Dependence of the detectability of stored growth on the elongation rate in IAA- and acid-induced elongation of wheat coleoptile sections

The detectability of stored growth at various elongation rates(IAA- and acid- induced) was investigated in 5-mm wheat coleoptilesegments. After 20 min turgor reduction by 0.15 M or 0.20 Mmannitol, the detectability of stored growth depended on theelongation rate before turgor reduction. A hypothesis was proposedthat the amount of stored growth is limited. Depending on theelongation rate, this then appears as complete or partial compensationof the growth lost during turgor reduction. The limit for fullcompensation was about 300 µm/hr?segment. At elongationrates of > 600 µm/hr?segment, no stored growth couldbe detected. The elongation of the wheat coleoptile sectionsat low and high elongation rates is assumed to be limited bydifferent rate-determining steps. (Received November 22, 1977; )     

Isolation of chick tenascin variants and fragments. A C-terminal heparin-binding fragment produced by cleavage of the extra domain from the largest subunit splicing variant.

The extracellular-matrix glycoprotein, tenascin, consists of disulfide-linked subunits of 190, 200 and 230 kDa (the three splicing variants reported in chicken) and usually exists as a six-armed structure under the electron microscope. We used monoclonal antibodies to isolate and characterize different splicing variants and proteolytic fragments obtained from the native protein. Purified monomeric tenascin has a native molecular mass of 216 kDa and is structured as single arms. Tenascin fragments obtained by pepsin digestion bind to monoclonal antibody (mAb) TnM1 which is directed against epidermal-growth-factor-like repeats in the N-terminal half of all subunits. These fragments represent the thin proximal part of the tenascin arms and they are still partially linked to dimers and trimers via disulfide bridges. Using mAb Tn68, that reacts with a fibronectin-type-III repeat towards the C-terminus, a tenascin fragment, generated by treatment with pronase, can be isolated. Ultrastructurally, this fragment looks like the thicker distal part of the tenascin arms. Only the 230-kDa variant of tenascin gives rise to this distal fragment after cleavage within the alternatively spliced fibronectin-type-III repeats. Native tenascin and all fragments containing the distal part of its arms bind to heparin-agarose, whereas the proximal fragments do not. Oligomeric and monomeric tenascin inhibit fibronectin-mediated fibroblast spreading with comparable efficiency when added to the culture medium, while the proximal fragment has no effect. The distal fragment as well as reduced and alkylated tenascin are active in this assay, but only at higher molar concentrations when compared to the native protein.     

Improved method for electroporation of Staphylococcus aureus

We have developed a significantly improved method for the electroporation of plasmid DNA into Staphylococcus aureus. The highest transformation efficiency achieved with this procedure was 4.0 x 10(8) transformants per microgram of plasmid pSK265 DNA. This represents a 530-fold improvement over the previously reported optimum efficiency of 7.5 x 10(5) transformants per microgram of plasmid DNA after electroporation of S. aureus cells [9]. Identical results were obtained when electrocompetent cells, which had been stored frozen at -80 degrees C, were used. The improved efficiency is due primarily to the use of a modified medium (designated as B2 medium) and secondarily to the use of 0.1-cm cuvettes. Several other plasmids (pI258, pMH109, and pSK270) were also electrotransformed into competent cells using our procedure, and for each plasmid, the transformation efficiency was significantly reduced compared to that observed when pSK265 DNA was used. With respect to plasmid pI258, the transformation efficiency was 3500-fold higher than that reported previously for transformation of this plasmid into S. aureus RN4220 [9]. The optimized electroporation procedure was less successful in transforming other staphylococci. Electrocompetent cells of S. aureus ATCC 29213 and S. epidermidis ATCC 12228 produced 5.5 x 10(5) and 5 x 10(3) transformants per microgram of pSK265 DNA, respectively.     

Ferredoxin:NADP oxidoreductase of Cyanophora paradoxa: purification, partial characterization, and N-terminal amino acid sequence.

The ferredoxin:NADP+ oxidoreductase of the protist Cyanophora paradoxa, as a descendant of a former symbiotic consortium, an important model organism in view of the Endosymbiosis Theory, is the first enzyme purified from a formerly original endocytobiont (cyanelle) that is found to be encoded in the nucleus of the host. This cyanoplast enzyme was isolated by FPLC (19% yield) and characterized with respect to the uv-vis spectrum, pH optimum (pH 9), molecular mass of 34 kDa, and an N-terminal amino acid sequence (24 residues). The enzyme shows, as known from other organisms, molecular heterogeneity. The N-terminus of a further ferredoxin:NADP+ oxidoreductase polypeptide represents a shorter sequence missing the first four amino acids of the mature enzyme.     

Der Einfluss der Temperatur Auf die Lage des CO2-Kompensationspunktes

Zusammenfassung Der Begriff CO₂-Kompensationspunkt wird im Vergleich zum Begriff Licht-Kompensationspunkt an Hand von Meßergebnissen erläutert. Die Lage des CO₂-Kompensationspunktes ist von der Temperatur abhängig derart, daß sich bei höherer Temperatur das Gleichgewicht zwischen CO₂-Aufnahme und-abgabe im belichteten Gewebe bei einem höheren CO₂-Partialdruck der Umgebung einstellt. Die Temperaturkoeffizienten für die Dunkelatmung, die apparente und die gesamte Photosynthese werden in Form vonQ ₁₀-Werten wiedergegeben. Da diese Werte bei der photosynthetischen CO₂ Verarbeitung wesentlich kleiner sind wie die bei photochemischen und chemischen Reaktionen beobachteten Temperaturkoeffizienten, wurde vermutet, daß der mit zunehmender Temperatur beobachtete steile Abfall der apparenten Photosynthese sowie die Temperaturabhängigkeit der Gesamtphotosynthese in Form einer Optimumkurve auf eine temperaturabhängige Veränderung der Reaktionsbedingungen zurückzuführen ist. Solche temperaturbedingten Veränderungen sind in bezug auf das CO₂-Diffusionsgefälle zwischen der die Pflanze umgebenden Luft und den Reaktionsorten für die Bindung und photosynthetische Verarbeitung des Kohlendioxyds im Gewebe gegeben, so daß anzunehmen ist, daß die beobachteten Temperaturwirkungen in erster Linie auf temperaturbedingte Veränderungen physikalisch-physiologischer Größen bei dem vor der eigentlichen photochemischen und chemischen CO₂-Verarbeitung stattfindenden Diffusionsvorgang zurückzuführen sind.Mit 8 Textabbildungen.Herrn Prof. Dr.O. Renner zum 70. Geburtstag gewidmet.