Inhibition by mevinolin of plant growth, sterol formation and pigment accumulation

To obtain information on the importance of a functional mevalonate synthesis for plant growth and development, we investigated the effect of mevinolin, a highly specific inhibitor of 3-hydroxy-3-methylglutaryl (HMG) coenzyme A reductase (the mevalonate-producing enzyme) on growth, sterol accumulation and pigment formation of radish seedlings (Raphanus sativus L. cv. Saxa Treib) and in part also wheat seedlings (Triticum aestivum L. cv. Kolibri). Mevinolin applied during germination inhibits root elongation and development of lateral roots in etiolated and light-grown radish seedlings. This effect cannot be overcome by exogenous GA₃, but by addition of mevalonic acid, the product of the internally inhibited reaction. This emphazises the specifity of the mevinolin effect and indicates that the biosynthesis of mevalonic acid is a mandatory requirement for root growth. In light-grown radish seedlings mevinolin also affects hypocotyl length-growth and inhibits sterol accumulation, but has little effect on the chlorophyll and carotenoid accumulation in the chloroplasts of the cotyledons. This indicates the possible presence of an independent mevalonate synthesizing pathway within the plastids and suggests a low transport rate of mevinolin from the radish roots to the cotyledons. When mevinolin is directly applied to the leaves at higher concentrations, it also reduces the light-induced chlorophyll and carotenoid accumulation as has been shown with etiolated primary leaves of wheat. This inhibition is age-dependent and proceeds to a higher extent in older than in younger etiolated leaf tissue. From our results we conclude that plastids possess an independent HMG-CoA reductase. In the cotyledons of radish, mevinolin seems to induce a senescence retardation and sun-type growth response, as has been evaluated by measuring the fast and slow chlorophyll fluorescence induction kinetics (Kautsky effect). These responses may be due to inhibitor-induced changes in the intracellular phytohormone balance.     

Uptake of the Herbicide Diuron as Visualised by the Fluorescence Imaging Technique

A new fluorescence imaging system for monitoring the uptake of the PSII-herbicide diuron (OCMU) was tested in tobacco leaves. UV-laser-induced (Λ_exc = 355 nm) fluorescence images were collected for blue fluorescence F440 (Λ_em = 440 nm), green fluorescence F520 (Λ_em = 520 nm), red chlorophyll fluorescence F690 (Λ_em = 690 nm) and for far-red chlorophyll fluorescence F740 (Λ_em = 740 nm). Diuron-treated leaf parts exhibited a higher red and far-red chlorophyll fluorescence emission (F690 and F740) than untreated leaf halves, whereas the blue and green fluorescence, F440 and F520, remained unaffected. As a consequence, the fluorescence ratios blue/red (F440/F690) and blue/far-red (F440/F740) significantly decreased in diuron-treated leaf parts. The time course of diuron uptake into the leaf could be followed by fluorescence images taken 10 and 30 min after diuron application. The novel high resolution fluorescence imaging method supplies information on the herbicide uptake of each point of the leaf area. Its great advantage as compared to the point data fluorescence measurements applied so far is discussed.     

Biosynthesis of isoprenoids in higher plant chloroplasts proceeds via a mevalonate-independent pathway

Isopentenyl diphosphate (IPP) is the biological C₅ precursor of isoprenoids. By labeling experiments using [1-¹³C]glucose, higher plants were shown to possess two distinct biosynthetic routes for IPP biosynthesis: while the cytoplasmic sterols were formed via the acetate/mevalonate pathway, the chloroplast-bound isoprenoids (β-carotene, lutein, prenyl chains of chlorophylls and plastoquinone-9) were synthesized via a novel IPP biosynthesis pathway (glyceraldehyde phosphate/pyruvate pathway) which was first found in eubacteria and a green alga. The dichotomy in isoprenoid biosynthesis in higher plants allows a reasonable interpretation of previous odd and inconclusive results concerning the biosynthesis of chloroplast isoprenoids, which so far had mainly been interpreted in the frame of models using compartmentation of the mevalonate pathway.     

Die Plastoglobuli von Spinat,ihre Grö\e,Isolierung und Lipochinonzusammensetzung

Zusammenfassung Die Grö\e und HÄufigkeit der osmiophilen Plastoglobuli nimmt mit steigendem Alter der Spinatchloroplasten zu. Die Plastoglobuli besitzen in ganzen oder aufgebrochenen Chloroplasten, in Sedimenten und in isolierter Form die gleiche Grö\enverteilung wiein situ. Es gibt keine Hinweise, da\ beim Ultraschallaufschlu\ ganzer Chloroplasten Plastoglobuli durch Entmischung der Thylakoide entstehen. Beim Zentrifugieren von broken chloroplasts wird nur ein kleiner Teil der Plastoglobuli freigesetzt. Der grö\ere Teil befindet sich mit den Chloroplastenbruchstücken im Sediment.Wie bei anderen Pflanzen enthalten auch die Plastoglobuli von Spinat vorwiegend die lipophilen Plastidenchinone vom Benzochinontyp (Plastochinon 45, Plastohydrochinon,-Tocopherol und-Tocochinon), geringe Mengen an Vitamin K₁ und Spuren an Carotinoiden, jedoch keine Chlorophylle. Zwischen dem Lipochinongehalt der Chloroplasten und der Grö\e und HÄufigkeit der osmiophilen Plastoglobuli besteht eine direkte Korrelation.

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The plastoglobuli of spinach: Their size, isolation, and lipoquinone composition

Summary The size and frequency of the osmiophilic plastoglobuli of spinach chloroplasts increase with increasing age of chloroplasts. The plastoglobuli of whole or broken chloroplasts, of sediments and in the isolated form show the same size distribution asin situ. There is no evidence that plastoglobuli may arise from thylakoid lipids by ultrasonic treatment of whole chloroplasts. During the centrifugation of broken chloroplasts only a small portion of the plastoglobuli is releazed, the greater portion remains in the sediment together with the chloroplast fragments.The plastoglobuli of spinach—like those of other plants—mainly contain the excess plastidquinones of the benzoquinone type (plastoquinone 45, plastoquinol,-tocopherol,-tocoquinone), little vitamin K₁ (a naphthoquinone), traces of carotenoids, but no chlorophylls. The lipoquinone content of spinach chloroplasts increases parallel to the augmentation of the size and frequency of the plastoglobuli.

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Der Deutschen Forschungsgemeinschaft danke ich für ihre Unterstützung und FrÄuleinMargarethe Wirtz für tüchtige technische Mitarbeit.     

The novel sorting nexin SNX33 interferes with cellular PrP formation by modulation of PrP shedding

The cellular prion protein (PrP^c) is a glycosyl-phosphatidylinositol (GPI)-anchored protein trafficking in the secretory and endocytic pathway and localized mainly at the plasma membrane. Conversion of PrP^c into its pathogenic isoform PrP^Sc is associated with pathogenesis and transmission of prion diseases. Intramolecular cleavage in the middle, the extreme C-terminal part or within the GPI anchor and shedding of PrP^c modulate this conversion process by reducing the substrate for prion formation. These phenomena provide similarities with the processing of amyloid precursor protein in Alzheimer's disease. Sorting nexins are a family of proteins with important functions in protein trafficking. In this study, we investigated the role of the newly described sorting nexin 33 (SNX33) in trafficking and processing of PrP^c. We found that overexpression of SNX33 in neuronal and non-neuronal cell lines resulted in increased shedding of full-length PrP^c from the plasma membrane and modulated the rate of PrP^c endocytosis. This was paralleled by reduction of PrP^Sc formation in persistently and newly infected cells. Using deletion mutants, we demonstrate that production of PrP fragment N1 is not influenced by SNX33. Our data provide new insights into the cellular mechanisms of PrP^c shedding and show how this can affect cellular PrP^Sc conversion.     

Comparison of verdeluz orange G and modified Gallego stains

Tumors of the oral cavity include combinations of hard and soft tissues that may be difficult to identify using routine hematoxylin and eosin (H & E) staining. Although combination stains can demonstrate hard and soft tissues, trichrome stains, such as VanGieson and Masson, cannot differentiate dental hard tissues, such as dentin, cementum and osteoid. Modified Gallegos (MGS) and verdeluz orange G-acid fuchsin (VOF) stains can differentiate components of teeth. We used 10 tissue sections of decalcified bone and 10 pathologic tissue sections that contained different calcified tissues including peripheral ossifying fibroma, odontoma, central ossifying fibroma and cemento-ossifying fibroma. Sections were stained with H & E, VOF or MGS. H and E stained both hard tissues pink. VOF stained bone purple-red, cementum red and collagen blue. MGS stained bone green-blue, cementum red and collagen blue. VOF staining intensity and differentiation was better than MGS staining. VOF staining demonstrated hard tissue components distinctly and exhibited good contrast with the surrounding connective tissue. VOF also is a simple, single step, rapid staining procedure.     

Anatomically based lower limb nerve model for electrical stimulation

Background  

Functional Electrical Stimulation (FES) is a technique that aims to rehabilitate or restore functionality of skeletal muscles using external electrical stimulation. Despite the success achieved within the field of FES, there are still a number of questions that remain unanswered. One way of providing input to the answers is through the use of computational models.     

Die Hemmung der Carotinoidsynthese im Stickstoffmangel

Summary The ability to synthesize carotenoids during normal or strongly reduced nitrogen supply was tested in a higher plant (Hordeum vulgare L.) and in a green alga (Scenedesmus obliquus). Though carotenoids are nitrogenfree, their synthesis is strongly inhibited during nitrogen deficiency. The formation of secondary carotenoids, which apparently proceeds at the expense of the primary carotenoid components, occurs in Scenedesmus but not in Hordeum plants. It is concluded that the dimerisation of geranly-geranyl-pyrophosphate is the limiting step in the carotenoid synthesis during nitrogen deficiency. The synthesis of prenyl chains is less affected by nitrogen deficiency.