Substrate specificity and product inhibition of different forms of fructokinases and hexokinases in developing potato tubers

The substrate dependence and product inhibition of three different fructokinases and three different hexokinases from growing potato (Solanum tuberosum L.) tubers was investigated. The tubers contained three specific fructokinases (FK1, FK2, FK3) which had a high affinity for fructose K _m=64, 90 and 100 (M) and effectively no activity with glucose or other hexose sugars. The affinity for ATP (K _m=26, 25 and 240 M) was at least tenfold higher than for other nucleoside triphosphates. All three fructokinases showed product inhibition by high fructose (K _i=5.7, 6.0 and 21 mM) and were also inhibited by ADP competitively to ATP. Sensitivity to ADP was increased in the presence of high fructose, or fructose-6-phosphate. In certain conditions, the K _i (ADP) was about threefold below the K _m (ATP). All three fructokinase were also inhibited by fructose-6-phosphate acting non-competitively to fructose (K _i=1.3 mM for FK2). FK1 and FK2 showed very similar kinetic properties whereas FK3, which is only present at low activities in the tuber but high activities in the leaf, had a generally lower affinity for ATP, and lower sensitivity to inhibition by ADP and fructose. The tuber also contained three hexokinases (HK1, HK2, HK3) which had a high affinity for glucose (K _m=41, 130 and 35 M) and mannose but a poor affinity for fructose (K _m=11, 22 and 9 mM). All three hexokinases had a tenfold higher affinity for ATP (K _m=90, 280 and 560 M) than for other nucleoside triphosphates. HK1 and HK2 were both inhibited by ADP (K _i=40 and 108 M) acting competitively to ATP. HK1, but not HK2, was inhibited by glucose-6-phosphate, which acted non-competitively to glucose (K _i=4.1 mM). HK1 and HK2 differed, in that HK1 had a narrower pH optimum, a higher affinity for its substrate, and showed inhibition by glucose-6-phosphate. The relevance of these properties for the regulation of hexose metabolism in vivo is discussed.Abbreviations FK fructokinase - Fru6P fructose-6-phosphate - Glc6P glucose-6-phosphate - HK hexokinase - NTP nucleoside triphosphate - P_i inorganic phosphate - UDPGlc uridine-5-diphosphoglucose This work was supported by the Deutsche Froschungsgemeinschaft (SFB 137). We are grateful to Professor E. Beck (Lehrstuhl für Pflanzenphysiologie, Universität Bayreuth, FRG) for providing laboratory facilities.     

Über die Bedeutung der oxydo-reduktiven Bedingungen für die vitale Färbung,mit besonderer Berücksichtigung der Kernfärbung in lebendigen Zellen

Ohne ZusammenfassungDie Arbeit wurde angefangen im Peterhofschen Naturwissenschaftlichen Institut.Zum Schlu\ möchte ich Herrn Professor Dr. A. A. Zawarzin, Leiter des Laboratoriums für experimentelle Histologie am Röntgenologischen Institut, und Herrn Dr. D. N. Nassonov, Dozent der Leningrader Universität, für ihre Hilfe und ihr stetes Interesse für meine Arbeit meinen innigsten Dank aussprechen.     

Photosynthetic carbon assimilation in Geosiphon pyriforme (Kützing) F. v. Wettstein,an endosymbiotic association of fungus and cyanobacterium

Geosiphon pyriforme, an endosymbiotic association between a fungus and the cyanobacterium Nostoc, was shown by tracer studies to acquire carbon photosynthetically from CO₂ or bicarbonate. The organism also fixes inorganic carbon in darkness, at lower rates than in the light. The patterns of label distribution are indicative of the operation of the reductive pentose-phosphate pathway in the light and of the phosphoenolpyruvate-carboxylase reaction in the dark. The results are discussed in relation to the evolution of photoautotrophic endosymbiotic associations.This investigation has been supported by the Deutsche Forschungsgemeinschaft. We are indepted to Professor G.B. Feige (Institut für Pflanzenphysiologie, Universität Essen, FRG) for helpful discussion, Karin Faist for her excellent technical assitance and to Doris Schäfer for drawing the figures.     

Beiträge zur Kenntnis der Plasmahaut

Ohne ZusammenfassungDie Arbeit entstand als Dissertation in den Jahren 1922–24 im Botanischen Institut der Universität Leipzig. Herrn Professor Dr.Ruhland sage ich für die Anregung zu der Arbeit und für vielfache Förderung in ihrem weiteren Verlaufe meinen aufrichtigen Dank.     

Vergleichende Permeabilitätsstudien an Pflanzenzellen

Ohne ZusammenfassungHerrn Privatdozent Dr. Czaja, auf dessen Veranlassung die Arbeit im Pflanzenphysiologischen Institut der Universität Berlin in Berlin-Dahlem unternommen wurde, bin ich zu herzlichem Dank für seine fördernden Ratschläge verpflichtet.Ebenso danke ich Herrn Direktor Professor Dr. Noack verbindlich für sein Interesse an der Arbeit und GewÄhrung vieler Hilfsmittel.     

Protoplasmatische Anatomie des Laubmooses Bryum capillare. I

Ohne ZusammenfassungDie Arbeit wurde am Pflanzenphysiologischen Institut der Universität Wien ausgeführt. Dem Vorstand des Institutes Herrn Professor Dr. Karl Höfler erlaube ich mir für die Anregung der Arbeit und die ständige Leitung der Untersuchungen meinen herzlichsten Dank auszusprechen. Für die Einführung in die Fluoreszenzmikroskopie bin ich Herrn Professor Dr. Christian Wimmer (Mödling) zu Dank verpflichtet. Herrn Dr. Lothar Hofmeister danke ich für seine ständige Hilfsbereitschaft herzlichst.     

Soluble acid invertase determines the hexose-to-sucrose ratio in cold-stored potato tubers

Cold storage of potato (Solanum tuberosum L.) tubers is known to cause accumulation of reducing sugars. Hexose accumulation has been shown to be cultivar-dependent and proposed to be the result of sucrose hydrolysis via invertase. To study whether hexose accumulation is indeed related to the amount of invertase activities, two different approaches were used: (i) neutral and acidic invertase activities as well as soluble sugars were measured in cold-stored tubers of 24 potato cultivars differing in the cold-induced accumulation of reducing sugars and (ii) antisense potato plants with reduced soluble acid invertase activities were created and the soluble sugar accumulation in cold-stored tubers was studied. The cold-induced hexose accumulation in tubers from the different potato cultivars varied strongly (up to eightfold). Large differences were also detected with respect to soluble acid (50-fold) and neutral (5-fold) invertase activities among the different cultivars. Although there was almost no correlation between the total amount of invertase activity and the accumulation of reducing sugars there was a striking correlation between the hexose/sucrose ratio and the extractable soluble invertase activitiy. To exclude the possibility that other cultivar-specific features could account for the obtained results, the antisense approach was used to decrease the amount of soluble acid invertase activity in a uniform genetic background. To this end the cDNA of a cold-inducible soluble acid invertase (EMBL nucleicacid database accession no. X70368) was cloned from the cultivar Desirée, and transgenic potato plants were created expressing this cDNA in the antisense orientation under control of the constitutive 35S cauliflower mosaic virus promotor. Analysis of the harvested and cold-stored tubers showed that inhibition of the soluble acid invertase activity leads to a decreased hexose and an increased sucrose content compared with controls. As was already found for the different potato cultivars the hexose/sucrose ratio decreased with decreasing invertase activities but the total amount of soluble sugars did not significantly change. From these data we conclude that invertases do not control the total amount of soluble sugars in coldstored potato tubers but are involved in the regulation of the ratio of hexose to sucrose.The authors are grateful to Heike Deppner and Christiane Prüßner for tuber harvest and technical assistance during the further analysis. We thank Andrea Knospe for taking care of tissue culture, Birgit Schäfer for patient photographic work, Hellmuth Fromme and the greenhouse personnel for attending plant growth and development and Astrid Basner for elucidating the sequence of clone INV-19. The work was supported by the Bundesministerium für Forschung und Technologie (BMFT).     

Zusammenhänge zwischen reduzierenden Eigenschaften und Schutzwirkung von SH-Verbindungen gegen Strahlen

Zusammenfassung Der Abbau der reduzierenden Eigenschaften der Strahlenschutzstoffe Cystein, SH-Glutathion, Thioglykolat und-Monothioglycerin in reinen Lösungen und in Gemischen mit Euglobulin unter dem Einwirken von ultraviolettem Licht und von Röntgenstrahlen wird mittels direkter Jodtitrationen (Stärke als Indikator) bis zum Erliegen der Schutzstoffeigenschaften und darüber hinaus verfolgt.Die Ergebnisse der Versuche mit Cystein, SH-Glutathion und Thioglykolat machen es wahrscheinlich, daß bei diesen drei Verbindungen ein ursächlicher Zusammenhang zwischen reduzierenden Eigenschaften und Schutzstoffeigenschaften besteht.Den Herren Prof. Dr. P. Wels, Dr. F. Markwardt, Dipl.-Chem. H. Dalibor und Apotheker H. D. Schlabitz vom Pharmakologischen Institut sowie Herrn Doz. Dr. G. Henseke vom Institut für Organische Chemie der Universität Greifswald danke ich für fördernde Diskussionen. Fräulein Gerda Rahmel danke ich für technische Assistenz.     

Changing proton concentrations at the surfaces of gravistimulated Phleum roots

The pH patterns at the surfaces of both vertically growing roots of Phleum pratense L. and roots tilted by 45° were recorded using H ⁺-sensitive microelectrodes. During vertical growth the root cap exhibited lower pH values than the meristematic zone. The highest pH values were found at the border between meristematic and elongation zones. In the apical part of the elongation zone the values strongly decreased basipetally. They reached a minimum value of pH 5.4–5.5 (medium pH of about 6.0) at a distance of 700 m from the root tip. This region of strongest acidification usually coincided with that of the highest relative rates of elongation. The region of the first visible curvature following gravistimulation was positioned at 100–200 m more apically. The pH values increased in the basal elongation zone towards the mature zone. The H⁺-flux pattern around a vertically growing Phleum root was characterized by high influxes in the meristematic zone and smaller effluxes in the elongation zone. Tilting the root by 45° induced changes in the pH values of the upper and lower sides of a Phleum root. At a distance of 300–500 m from the root tip, the upper side was strongly acidified while the pH of the lower side slightly increased compared with the values during vertical orientation. pH differences of up to 0.9 pH units between the two sides of a root were detected. These differences decreased basipetally and could not be measured more distant than 700–800 m from the tip. Compared with a vertically growing root, the H⁺-flux pattern of a Phleum root tilted by 45° exhibited effluxes on the entire upper organ flank while the pattern was scarcely altered on the lower side. The curvature-initiating zone in Phleum roots is positioned within that section of the root in which pH changes occur after tilting. The region of highest pH differences, however, is nearer to the apex of the root than the curvature-initiating zone. The pH changes began 8.2 min after a root had been tilted. The bending process started after 17.2 min, i.e. after double the time needed for differential acidification. After reorienting a root, which had just begun to bend, to its previous vertical position the inversion of the pH gradient could be measured within the same mean time of about 8 min. This is again significantly earlier than the beginning of the rebending process. The results indicate that, during the graviresponse, ionic movements occur much earlier than the changes in hormonal activities reported in the literature.Abbreviation CIZ curvature-initiating zone A preliminary report was presented at the 29th Plenary Meeting of the Committee on Space Research (COSPAR) in Washington D.C., USA, 28 Aug – 5 Sept 1992 (Zieschang and Sievers 1993)This work was supported by the Deutsche Forschungsgemeinschaft. We thank Professor H. Felle (Botanisches Institut, Universität Gießen, Gießen, FRG) for practical instructions concerning the method of H⁺-sensitive microelectrodes and Professor W. Simonis (Botanisches Institut, Universität Würzburg, Würzburg, FRG) for allowing to use the microelectrode amplifier.     

Der Einfluss Wasserlöslicher Vitamine auf die Wirksamkeit von Heteroauxin im Wachstumsprozess der Höheren Pflanzen

Ohne ZusammenfassungMit 20 Textabbildungen.Herrn Prof. Dr.Hans Söding gilt mein Dank für das Thema und für sein Interesse an der vorliegenden Arbeit. Die Arbeit wurde durchgeführt im Institut für Allgemeine Botanik der Universität Hamburg. Herrn Prof. Dr.Walter Mevius danke ich für die Erlabnis, diese Arbeit in seinem Institut auszuführen     

Long-term steady-state labelling of wheat plants by use of natural 13CO2/12CO2 mixtures in an open,rapidly turned-over system

A photosynthate labelling method is presented which takes advantage of the natural difference in carbon-isotope composition () which exists between atmospheric CO₂ (-8) and commercially available compressed CO₂. Carbon dioxide with -4.0 and –27.9%., respectively, has been used for labelling. A plant growth cabinet served as the labelling compartment. CO₂-free air was continuously injected at a rate of up to 54m³·h^–1. Dilution of cabinet CO₂ by CO₂-free air was counterbalanced by addition of CO₂ with known constant . Since the labelling-cabinet atmosphere was continuously exchanged at a high rate, photosynthetic carbon-isotope discrimination was fully expressed. In order to study the distribution of carbon acquired by the plant during a defined growth period, the of CO₂ was modified by replacing, for example, atmospheric CO₂ by CO₂ with –27.9%. and the weight and 5 of plant carbon pools was monitored over time. In such an experiment the change of CO₂ was followed by a rapid change of the of sucrose in mature flag-leaf blades of wheat (Triticum aestivum L.). The 5 of sucrose stabilized near –51%., indicating complete exchange by current photosynthate. In contrast 83% of the total carbon in mature flag-leaf blades was not exchanged after 14 d continuous labelling. Differential labelling of pre- and post-anthesis photosynthate indicated that 13% of grain carbon originated from pre-anthesis photosynthesis. Carbon-isotope discrimination and its consideration in experimentation and labelling data evaluation are discussed in detail. Since the air supplied to the labelling cabinet is dry and free of CO₂, carbon-isotope discrimination and carbon turnover and partitioning can be studied over a wide range of CO₂ concentrations (0–2600 cm³ · m^–3) and vapor-pressure deficits.Abbreviation and Symbol PPFD photosynthetic photon flux density - carbon-isotope composition Dr. G. Schleser (Forschungszentrum Jülich, FRG) and Professor S. Hoernes (Mineralogisch-Petrologisches Institut, Universität Bonn) for valuable help and advice during the initial stages of the project and Professor W. Kühbauch (Institut für Pflanzenbau, Universität Bonn) for continuing support. Technical assistance of Ute Labusch, Petra Biermann, Ludwig Schmitz and Thomas Gebbing is gratefully acknowleged.

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Untersuchungen über die Funktion des Zentralnervensystems und insbesondere des Gehirnes bei der Fortbewegung und der Lauterzeugung der Grillen

Ohne ZusammenfassungHerrn Prof. Dr. W. R. Hess in Verehrung und Dankbarkeit gewidmet.Der Deutschen Forschungsgemeinschaft danke ich herzlich für die Förderung dieser Arbeit. Mein Dank gilt ferner den Herren Prof. Dr. O. A. M. Wyss und Dr. R. J. H. Oberholzer, Physiologisches Institut der Universität Zürich, und Herrn Dr. D. Burkhardt, Zoologisches Institut der Universität München, für Beratung in methodischen Fragen.     

Cytologische Untersuchungen an spontanem diploidem Allium carinatum

Ohne ZusammenfassungDie vorliegenden Untersuchungen führte ich im Botanischen Institut der Universität Wien und in der Biologischen Station in Lunz, N.-Österreich, durch. Herrn Prof. Dr. L.Geitler danke ich für das Interesse, das er für meine Untersuchungen gezeigt hat.     

Zur cH-Abhängigkeit der Wasserpermeabilität des Protoplasmas

Ohne ZusammenfassungDie Versuche wurden im Pflanzenphysiologischen Institut der Universität Wien durchgeführt. Es sei gestattet, dem Vorstande des Institutes, meinen verehrten Lehrer, Herrn Prof. Karl Höf1er, für die Anregung der Arbeit und seine ständige Hilfe auch an dieser Stelle den besten Dank auszusprechen. Ebenso danke ich Herrn Prof. Richard Bieb1 für wertvolle Unterstützung bei der Arbeit.     

Assimilation und Atmung von Submersen als Ausdruck ihrer Entquellungsresistenz

Ohne ZusammenfassungDie dieser Abhandlung zugrunde liegenden Versuche wurden in den Jahren 1935 und 1936 im Botanischen Institut der Universität Halle ausgeführt. Die Anregung zu der Arbeit stammt von Herrn Prof. Montfort, dem ich hiermit für das rege Interesse an der Arbeit sowie für die vielseitige Unterstützung meinen besten Dank aussprechen möchte.     

Der Seidenglanz der Kakteenblüten

Ohne ZusammenfassungDie Arbeit wurde im Botanischen Institut der Universität Wien und im II. Physikalischen Institut der Universität Wien in den Jahren 1930 und 1931 ausgeführt. Einige kleine Verbesserungen und Ergänzungen wurden später, vor der Drucklegung, angebracht.An dieser Stelle erlaube ich mir, meinem inzwischen verstorbenen, verehrten Lehrer, Herrn Hofrat Prof. Dr. R v.Wettstein, für die Anregung zu dieser Arbeit meinen Dank auszusprechen.Herrn Prof. Dr. E.Haschek danke ich auf diesem Wege nochmals für sein freundliches Entgegenkommen und die Hilfe, die er mir bei dem physikalisch-optischen Teil meiner Arbeit angedeihen ließ. Durch die Vermittlung Herrn Prof.Hascheks konnte ich über Sommer mit dem vom Physikalischen Institut zur Verfügung gestellten Stufenphotometer bei Frau Direktor V.Prohaska in Klosterneuburg, im Herbst dann im II. Physikalischen Institut arbeiten.Zu Dank verpflichtet bin ich auch Herrn Hofrat Dr. H.Rebel für die Überlassung von seidenglänzenden Schmetterlingen und Herrn Hofrat Dr. K.Keissler für das Entgegenkommen bei der Einsichtnahme in die Sammlung der botanischen Abteilung des Naturhistorischen Museums.Herrn Prof. Dr. H.Cammerloher danke ich herzlichst für die stete Anteilnahme und das intensive Interesse an dem Entstehen der Arbeit.     

Antigenic determinants and specificity of antisera against acidophilic bacteria

Polyclonal antisera, produced against whole cells of Thiobacillus thiooxidans, T. ferrooxidans and Leptospirillum ferrooxidans, gave highly specific reactions when cross-reacted with 23 strains of acidophilic bacteria using an immunofluorescence (IF) staining technique. Strains of identical serotype exhibited maximum cross-reaction whereas strains of different serotype reacted only weakly. Lipopolysaccharides (LPS) examined by SDS-PAGE showed different, serotype-specific migration patterns indicating their rough or smooth character. LPS patterns may therefore be used for serological classification of acidophilic bacteria. Surface antigens of four strains were identified by immunoblot staining; LPS and some proteins were antigenic determinants with LPS the most specific.The authors are with the Universität Hamburg, Institut für Allgemeine Botanik, Abteilung für Mikrobiologie, Ohnhorststrasse 18, D-22609 Hamburg, Germany