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Publications - Stress and Develop Biology

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Publications

Clemens, S.; Bloss, T.; Vess, C.; Neumann, D.; Nies, D. H.; zur Nieden, U.; A Transporter in the Endoplasmic Reticulum of Schizosaccharomyces pombe Cells Mediates Zinc Storage and Differentially Affects Transition Metal Tolerance J. Biol. Chem. 277, 18215-18221, (2002) DOI: 10.1074/jbc.M201031200

The cation diffusion facilitator (CDF) family represents a class of ubiquitous metal transporters. Inactivation of a CDF in Schizosaccharomyces pombe, Zhf, causes drastically different effects on the tolerance toward various metals. A deletion mutant is Zn2+/Co2+-hypersensitive yet displays significantly enhanced Cd2+ and Ni2+ tolerance. Accumulation of zinc, cobalt, and cadmium is reduced in mutant cells. Non-vacuolar zinc content, as measured by analytical electron microscopy, is lower in zhf− cells compared with wild-type cells in the presence of elevated Zn2+concentrations. The protective effect against cadmium toxicity is independent of the phytochelatin detoxification pathway. Phytochelatin synthase-deficient cells show extremely enhanced (about 200-fold) cadmium tolerance when zhf is disrupted. Immunogold labeling indicates endoplasmic reticulum (ER) localization of Zhf. Electron spectroscopic imaging shows that accumulation of zinc coincides with Zhf localization, demonstrating a major role of the ER for metal storage and the involvement of Zhf in cellular zinc homeostasis. Also, these observations indicate that Cd2+ions exert their toxic effects on cellular metabolism in the ER rather than in the cytosol.
Books and chapters

Bruns, I.; Sutter, K.; Neumann, D.; Krauss, G.-J.; Glutathione accumulation - a specific response of mosses to heavy metal stress 389-391, (2000)

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Publications

Wollgiehn, R.; Neumann, D.; Metal Stress Response and Tolerance of Cultured Cells from Silene vulgaris and Lycopersicon peruvianum: Role of Heat Stress Proteins J. Plant Physiol. 154, 547-553, (1999) DOI: 10.1016/S0176-1617(99)80296-X

The influence of the heavy metal ions Hg++, Cu++, Cd++ and Zn++ and of arsenite on growth, amino acid uptake, protein- and heat shock protein synthesis was investigated in cell cultures of a heavy metal tolerant Silene vulgaris and the sensitive Lycopersicon peruvianum.A distinct tolerance of Silene cell growth in comparison to tomato cells against Cu, Cd and Zn was observed. Synthesis of the small heat stress proteins was induced in both species, however, with quantitative differences depending on species and metal ion. While in tomato cells all metals induce HSP synthesis, in Silene the induction by Cu and Cd was found to be significantly lower in comparison to Hg and arsenite, and Zn did not induce HSP. Therefore, generally a lower tolerance of the cells against a metal is connected with a higher HSP synthesis. From comparison of cell growth and HSP accumulation in the presence of metal ions it was further concluded that HSP synthesis is a part of HM stress response of tolerant and non tolerant cells as under heat shock, but HSPs are not responsible for the heritable metal tolerance of Silene.In contrast to heat shock, metal stress does not inhibit the cell protein synthesis directly. In cultured tomato and Silene cells the inhibition of protein synthesis under metal stress was found to be a consequence of the inhibition of amino acid uptake. Zn has no effect on amino acid uptake of Silene cells. It is concluded that only Zn tolerance of Silene seems to be related with membrane stability.
Publications

Weiss, M.; Schmidt, J.; Neumann, D.; Wray, V.; Christ, R.; Strack, D.; Phenylpropanoids in mycorrhizas of the Pinaceae Planta 208, 491-502, (1999) DOI: 10.1007/s004250050586

Tissue-specific accumulation of phenylpropanoids was studied in mycorrhizas of the conifers, silver fir (Abies alba Mill.), Norway spruce [Picea abies (L.) Karst.], white pine (Pinus strobus L.), Scots pine (Pinus silvestris L.), and Douglas fir [Pseudotsuga menziesii (Mirbel) Franco], using high-performance liquid chromatography and histochemical methods. The compounds identified were soluble flavanols (catechin and epicatechin), proanthocyanidins (mainly dimeric catechins and/or epicatechins), stilbene glucosides (astringin and isorhapontin), one dihydroflavonol glucoside (taxifolin 3′-O-glucopyranoside), and a hydroxycinnamate derivative (unknown ferulate conjugate). In addition, a cell wall-bound hydroxycinnamate (ferulate) and a hydroxybenzaldehyde (vanillin) were analysed. Colonisation of the root by the fungal symbiont correlated with the distribution pattern of the above phenylpropanoids in mycorrhizas suggesting that these compounds play an essential role in restricting fungal growth. The levels of flavanols and cell wall-bound ferulate within the cortex were high in the apical part and decreased to the proximal side of the mycorrhizas. In both Douglas fir and silver fir, which allowed separation of inner and outer parts of the cortical tissues, a characteristic transversal distribution of these compounds was found: high levels in the inner non-colonised part of the cortex and low levels in the outer part where the Hartig net is formed. Restriction of fungal growth to the outer cortex may also be achieved by characteristic cell wall thickening of the inner cortex which exhibited flavanolic wall infusions in Douglas fir mycorrhizas. Long and short roots of conifers from natural stands showed similar distribution patterns of phenylpropanoids and cell wall thickening compared to the respective mycorrhizas. These results are discussed with respect to co-evolutionary adaptation of both symbiotic partners regarding root structure (anatomy) and root chemistry.
Publications

Neumann, D.; Schwieger, W.; Lichtenberger, O.; Accumulation of Silicon in the Monocotyledons Deschampsia caespitosa, Festuca lemanil and Schoenus nigricans Plant Biol. 1, 290-298, (1999) DOI: 10.1111/j.1438-8677.1999.tb00255.x

Conventional and analytical electron microscopy (EDX, ESI, EELS) were used to investigate the silicon accumulation, the chemical nature of the Si deposits and their formation in three species of monocotyledons. In Deschampsia , in particular parts of the outer epidermal cell wall silicon is accumulated as silicic acid. Electron dense, needle‐shaped crystals in the vacuoles of epidermal cells and in the intercellular spaces were also identified as silicic acid. In xylem parenchyma cells, silicon is accumulated as SiO2, which is formed from Sn silicate. In Festuca , crystal‐like deposits of SiO2 occur on the epidermal surface, in the epidermal and parenchyma cell walls, and in vacuoles of bundle sheath cells. Often the deposits disturb the cell walls and penetrate the envelope of plastids and mitochondria. The crystal‐like SiO2 deposits originate from Sn silicate. In the pericarp of ripe nuts of Schoenus , no stainable cell wall components are detected. The inner part of the pericarp consists of silicic acid, while in the outer regions small clusters of silicic acid are embedded in a matrix of SiO2. The silicic acid deposits show an unusual, layered structure, typical for lepidoic silicic acids, which consist of two‐dimensional crystals lying one above the other.
Publications

Leopold, I.; Günther, D.; Schmidt, J.; Neumann, D.; Phytochelatins and heavy metal tolerance Phytochemistry 50, 1323-1328, (1999) DOI: 10.1016/S0031-9422(98)00347-1

The induction and heavy metal binding properties of phytochelatins in heavy metal tolerant (Silene vulgaris) and sensitive (tomato) cell cultures, in water cultures of these plants and in Silene vulgaris grown on a medieval copper mining dump were investigated. Application of heavy metals to cell suspension cultures and whole plants of Silene vulgaris and tomato induces the formation of heavy metal–phytochelatin-complexes with Cu and Cd and the binding of Zn and Pb to lower molecular weight substances. The binding of heavy metal ions to phytochelatins seems to play only a transient role in the heavy metal detoxification, because the Cd- and Cu-complexes disappear in the roots of water cultures of Silene vulgaris between 7 and 14 days after heavy metal exposition. Free heavy metal ions were not detectable in the extracts of all investigated plants and cell cultures. Silene vulgaris plants grown under natural conditions on a mining dump synthesize low molecular weight heavy metal binding compounds only and show no complexation of heavy metal ions to phytochelatins. The induction of phytochelatins is a general answer of higher plants to heavy metal exposition, but only some of the heavy metal ions are able to form stable complexes with phytochelatins. The investigation of tolerant plants from the copper mining dump shows that phytochelatins are not responsible for the development of the heavy metal tolerant phenotypes.
Publications

Clemens, S.; Kim, E. J.; Neumann, D.; Schroeder, J. I.; Tolerance to toxic metals by a gene family of phytochelatin synthases from plants and yeast EMBO J. 18, 3325-3333, (1999) DOI: 10.1093/emboj/18.12.3325

Phytochelatins play major roles in metal detoxification in plants and fungi. However, genes encoding phytochelatin synthases have not yet been identified. By screening for plant genes mediating metal tolerance we identified a wheat cDNA, TaPCS1 , whose expression in Saccharomyces cerevisiae results in a dramatic increase in cadmium tolerance. TaPCS1 encodes a protein of ∼55 kDa with no similarity to proteins of known function. We identified homologs of this new gene family from Arabidopsis thaliana , Schizosaccharomyces pombe , and interestingly also Caenorhabditis elegans . The Arabidopsis and S.pombe genes were also demonstrated to confer substantial increases in metal tolerance in yeast. PCS‐expressing cells accumulate more Cd2+ than controls. PCS expression mediates Cd2+ tolerance even in yeast mutants that are either deficient in vacuolar acidification or impaired in vacuolar biogenesis. PCS‐induced metal resistance is lost upon exposure to an inhibitor of glutathione biosynthesis, a process necessary for phytochelatin formation. Schizosaccharomyces pombe cells disrupted in the PCS gene exhibit hypersensitivity to Cd2+ and Cu2+ and are unable to synthesize phytochelatins upon Cd2+ exposure as determined by HPLC analysis. Saccharomyces cerevisiae cells expressing PCS produce phytochelatins. Moreover, the recombinant purified S.pombe PCS protein displays phytochelatin synthase activity. These data demonstrate that PCS genes encode phytochelatin synthases and mediate metal detoxification in eukaryotes.
Publications

Bringezu, K.; Lichtenberger, O.; Leopold, I.; Neumann, D.; Heavy Metal Tolerance of Silene vulgaris J. Plant Physiol. 154, 536-546, (1999) DOI: 10.1016/S0176-1617(99)80295-8

Silene vulgaris ssp. humilis, a heavy metal tolerant plant growing on the polluted soil of a medieval copper mining dump, accumulates considerable amounts of heavy metals (HM) in its roots and shoots. The intracellular distribution of HMs in the leaves was investigated by conventional and analytical (EDX, ESI, EELS) electron microscopy. Part of the HMs, Fe, Cu, and Zn occur as crystalline compounds on the surface of the leaves. The epidermal cell walls accumulate Fe, Ni, Cu, AI, Sn, and Zn. Cu within the cell walls is tightly bound to a protein with oxalate oxidase activity, evidencing a high homology to germin. Zn and Sn are accumulated in the cell walls as silicate. Cytoplasm and organelles contain only traces of Cu and Sn, while in the vacuoles no HMs are detected. In the epidermal cell walls, intercellular spaces, and in vacuoles there are high concentrations of Si, forming crystal-like structures. EELS and quantum-chemical calculations reveal these structures as SiO2. The role of Si in the HM-tolerance of Silene is discussed.
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