall relationship in large African raptors at larger spatial scales. Secondly, we validate this model and we show that the inclusion of field data leads to consistent predictions. In particular, we recommend that the average size of the 'effective territory' should be included in the relationship between annual rainfall and breeding success of raptors in and southern Africa. Accordingly, we suggest that breeding success is a function of precipitation and inter- nest distance. We present a new measure of territory quality depending on rainfall and territory size. We suggest that our model provides a useful first approach to assess breeding success in large raptors of and southern Africa. However, we strongly emphasise the need to gather more data to further verify our model. A general problem in conservation research is to compare the status of populations assessed in different study areas under changing environmental conditions. Our simplistic approach indicates that this problem can be overcome by using a weighted evaluation of a target measure (i.e. breeding success), taking regional differences into account
Flagellates are important bacterial grazers in most planktonic food webs. The prey-size preference of the mixotrophic flagellate, Ochromonas sp. (Chrysophyceae), isolated from an extremely acidic lake, Lake 111 (pH 2.6), was determined using fluorescently labelled microspheres (beads). According to grazing experiments with cultured bacteria, also isolated from Lake 111, the potential grazing impact on Lake 111"s single-celled bacterial production was calculated. Ochromonas sp. ingested the smallest beads offered (0.5 µm diameter) at the highest rate. Ingestion rate declined with increasing bead size. The highest prey volume-specific ingestion was measured for Ochromonas sp. feeding on intermediate-sized beads (1.9 µm). Ingestion rates were low due in part to the large fraction of inactive flagellates observed. According to the bacterial ingestion rate, a mean of 88% (epilimnion) and 68% (hypolimnion) of in situ single- celled bacterial production is potentially grazed daily by Ochromonas sp. In the epilimnion of Lake 111, the heterotrophic carbon gain is three times higher than the autotrophic production. Alongside carbon uptake, Ochromonas sp. also benefits from ingesting bacteria through the uptake of phosphorus. A biovolume minimum corresponding to the prey size at which Ochromonas sp. feeds most efficiently occurred in the Lake 111 epilimnetic bacterial community, implying top-down control of the bacterial community by Ochromonas sp.
The pollination biology of three mass flowering Utricularia species of the Indian Western Ghats, U. albocaerulea, U. purpurascens, and U. reticulata, was studied for the first time by extensive observation of flower visitors, pollination experiments, and nectar analyses. The ephemerality of the Utricularia habitats on lateritic plateaus, weather conditions adverse to insects, lack of observations of flower visitors to other Utricularia spp., and the predominance of at least. facultative autogamy in the few Utricularia species studied so far suggested that an autogamous breeding system is the common case in the genus. In contrast, we showed that the studied populations are incapable of autonomous selfing, or that it is an event of negligible rarity, although P/O was similarily low as in autogamous species investigated by other authors. In all three species the spatial arrangement of the reproductive organs makes an insect vector necessary for pollen transfer between and within flowers. However, U. purpurascens and U. reticulata are highly self-compatible, which allows for visitor-mediated auto-selfing and geitonogamy on inflorescence and clone level. Floral nectar is present in extremely small volumes in all three species, but sugar concentrations are high. More than 50 species of bees, butterflies, moths, hawk moths, and clipterans were observed to visit the flowers, and flower morphology facilitated pollination by all observed visitors. The results are discussed in the context of the phenological characteristics of the studied species, especially the phenomenon of mass flowering, and the environmental conditions of their habitats.
Plastidial phosphorylase is required for normal starch synthesis in Chlamydomonas reinhardtii
(2006)
Among the three distinct starch phosphorylase activities detected in Chlamydomonas reinhardtii, two distinct plastidial enzymes (PhoA and PhoB) are documented while a single extraplastidial form (PhoC) displays a higher affinity for glycogen as in vascular plants. The two plastidial phosphorylases are shown to function as homodimers containing two 91-kDa (PhoA) subunits and two 110-kDa (PhoB) subunits. Both lack the typical 80-amino-acid insertion found in the higher plant plastidial forms. PhoB is exquisitely sensitive to inhibition by ADP-glucose and has a low affinity for malto-oligosaccharides. PhoA is more similar to the higher plant plastidial phosphorylases: it is moderately sensitive to ADP-glucose inhibition and has a high affinity for unbranched malto-oligosaccharides. Molecular analysis establishes that STA4 encodes PhoB. Chlamydomonas reinhardtii strains carrying mutations at the STA4 locus display a significant decrease in amounts of starch during storage that correlates with the accumulation of abnormally shaped granules containing a modified amylopectin structure and a high amylose content. The wild-type phenotype could be rescued by reintroduction of the cloned wild-type genomic DNA, thereby demonstrating the involvement of phosphorylase in storage starch synthesis.
Peridinin-chlorophyll a-protein (PCP) is a unique antenna complex in dinoflagellates that employs peridinin (a carotenoid) as its main light-harvesting pigment. Strong excitonic interactions between peridinins, as well as between peridinins and chlorophylls (Chls) a, can be expected from the short intermolecular distances revealed by the crystal structure. Different experimental approaches of nonlinear polarization spectroscopy in the frequency domain (NLPF) were used to investigate the various interactions between pigments in PCP of Amphidinium carterae at room temperature. Lineshapes of NLPF spectra indicate strong excitonic interactions between the peridinin's optically allowed S-2 (1Bu(+)) states. A comprehensive subband analysis of the distinct NLPF spectral substructure in the peridinin region allows us to assign peridinin subbands to the two Chls a in PCP having different S-1-state lifetimes. Peridinin subbands at 487, 501, and 535 nm were assigned to the longer-lived Chl, whereas a peridinin subband peaking at 515 nm was detected in both clusters. Certain peridinin(s), obviously corresponding to the subband centered at 487 nm, show(s) specific (possibly Coulombic?) interaction between the optically dark S-1(2A(g)(-)) and/or intramolecular charge- transfer (ICT) state and S-1 of Chl a. The NLPF spectrum, hence, indicates that this peridinin state is approximately isoenergetic or slightly above S-1 of Chl a. A global subband analysis of absorption and NLPF spectra reveals that the Chl a Q(y)-band consists of two subbands ( peaking at 669 and 675 nm and having different lifetimes), confirmed by NLPF spectra recorded at high pump intensities. At the highest applied pump intensities an additional band centered at <= 660 nm appears, suggesting-together with the above results-an assignment to a low-dipole moment S-0-> S-1/ICT transition of peridinin
The photosensitive microvilli of Drosophila photoreceptors R1-R6 are not aligned in parallel over the entire length of the visual cells. In the distal half of each cell, the microvilli are slightly tilted toward one side and, in the proximal half, extremely toward the opposite side. This phenomenon, termed rhabdomere twisting, has been known for several decades, but the developmental and cell biological basis of rhabdomere twisting has not been studied so far. We show that rhabdomere twisting is also manifested as molecular polarization of the visual cell, because phosphotyrosine- containing proteins are selectively partitioned to different sides of the rhabdomere stalk in the distal. and proximal sections of each R1-R6 photoreceptor. Both the asymmetrical segregation of phosphotyrosine proteins and the tilting of the microvilli occur shortly before eclosion of the flies, when eye development in all other aspects is considered to be essentially complete. Establishment of rhabdomere twisting occurs in a light-independent manner, because phosphotyrosine staining is unchanged in dark-reared wild-type flies and in mutants with defects in the phototransduction cascade, ninaE(17) and norpA(P24). We conclude that antiphosphotyrosine immunofluorescence can be used as a light microscopic probe for the analysis of rhabdomere twisting and that microvilli tilting represents a type of planar cell polarity that is established by an active process in the last phase of photoreceptor morphogenesis, just prior to eclosion of the flies.