Wolfgang Barz

Bio: Wolfgang Barz is an academic researcher from University of Münster. The author has contributed to research in topics: Pterocarpan & Medicarpin. The author has an hindex of 33, co-authored 131 publications receiving 3371 citations.

Photoautotrophic Growth and Photosynthesis in Cell Suspension Cultures of Chenopodium rubrum

Abstract: A method is described for growing cell suspension cultures of Chenopodium rubrum photoautotrophically for prolonged periods of time. By using a two-tier culture vessel the growth medium with the cells was separated from the CO2 reservoir. Definite CO2 concentrations were established by a K2CO3/KHCO3 buffer. Photoautotrophic growth in C. rubrum cell suspension cultures was correlated with the CO2 level. At 0.5% CO2 the cell cultures contained 68 μg chlorophyll/g fresh weight and showed an increase in fresh weight of about 80% in 18 days. At 1% CO2 an increase in fresh weight of 165% in 18 days was observed. The chlorophyll content rose up to 84 μg/g fresh weight. The photoautotrophic growth was also greatly influenced by the 2,4-D content of the medium. Cell growth was enhanced by lowering the auxin concentration. Best growth was attained (210% increase in fresh weight) at 10−8M 2,4-D. The photosynthetic activity of the cells was measured by the light dependent 14CO2 incorporation. At 0.5% CO2 the cell suspensions assimilated about 100 μmol CO2/mg chlorophyll × h. In the presence of 1% CO2 the light driven assimilation was raised up to 185 μmol CO2/mg chlorophyll × h. In both cases, the dark incorporation of CO2 was merely 1.8% of the values obtained in light.

Purification, characterization and differential hormonal regulation of a β -1,3-glucanase and two chitinases from chickpea ( Cicer arietinum L.)

Abstract: Chickpea (Cicer arietinum L.) cell-suspension cultures were used to isolate one beta-1,3-glucanase (EC 3.2.1.29) and two chitinases (EC 3.2.1.14). The beta-1,3-glucanase (M(r) = 36 kDa) and one of the chitinases (M(r) = 32 kDa) belong to class I hydrolases with basic isoelectric points (10.5 and 8.5, respectively) and were located intracellularly. The basic chitinase (BC) was also found in the culture medium. The second chitinase (M(r) = 28 kDa), with an acidic isoelectric point of 5.7, showed homology to N-terminal sequences of class III chitinases and represented the main protein accumulating in the culture medium. Polyclonal antibodies raised against the basic beta-1,3-glucanase (BG) and the acidic chitinase (AC) were shown to be monospecific. The anti-AC antiserum failed to recognize the BC on immune blots, confirming the structural diversity between class I and class III chitinases. Neither chitinase exhibited lysozyme activity. All hydrolases were endo in action on appropriate substrates. The BC inhibited the hyphal growth of several test fungi, whereas the AC failed to show any inhibitory activity. Expression of BG activity appeared to be regulated by auxin in the cell culture and in the intact plant. In contrast, the expression of neither chitinase was apparently influenced by auxin, indicating a differential hormonal regulation of beta-1,3-glucanase and chitinase activities in chickpea. After elicitation of cell cultures or infection of chickpea plants with Ascochyta rabiei, both system were found to have hydrolase patterns which were qualitatively and quantitatively comparable.(ABSTRACT TRUNCATED AT 250 WORDS)

β‐Glucosidases from Cicer arietinum L.

Abstract: β-Glucosidases specific for isoflavone 7-O-glucosides have been isolated from garbanzo plants, Cicer arietinum L. These aryl-β-glucohydrolases occur in the different organs of the plant as multiple molecular forms. The major isoenzymes of the roots, the leaves and the hypocotyl were purified to electrophoretic homogeneity. When subjected to isoelectric focussing in polyacrylamide gels the electrophoretically homogeneous glucohydrolases were found to consist of one or two major and several minor enzymically active molecular species. In roots the β-glucohydrolase isoenzymes constitute a considerable portion of the extractable protein, so that purification to an electrophoretically homogeneous form is easily attainable. All β-glucosidases analyzed possess molecular weights in the range of 125000 (ultracentrifugation) to 135000 (Sephadex G-200) and contain two subunits of molecular weight near 68000. The pH optimum for enzymic activity is 7–7.5 with a second optimum of 4.5–5. The isoelectric points of the various species range between pH 5.9 and 7.1. Staining for glycoprotein was positive. Kinetic analysis demonstrated a pronounced specificity of the enzymes for aromatic substrates with glucose as the sugar moiety. α-Glucosides as well as disaccharides were not hydrolyzed at all. Isoflavone 7-O-glucosides are the most favoured substrates with a Km of 2 × 10−5 M, while the Km with aromatic glucosides (i.e. salicin, 4-nitrophenyl glucoside) are 100 times larger. In addition the β-glucosidases show a pronounced specificity for glucose in the 7-position of the flavonoid nucleus. Using isoflavone aglycones as substrates glucose transferase activity was also demonstrable. The β-glucohydrolase activity is strongly inhibited by Hg2+. This inhibition is partially reversible and preferentially influences the Km values of the enzymes compared to to V. Ag+ glucono-1,5-lactone, ethyleneglycol monomethyl ether and glycerol are only weakly inhibitory, while glucose, p-chloromercuribenzoate and Cu2+ are without effect.

The elicitor-induced oxidative burst in cultured chickpea cells drives the rapid insolubilization of two cell wall structural proteins

Abstract: Elicitation of cultured chickpea cells caused rapid insolubilization of two cell wall structural proteins, p190, a putative hydroxyproline-rich glycoprotein and p80, a putative proline-rich protein. This process appeared to result from an H2O2-mediated oxidative cross-linking mechanism and was initiated within 5 min and complete within 20 min. Further, elicitation of cells induced a rapid, transient generation of H2O2 (oxidative burst), with an onset after 5 min and a maximum H2O2-release after 20 min, as measured by a luminol-dependent chemiluminescence assay. Both chemiluminescence and protein insolubilization were suppressed by exogenous application of catalase or diphenylene iodonium, an inhibitor of plasma-membrane NADPH oxidase, respectively. In contrast, exogenous H2O2 mimicked the effect of the elicitor, suggesting that the putative oxidative crosslinking of the proteins depends directly on H2O2 from the oxidative burst. The peroxidase inhibitor salicylhydroxamic acid blocked both the elicitor- and the exogenous-H2O2-stimulated insolubilization, indicating that a peroxidase activity downstream of H2O2-supply is required. The protein kinase inhibitor staurosporine blocked the elicitation of the oxidative burst and protein insolubilization. In contrast, the protein phosphatase 2A inhibitor cantharidin accelerated, potentiated and extended the elicited oxidative burst. Cantharidin even stimulated the responses in the absence of the elicitor. The competitive effect of both inhibitors confirms that a coordinated activation of (i) protein kinase(s) and (ii) counteracting protein phosphates(s) is a poised signal transduction step for the induction of an NADPH-oxidase-dependent oxidative burst, which drives the putative peroxidase-catalyzed cross-linking of the cell wall proteins.

Stress-Induced Phenylpropanoid Metabolism.

Abstract: Phenylpropanoid compounds encompass a wide range of structural classes and biological functions. Limiting discussion to stress-induced phenylpropanoids eliminates few of the structural classes, because many compounds thst are constitutive in one plant species or tissue can be induced by various stresses in another species or in another tissue of the same plant (Beggs et al., 1987; Christie et al., 1994).

Carbohydrate-modulated gene expression in plants.

Abstract: Plant gene responses to changing carbohydrate status can vary markedly Some genes are induced, some are repressed, and others are minimally affected As in microorganisms, sugar-sensitive plant genes are part of an ancient system of cellular adjustment to critical nutrient availability However, in multicellular plants, sugar-regulated expression also provides a mechanism for control of resource distribution among tissues and organs Carbohydrate depletion upregulates genes for photosynthesis, remobilization, and export, while decreasing mRNAs for storage and utilization Abundant sugar levels exert opposite effects through a combination of gene repression and induction Long-term changes in metabolic activity, resource partitioning, and plant form result Sensitivity of carbohydrate-responsive gene expression to environmental and developmental signals further enhances its potential to aid acclimation The review addresses the above from molecular to whole-plant levels and considers emerging models for sensing and transducing carbohydrate signals to responsive genes

Visualization of protein interactions in living plant cells using bimolecular fluorescence complementation.

Abstract: Dynamic networks of protein-protein interactions regulate numerous cellular processes and determine the ability to respond appropriately to environmental stimuli. However, the investigation of protein complex formation in living plant cells by methods such as fluorescence resonance energy transfer has remained experimentally difficult, time consuming and requires sophisticated technical equipment. Here, we report the implementation of a bimolecular fluorescence complementation (BiFC) technique for visualization of protein-protein interactions in plant cells. This approach relies on the formation of a fluorescent complex by two non-fluorescent fragments of the yellow fluorescent protein brought together by association of interacting proteins fused to these fragments (Hu et al., 2002). To enable BiFC analyses in plant cells, we generated different complementary sets of expression vectors, which enable protein interaction studies in transiently or stably transformed cells. These vectors were used to investigate and visualize homodimerization of the basic leucine zipper (bZIP) transcription factor bZIP63 and the zinc finger protein lesion simulating disease 1 (LSD1) from Arabidopsis as well as the dimer formation of the tobacco 14-3-3 protein T14-3c. The interaction analyses of these model proteins established the feasibility of BiFC analyses for efficient visualization of structurally distinct proteins in different cellular compartments. Our investigations revealed a remarkable signal fluorescence intensity of interacting protein complexes as well as a high reproducibility and technical simplicity of the method in different plant systems. Consequently, the BiFC approach should significantly facilitate the visualization of the subcellular sites of protein interactions under conditions that closely reflect the normal physiological environment.

Secondary metabolites in plant defence mechanisms

Abstract: SUMMARY Many secondary metabolites found in plants have a role in defence against herbivores, pests and pathogens. In this review, a few examples are described and discussed, and some of the problems in determining the precise role(s) of such metabolites highlighted. The role of secondary metabolites in defence may involve deterrence/anti-feedant activity, toxicity or acting as precursors to physical defence systems. Many specialist herbivores and pathogens do not merely circumvent the deterrent or toxic effects of secondary metabolites but actually utilize these compounds as either host recognition cues or nutrients (or both). This is true of both cyanogenic glucosides and glucosinolates, which art discussed in detail as examples of defensive compounds. Their biochemistry is compared and contrasted. An enormous variety of secondary metabolites are derived from shikimic acid or aromatic amino acids, many of which have important roles in defence mechanisms. Several classes of secondary products are ‘induced’ by infection, wounding or herbivory, and examples of these are given. Genetic variation in the speed and extent of such induction may account, at least in part, for the difference between resistant and susceptible varieties. Both salicylates and jasmonates have been implicated as signals in such responses and in many other physiological processes, though their prescise roles and interactions in signalling and development are not fully understood.

The functions and regulation of glutathione s-transferases in plants.

Abstract: ▪ Abstract Glutathione S-transferases (GSTs) play roles in both normal cellular metabolism as well as in the detoxification of a wide variety of xenobiotic compounds, and they have been intensively studied with regard to herbicide detoxification in plants. A newly discovered plant GST subclass has been implicated in numerous stress responses, including those arising from pathogen attack, oxidative stress, and heavy-metal toxicity. In addition, plant GSTs play a role in the cellular response to auxins and during the normal metabolism of plant secondary products like anthocyanins and cinnamic acid. This review presents the current knowledge about the functions of GSTs in regard to both herbicides and endogenous substrates. The catalytic mechanism of GST activity as well as the fate of glutathione S-conjugates are reviewed. Finally, a summary of what is known about the gene structure and regulation of plant GSTs is presented.