Weitere Untersuchungen zur phytochrominduzierten Akkumulation von Ascorbinsäure beim Senfkeimling (Sinapis alba L.)

Summary The accumulation of ascorbic acid (AS) in the young mustard seedling is greatly increased by the action of the active form of phytochrome (P₇₃₀) (see Schopfer, 1966). There is no photosynthesis in continuous far-red light, which was used throughout the experiments. The phytochrome-mediated increase in AS approximately parallels the synthesis of anthocyanin in the seedling, although the onset of AS-accumulation precedes the anthocyanin synthesis by 2–3 hours (Fig. 4 and 5).—The action of P₇₃₀ increases the amount of AS in every part of the seedling (cotyledons, hypocotyl, radicula) (Fig. 1–3). This increase in AS parallels the formation of P₇₃₀ rather than the different growth responses of these organs (enlargement of the cotyledons, inhibition of hypocotyl and radicula lengthening). The lag in AS-accumulation after onset of far-red irradiation is the same in all 3 parts of the seedling (about 1 hour under the experimental conditions; Fig. 4).—Actinomycin D (10 g/ml), which strongly inhibits anthocyanin synthesis (Fig. 9 and 10), has no corresponding effect on P₇₃₀-dependent increase in AS-accumulation (Fig. 7 and 8). This result support the hypothesis that already active genes are only slightly influenced by actinomycin D (see Lange and Mohr, 1965). It also shows that, in contrast to its role in anthocyanin synthesis, P₇₃₀ probably does not act by initiating potentially active genes in the case of AS-accumulation. — A dark synthesis of anthocyanin in the cotyledons can be obtained by application of AS to the seedlings (Fig. 11). Glucose and sucrose are ineffective in this respect (Table 2). The effect of AS-feeding on anthocyanin synthesis can be inhibited by actinomycin D in very much the same way as light induced anthocyanin synthesis is inhibited (Table 3). — Also the RNA content of the cotyledons is increased by feeding AS in the dark (Table 4).These results are in line with the earlier suggested hypothesis (see Schopfer, 1966) that increase in AS-accumulation is a very early event in the mediation of some positive photoresponses, e.g. anthocyanin synthesis. According to the hypothesis of Mohr (1966 a, b) it has been concluded that AS functions as a part of the signal chain of phytochrome-mediated photomorphogenesis. This signal chain is thought to start differential gene activation to bring about positive photoresponses.     

Vom Einfluß des Phytochroms auf die Xylemdifferenzierung im Hypokotyl des Senfkeimlings (Sinapis alba L.)

Zusammenfassung P₇₃₀, das aktive Phytochrom, bewirkt eine vermehrte Bildung von Gefäßen (Tracheen und Tracheiden) im Hypokotyl des Senfkeimlings. Das Differenzierungsmuster der Leitbündel und der Verlauf der Leitbahnen sind im belichteten und im etiolierten Keimling gleich. Es wird geschlossen, daß auch bezüglich der Ausbildung der Leitbahnen das P₇₃₀ lediglich im Rahmen einer sekundären Differenzierung als Auslöser wirkt. Die primäre Differenzierung (vgl. Wagner und Mohr, 1966 b) wird durch P₇₃₀ offenbar nicht beeinflußt.

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Phytochrome-mediated control of xylem differentiation in the hypocotyl of the mustard seedling (Sinapis alba L.)

Summary P₇₃₀, the active phytochrome, causes an increased formation of xylem elements (tracheids and vessel elements) in the hypocotyl of the mustard seedling (Figs. 3,4). On the other hand, the pattern of differentiation of the bundles and the course of the bundles within the hypocotyl (Figs. 1,2) are the same in etiolated as well as in illuminated seedlings.—It has been concluded that in connection with bundle differentiation P₇₃₀ acts only as a trigger at the level of secondary differentiation. The pattern of differentiation is laid down in the course of primary differentiation which apparently is not influenced by P₇₃₀. The same problem has been dealt with more in detail in a foregoing paper (Wagner and Mohr, 1966b).

     

Die Wirkung von Actinomycin D auf die phytochromabhängigen Veränderungen des RNS-Gehaltes im Senfkeimling (Sinapis alba L.)

Summary Actinomycin D (10 g/ml) cancels completely the phytochrome-mediated RNA net synthesis in the cotyledons of the mustard seedling whereas RNA net synthesis in the cotyledons of the dark-grown seedling is only partially inhibited (Fig. 1). — In the hypocotyl Actinomycin D of the same concentration lowers the RNA contents in the light (i.e. far-red)-grown seedling as well as in the dark-grown seedling down to the same level (Fig. 2). In the presence of Actinomycin D phytochrome has no significant influence on the RNA contents neither in the cotyledons nor in the hypocotyl (Fig. 1,2).The data support the view that P₇₃₀, the active phytochrome, acts through differential gene activation in the cotyledons and predominantly through differential gene repression in the hypocotyl (cf. Mohr, 1966; Schopfer, 1967a, b). —The data further support the conception that active genes (as defined by Mohr, 1966 and Schopfer, 1967a, b) are much less sensitive towards Actinomycin D than potentially active and repressible genes (cf. Schopfer, 1967a; Mohr and Bienger, 1967).     

Die Hemmung der Phytochrominduzierten Photomorphogenese („Positive” Photomorphosen) des Senfkeimlings (Sinapis alba L.) Durch Actinomycin D und Puromycin

Zusammenfassung Die positiven Photomorphosen Öffnung des Hypokotylhakens und Entfaltung der Kotyledonen können ganz ähnlich wie die phytochrominduzierte Anthocyansynthese und andere positive Photomorphosen durch Actinomycin D und Puromycin gehemmt werden. Man kann daraus schließen, daß diese beiden photomorphogenetischen Reaktionen des Senfkeimlings ebenfalls durch eine von P₇₃₀ über eine Signalkette ausgelöste Aktivierung von potentiell aktiven Genen veranlaßt werden.

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The inhibition of phytochrome-mediated photomorphogenesis (positive photoresponses) by actinomycin D and puromycin in the mustard seedling (Sinapis alba L.)

Summary The many photochrome-mediated photoresponses of a seedling (Sinapis alba L., white seeded mustard) can be divided into 3 categories: positive, negative, and complex photoresponses. Positive photoresponses are those which are characterized by an initiation or a promotion of biosynthetic or growth processes (Mohr, 1966b). Phytochrome-mediated anthocyanin synthesis is the prototype of a positive photoresponse. It has been shown in previous papers (e.g. Lange and Mohr, 1965; Mohr et al., 1965) that positive photoresponses can be specifically inhibited by actinomycin D and puromycin. It has been concluded that in the case of positive photoresponses P₇₃₀ (the active phytochrome) exerts its function through differential gene activation.—In the present paper it has been demonstrated that phytochrome-mediated positive photoresponses of the mustard seedling like opening of the hypocotylar hook and unfolding of the cotyledons can be inhibited by relatively low doses of actinomycin D and puromycin in very much the same way as anthocyanin synthesis or cotyledon enlargement is inhibited. It has been concluded that in these cases too the action of P₇₃₀ must be attributed to an activation of potentially active genes in the manner postulated on the basis of the data on anthocyanin synthesis.

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Die Arbeit wurde durch Sachbeihilfen der Deutschen Forschungsgemeinschaft und der Stiftung Volkswagenwerk (an Prof. Mohr) ermöglicht.     

Zur Ei- und Embryonalentwicklung des HydroidpolypenEudendrium armatum

Zusammenfassung 1. Die Oocyten-, Blastostyl- und Embryonalentwicklung vonEudendrium armatum Tichomirov wurde licht- und elektronenmikroskopisch untersucht.2. Oocyten entstehen einzeln oder in dichter Lagerung aus undifferenzierten Zellen des Ektoderms in jüngeren sowie älteren Hydrocaulusabschnitten. Bereits vor der Blastostylknospung sind im Hydrocaulus zahlreiche Oocyten vorhanden. Gesetzmäßige Lagebeziehungen zwischen den Orten der Oocytenentstehung (Keimzonen) und dem Verzweigungssystem lassen sich nicht feststellen. Das Wandern der Oocyten im Hydrocaulus kann am lebenden Stöckchen verfolgt werden.3. Blastostyle sind von Nährpolypen im Knospenzustand durch in ihren Gastralraum eingewanderte Oocyten und später durch ihre Spadixbildung unterscheidbar. Die möglichen Wechselbeziehungen zwischen Oogenese und Oocytenwanderung einerseits und Blastostylen andererseits werden diskutiert.4. Während der Vitellogenese wird vom Spadixentoderm granulöses Material — möglicherweise Glykogen — an die Oocyte abgegeben. Das Spadixentoderm hat durch Zellausläufer direkten Kontakt mit der Oocyte.5. Nach der Befruchtung bildet die Oocyte eine Eihülle. Das Material dieser Eihülle entspricht wahrscheinlich der Peridermsubstanz.6. Die Eihülle wird gleichzeitig mit dem Periderm unterhalb des Blastostyls verlötet. Dies geschieht in Wechselbeziehung zu einer Abhebung und Retraktion des Spadix vom Ei. Danach bleibt die Stützlamella aus dem Spadixbereich als gefaltetes Paket in der Gastralwand des Blastostyls liegen. Der fibrilläre Randsaum ist weitgehend ungestört. Es wird diskutiert, ob die Myoepithelzellen ihre Bindung an die Stützlamelle lösen und gegebenenfalls wieder knüpfen können.7. Die Furchung verläuft — zumindest vom 8-Kern-Stadium ab — total. Der Beginn der Durchfurchung wurde stets in zeitlicher Verzögerung zu den ersten Kernteilungen beobachtet. Zellgrenzen wurden frühestens im 4-Kern-, spätestens im 8-Kern-Stadium gefunden. Die widersprüchlichen Angaben über totale, syncytiale und superfizielle Furchung in der GattungEudendrium werden an Hand der Befunde diskutiert.8. Am Ende der Vitellogenese und zu Beginn der Embryonalentwicklung werden homogene Dotterbereiche in — je nach Fixierung unterschiedlicher — Verbreitung gefunden. In solchen Verflüssigungsbereichen liegt der Komplexdotter nicht in von Membranen umgrenzten Tröpfchen (Vesikeln) vor. Diese Bildung wird als Fixierungsartefakt gedeutet.9. Die histologische Differenzierung beginnt bereits in der späten Furchung parallel zur Anlage der Körperschichten. Der beschriebene Entstehungsmodus der Zweischichtigkeit kann als Moruladelamination bezeichnet werden.

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On egg and embryonic development of the hydroid polypEudendrium armatum. A light and electron microscopic study

Oocytes ofEudendrium armatum originate in the branching stalks from undifferentiated ectoderm cells, in younger as well as in older parts, individually or in groups — prior to the development of blastostyles. Oocyte migration is caused by an autonomous activity. Possible interrelationships between oogenesis and the migration of oocytes on the one hand, and the development of gonozoids on the other are discussed. Cleavage is complete, at least from the eighth nucleus stage onwards. Controversial opinions about cleavage in various species ofEudendrium are discussed, with special reference to the problem of the syncytial cleavage and areas of liquefied yolk. InE. armatum, the latter is regarded as an artefact of fixation. Egg shell formation, function and retraction of the spadix and embryogenesis are described.

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Abkürzungen in den Abbildungen B Bakterium - Bl Blastostyl - Cb Cnidoblast - Ci Cilium - Ch Chromatin - dEhG dunkle Eihüllen-Grana - Dm Doppelmembran - Do Komplexdotter - Drg Drüsenring - Drz Grana-haltige Drüsenzellen - Eh Eihüllenschicht I - Eh Eihüllenschicht II - Ek Ektoderm - Em Extrusionsmaterial - Em vermutliches Em, von Doppelmembran umgeben - En Entoderm - F Filamente - Fa Fixierung nachFahrenbach - f.Ch fibrilläres Chromatin - Fp Freßpolyp - F.Stl Fibrillen der Stützlamelle - Fs.Stl Fibrillensaum der Stützlamelle - -C glatte Membranen - G Gastralraum - Gl vermutliches Glycogen - Go Golgi-Apparat - Gr strukturiertes Granum - hEhG helle Eihüllen-Grana - I-Z I-Zelle - k.Ch kondensiertes Material innerhalb des fibrillären Chromatin - Mf Myofibrillen - Mi OsO₄ nachMillonig - Mit Mitochondrium - N Nucleus - Nl Nucleolus - Nm Kernmembran - Np Kernporen - O Oocyte - Pa OsO₄ nachPalade - Pd' äußeres Periderm - Pdb Peridermbildungszone - PdG Peridermbildungs-Grana - hPdG helle Peridermbildungs-Grana - dPdG dunkle Peridermbildungs-Grana - p.f pars fibrosa des Nucleolus - p.g pars granulosa des Nucleolus - Psp pseudopodienartiger Ausläufer einer Oocyte - R Ribosomen - Sp Spadix - Sp.R Spadix in Retraktion - Stl Stützlamelle - StM Styrol Methacrylat - Sz Schleimzellen - V Vesikel und V-reihen - /V Vestopal - Verfl Dotter-Verflüssigungszone - Wp Wehrpolyp - X peripherer Bereich ohne Zellorganelle - Zm Zellmembran     

Die hemmung der Phytochrom-induzierten Anthocyansynthese durch puromycin und 2-thiouracil

Zusammenfassung Die durch Phytochrom 730 bewirkte Anthocyansynthese des Senfkeimlings kann durch Puromycin und 2-Thiouracil gehemmt werden. Im Gegensatz zu Actinomycin D (Lange u Mohr, 1965) hemmen diese Substanzen die Anthocyansynthese auch dann fast völlig, wenn sie erst längere Zeit, z. B. 12 Std. nach Lichtbeginn appliziert werden. Der Effekt von 2-Thiouracil kann aufgehoben werden, wenn man die Keimlinge in ein Medium mit Uracil oder Wasser bringt. Auch die gleichzeitige Applikation von Uracil und 2-Thiouracil führt zu einer zumindest partiellen Aufhebung des Hemmeffektes von 2-Thiouracil. Aus den Kinetiken kann man folgern, daß die Lebensdauer des kurzlebigsten Enzyms, das an der Anthocyansynthese beteiligt ist, in der Größenordnung von 6 Std liegt. Der Schluß erscheint aufgrund der Abb. 1. u. 2 berechtigt, daß die Synthese dieses Enzyms über P₇₃₀ reguliert wird. — Die Wirkung von 2-Thiouracil kann in unserem Fall offensichtlich nicht nur damit gedeutet werden daß 2-Thiouracil in RNS engebaut wird und dadurch falsche RNS entsteht. Man muß vielmehr annehmen, daß das 2-Thiouracil auch eine direkte, kompetitive Enzymhemmung verursacht.

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The inhibitory effect of puromycin and 2-thiouracil on the phytochrome-mediated synthesis of anthocyanin

Summary The phytochrome-mediated anthocyanin synthesis of the mustard seelding, Sinapis alba L., can be stopped by the application of relatively low concentrations of puromycin and 2-thiouracil. While actinomycin D, which has been investigated in a previous paper (Lange and Mohr, 1965), will block phytochrome-induced anthocyanin synthesis only if it is applied before or at the onset of light, puromycin and 2-thiouracil will stop anthocyanin synthesis even if they are applied at a later stage, e.g., 12 hours after the onset of light. — The effect of 2-thiouracil can be reversed if the seedlings are transferred to a medium containing uracil or to blank water. The simultaneous application of 2-thiouracil and uracil also leads to at least a partial reversal of the 2-thiouracil effect. — The kinetics of the puromycin inhibition (Fig. 2) indicate that the enzyme with the shortest life-time among those enzymes which are involved in anthocyanin production has a life-time in the order of 6 hours. One may reasonably conclude on the basis of Fig. 1 and 2 that synthesis of this particular enzyme is controlled by phytochrome 730. — The effect of 2-thiouracil (Fig. 3) cannot — in our case at least — be understood by only assuming that 2-thiouracil will be incorporated into RNA and thus will lead to the formation of wrong RNA. Inhibition by 2-thiouracil is much faster than inhibition by puromycin (Fig. 2,3). We have rather to conclude that 2-thiouracil may exert its effect mainly through a direct competitive enzyme inhibition.

     

Der Einfluss von Phytochrom auf die Stationären Konzentrationen von Ascorbinsäure und Dehydroascorbinsäure beim Senfkeimling (Sinapis alba L.)

Zusammenfassung Der Senfkeimling (Sinapis alba L.) synthetisiert auch im Dunkeln beträchtliche Mengen an Ascorbinsäure. Durch Licht kann der Ascorbinsäure-Gehalt der Keimlinge stark erhöht werden. Dieser Lichteinfluß ist auf die Funktion von Phytochrom zurückzuführen. Die Photosynthese hat keinen wesentlichen Anteil an der Lichtwirkung. Die Konzentration an Dehydroascorbinsäure ist im Verhältnis zur Ascorbinsäurekonzentration stets sehr gering (5–8% der Konzentration an Totalascorbat) und wird vom Phytochrom nicht beeinflußt.Wenn die Funktion von Phytochrom bei den positiven Photomorphosen unter dem Aspekt der differentiellen Genaktivierung (vgl. Mohr, 1966) betrachtet wird, kann die folgende Arbeitshypothese aufgestellt werden: Die durch Phytochrom 730 induzierte Ascorbinsäureakkumulation führt im Bereich von potentiell aktiven Genen zu einer Spaltung gewisser DNS-Histon-Komplexe. Dadurch wird die Synthese von m-RNS an diesen Genen ermöglicht, was schließlich zur Ausbildung der positiven Photomorphosen führt. Die Argumente, welche zur Zeit für eine Funktion der Ascorbinsäure bei der Regulation der Genaktivität sprechen, werden kurz diskutiert.

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The control by phytochrome of the contents of ascorbic acid and dehydroascorbic acid in the mustard seedling (Sinapis alba L.)

Summary A dark grown seedling of white seeded mustard (Sinapis alba L.) contains an appreciable amount of ascorbic acid. The content of ascorbic acid, however, will strongly increase under the influence of light. This effect is due to phytochrome. Photosynthesis is not involved under our experimental conditions.The content of dehydroascorbic acid is always very low compared to ascorbic acid (5–8% of total ascorbate). Phytochrome does not influence this relation.The lag-phase of the phytochrome induced increase in ascorbic acid accumulation is remarkably short, about 1 hour after the onset of light compared to about 4 hours for phytochrome induced anthocyanin synthesis under our conditions.This is the shortest lag-phase we have observed hitherto in the case of positive photoresponses (Mohr, 1966).If we assume that the function of phytochrome in the case of positive photoresponses involves a differential gene activation of potentially active genes (Mohr, 1966) the following working hypothesis can be advanced: phytochrome induced accumulation of ascorbic acid will lead to a separation of DNA-histone complexes in the range of potentially active genes. This makes possible the DNA-dependent synthesis of m-RNAs at those sites which are lastly responsible for the initiation of positive photoresponses. — Arguments are briefly considered which support the view that ascorbic acid exerts a function in connection with the regulation of gene activity.

     

Purification and characterization of flavone-specific β-glucuronidase from callus cultures of Scutellaria baicalensis Georgi

-Glucuronidase from callus cultures of Scutellaria baicalensis Georgi was purified to apparent homogeneity by fractionated ammonium-sulfate precipitation and chromatography on diethylaminoethyl-cellulose, hydroxylapatite and baicalin-conjugated Sepharose 6B. A 650-fold purification was obtained by this purification system. When subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified protein migrated as a single band with a molecular mass of 55 kDa. We determined that the native enzyme has a molecular mass of 230 kDa using gel-filtration chromatography. These results suggested that the enzyme exists as a homotetramer composed of four identical 55-kDa subunits. The enzyme showed a broad pH optimum between 7.0 and 8.0. The K _m values were 9 M, 10 M, 30 M and 40 M for luteolin 3 -O--d-glucuronide, baicalin, wogonin 7-O--d-glucoronide and oroxlin 7-O--d-glucuronide, respectively. The enzyme was most active with flavone 7-O--d-glucuronides.Abbreviations BA N⁶-benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - pI isoelectric point - R _t retention time     

Abbau von 14C-, 3H- und 36Cl-markiertem γ-Hexachlorcyclohexan durch anaerobe Bodenmikroorganismen

Zusammenfassung Durch eine anaerobe Mischflora aus Ackerboden wurde -Hexachlorcyclohexan (-HCH) in 4–5 Tagen zu 90% abgebaut. Dabei erfolgte eine schnelle Abspaltung des Chlors in Form von Chloridionen und danach eine Freisetzung des C- und H-Anteiles in Form flüchtiger Verbindungen, in denen kein Chlor und auch kein CO₂ nachzuweisen war.Die Verwendung von ¹⁴C/³H- und ³⁶Cl/³H-doppelmarkiertem -HCH zeigte, daß die Cl- und H-Abspaltung nicht im Verhältnis von 1:1 erfolgte, sondern mehr Cl als H abgespalten wurde. Die flüchtigen Verbindungen enthielten andererseits höhere ¹⁴C- als ³H-Anteile. Gaschromatographische Untersuchungen zeigten ebenfalls eine rasche Verminderung des -HCH und die Bildung verschiedener Metabolite. Es wurde jedoch kein -Pentachlorcyclohexen nachgewiesen. Bei steigenden O₂-Gehalten in der Gasphase verminderte sich der -HCH-Abbau. Jedoch fanden auch noch bei 5% O₂ Chlorabspaltung und die Freisetzung flüchtiger Metabolite statt.-HCH wurde ebenfalls, jedoch langsamer, durch die anaerobe Mischflora abgebaut. Auch hier wurde Chlorid abgespalten, und es traten ebenfalls flüchtige Verbindungen auf, die kein Chlor enthielten.

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Degradation of ¹⁴C-, ³H- and ³⁶Cl-labelled -hexachlorocyclohexane by anaerobic soil microorganisms

Up to 90% of the -Hexachlorocyclohexane (-HCH) applied to an anaerobic mixed bacterial flora enriched from an arable soil were degraded within 4–5 days. Degradation resulted in a rapid release of chloride and in formation of chlorine-free volatile metabolites. CO₂ formation from the molecule was not detected.Investigations with ¹⁴C/³H- and ³⁶Cl/³H double-labelled -HCH indicated that the release of Cl and H did not occur in the ratio of 1:1. More Cl than H was split off. The volatile compounds contained more ¹⁴C than ³H. Gas chromatographic studies also showed the rapid decrease of -HCH and the formation of several metabolites. -Pentachlorocyclohexene was not detected. Increasing O₂-contents in the gas phase of cultures resulted in decreases of the compound's degradation. Release of chloride and of volatile metabolites were observed with O₂ contents in the gas phase up to 5%.-HCH was also, but more slowly as with -HCH, degraded by the anaerobic mixed flora. Chloride was released and volatile, chlorine-free metabolites were found.

     

Isolation and characterization of β-glucosidases from Aspergillus nidulans mutant USDB 1183

Two extracellular -glucosidases (cellobiase, EC 3.2.1.21), I and II, from Aspergillus nidulans USDB 1183 were purified to homogeneity with molecular weights of 240,000 and 78,000, respectively. Both hydrolysed laminaribiose, -gentiobiose, cellobiose, p-nitrophenyl--L-glucoside, phenyl--L-glucoside, o-nitrophenyl--L-glucoside, salicin and methyl--L-glucoside but not -linked disaccharides. Both were competitively inhibited by glucose and non-competitively (mixed) inhibited by glucono-1,5-lactone. -Glucosidase I was more susceptible to inhibition by Ag⁺ and less inhibited by Fe²⁺ and Fe³⁺ than -glucosidase II.     

Bewegungsstudien an Anatinen

Zusammenfassung Vorliegende Arbeit ist eine Weiterführung derLorenz'schen Bewegungsstudien an Anatinen aus dem Jahre 1941, fortgesetzt an Mischlingen zwischen den dort beschriebenen Arten. Die sich dabei ergebenden Befunde machten eine erneute Untersuchung der Elternarten notwendig. Außerdem wurden einige Arten beobachtet, deren Verhalten noch nicht untersucht worden war. Fragestellung und Begründung werden in der Einleitung gegeben.Im zweiten Abschnitt werden einige der vonLorenz gemachten Beobachtungen berichtigt. So zeigten einige der Kreuzungen mitbahamensis, daß die vonLorenz bei eben dieser Art als Kurzhoch-werden bezeichnete Bewegungsweise dem Ab-auf anderer Schwimmenten homolog ist. Ebenso ist die eine der beiden vonLorenz als Kurzhoch-werden bezeichneten Verhaltensweisen des Krickerpels als Ab-auf zu deuten. Der Gruß desflavirostre-Erpels wurde auch bei weiblichen Tieren gesehen. BeiAnas acuta wurde ein Kinnheben festgestellt, das sich in der Form stark vom Kinnheben beiplatyrhynchos unterscheidet. Als neue Verhaltensweisen wurden u. a. das Haltungannehmen und das Tendieren beimflavirostre-Erpel beschrieben.Im dritten Abschnitt werden einige Verhaltensweisen und ihre Funktion diskutiert und der Versuch gemacht, eine Motivationsanalyse zu geben.(Zeichnungen vonHermann Kacher)     

Free Ia Eα chain expression in the E α + :E β − : recombinant strain A.TFR5

Molecular and biochemical techniques have been used to explore the reasons behind low E chain expression in the E ⁺ E ^– I-region recombinant strain, A.TFR5. A.TFR5 (A ^f E ^k, ap5), a recombinant between A.CA (A ^f E ^f) and A.TL (A ^k E ^k), carries the E ^k subregion. Previous results have shown that it expresses the E chain, but at reduced levels relative to E ⁺ E ⁺ strains. No E chains were detected, which is consistent with the A.TFR5E gene being derived from the A.CA parent, which carries the null E ^f allele. In this paper, the defect in E-chain expression is explored. Restriction fragment length polymorphism analysis has localized the recombination event in A.TFR5 approximately 30 kb upstream of E, in the region of the large intervening sequence of E. Northern blot analysis of total RNA from A.TFR5 shows normal amounts of the E message, but no E message. Two-dimensional gel analysis of 15 min pulse-labeled A.TFR5, A.CA, and A.TL E immunoprecipitates shows decreased levels of the intracellular E chain in A.TFR5 relative to A.TL. However, analysis of total cell extracts shows normal levels of this protein. A glycoprotein fraction isolated from total cell extracts of 5 h labeled cells contains normal amounts of intracellular E, but decreased amounts of the mature cell-surface protein. These data suggest that in the absence of E, the E chain (1) takes on an altered conformation that is not as efficiently recognized by alloantibodies, and (2) is found in normal levels as the partially glycosylated intracellular precursor, but is not processed and/or transported efficiently to the cell surface.     

Radial transport of water across cortical sleeves of excised roots ofZea mays L.

The stationary radial volume flows across maize (Zea mays L.) root segments without steles (sleeves) were measured under isobaric conditions. The driving force of the volume flow is an osmotic difference between the internal and external compartment of the root preparations. It is generated by differences in the concentrations of sucrose, raffinose or polyethylene glycol. The flows are linear functions of the corresponding osmotic differences ( ) up to osmotic values which cause plasmolysis. The straight lines obtained pass through the origin. No asymmetry of the osmotic barrier could be detected within the range of driving forces applied ( =±0.5 MPa), corresponding to volume-flow densities of j_{v, s}=±7·10^–8 m·s^–1. Using the literature values for the reflection coefficients of sucrose and polyethylene glycol in intact roots (E. Steudle et al. (1987) Plant Physiol.84, 1220–1234), values for the sleeve hydraulic conductivity of about 1·10^–7 m·s^–1 MPa^–1 were calculated. They are of the same order of magnitude as those reported in the literature for the hydraulic conductivity of intact root segments when hydrostatic pressure is applied.Abbreviations and symbols a _s outer surface of sleeve segment - c concentration of osmotically active solute - j _{v, s} radial volume flow density across sleeve segment - Lp_s hydraulic conductivity of sleeves - Lp_r hydraulic conductivity of intact roots - _N thickness of Nernst diffusion layer - reflection coefficient of root for solute - osmotic value of bulk phase - osmotic coefficient     

Biological chromatology. The laws of colour and design in nature

Summary Biochromatology can be defined as the study of the relationships between colours, and between colours and design in nature. Certain laws concerning biochromatics can be formulated in the terminology of the Colour Circle. One of them is that planes in warm colours and planes in cold colours never adjoin. Another law is that design is never polychromatic. It seems that the observed biochromatical facts can be explained with the help of the principle optimal manifestation of colour, which in turn is based on the formative principles of matter itself.     

Synthetic substrate analogues for UDP-GlcNAc: Manα1-6R β(1-2)-N-acetylglucosaminyltransferase II. Substrate specificity and inhibitors for the enzyme

UDP-GlcNAc: Man1-6R (1-2)-N-acetylglucosaminyltransferase II (GlcNAc-T II; EC 2.4.1.143) is a key enzyme in the synthesis of complexN-glycans. We have tested a series of synthetic analogues of the substrate Man1-6(GlcNAc1-2Man1-3)Man-O-octyl as substrates and inhibitors for rat liver GlcNAc-T II. The enzyme attachesN-acetylglucosamine in 1-2 linkage to the 2-OH of the Man1-6 residue. The 2-deoxy analogue is a competitive inhibitor (K _i=0.13mm). The 2-O-methyl compound does not bind to the enzyme presumably due to steric hindrance. The 3-, 4- and 6-OH groups are not essential for binding or catalysis since the 3-, 4- and 6-deoxy and -O-methyl derivatives are all good substrates. Increasing the size of the substituent at the 3-position to pentyl and substituted pentyl groups causes competitive inhibition (K _i=1.0–2.5mm). We have taken advantage of this effect to synthesize two potentially irreversible GlcNAc-T II inhibitors containing a photolabile 3-O-(4,4-azo)pentyl group and a 3-O-(5-iodoacetamido)pentyl group respectively. The data indicate that none of the hydroxyls of the Man1-6 residue are essential for binding although the 2- and 3-OH face the catalytic site of the enzyme. The 4-OH group of the Man-O-octyl residue is not essential for binding or catalysis since the 4-deoxy derivative is a good substrate; the 4-O-methyl derivative does not bind. This contrasts with GlcNAc-T I which cannot bind to the 4-deoxy-Man- substrate analogue. The data are compatible with our previous observations that a bisectingN-acetylglucosamine at the 4-OH position prevents both GlcNAc-T I and GlcNAc-T II catalysis. However, in the case of GlcNAc-T II, the bisectingN-acetylglucosamine prevents binding due to steric hindrance rather than to removal of an essential OH group. The 3-OH of the Man1-3 is an essential group for GlcNAc-T II since the 3-deoxy derivative does not bind to the enzyme. The trisaccharide GlcNAc1-2Man1-3Man-O-octyl is a good inhibitor (K _i=0.9mm). The above data together with previous studies indicate that binding of the GlcNAc1-2Man1-3Man- arm of the branched substrate to the enzyme is essential for catalysis. Abbreviations: GlcNAc-T I, UDP-GlcNAc:Man1-3R (1-2)-N-acetylglucosaminyltransferase I (EC 2.4.1.101); GlcNAc-T II, UDP-GlcNAc:Man1-6R (1-2)-N-acetylglucosaminyltransferase II (EC 2.4.1.143); MES, 2-(N-morpholino)ethane sulfonic acid monohydrate.     

Techniques for the quantitative study of mutation in plant viruses

Summary Hardly any other virus is chemically and ultramicroscopically as well known as TMV. It is not possible to perform genetic recombinations with this object. The phenomenon of mutation is, however, known and an analysis of the dosis-effect relationship was possible by using the characters chlorotic versus necrotic primary symptoms. Taking into account the phenomenon of interference (mutual exclusion), i.e., comparing the induced mutation frequency with that of a control virus sample diluted to the same level of infectivity, on can perform quantitative analyses. In this way the first chemical mutagensis in the test tube was demonstrated 10 years ago with nitrous acid as mutagenic agent. The criticism raised byBawden to the first publication ofMundry andGierer was already inappropriate at that time. In the meantime it has been demonstrated byWittmann-Liebold andWittmann through analysis of amino acid exchanges in spontaneous mutants and in those isolated after incubation with HNO₂ that the difference between spontaneous and induced mutants demanded byBawden, which cannot be postulated for symptoms in plants, lies, as expected, in amino acid exchanges of the protein coat.

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Zusammenfassung Kaum ein anderes Virus ist chemisch und ultramikroskopisch so gut bekannt wie das TMV. Rekombinations-Genetik ist nicht möglich. Das Phänomen der Mutation ist aber bekannt, und eine Analyse der Dosis-Effekt-Beziehung wurde möglich durch Benutzung der Symptomcharaktere chlorotische versus nekrotische Primärsymptome. Bei Berücksichtigung des Phänomens der Interferenz (mutual exclusion), d. h. wenn man die induzierte Mutationsrate mit der auf gleiche Infektiosität durch Verdünnen der Viruslösung gebrachten als Kontrolle vergleicht, kann eine quantitative Analyse durchgeführt werden. So wurde vor 10 Jahren die erste Chemomutagenese im Reagenzglas mit salpetriger Säure als mutagenes Agens nachgewiesen. Die an der ersten Veröffentlichung vonMundry undGierer vonBawden geäußerte Kritik war schon damals unzutreffend. Inzwischen ist durch die Analyse der Aminosäureaustausche von spontanen und nach Inkubation mit HNO₂ isolierten Mutanten vonWittmann-Liebold undWittmann gezeigt worden, daß die vonBawden geforderte Verschiedenheit spontaner und induzierter Mutanten, die für Symptome an den Pflanzen nicht postuliert werden kann, in den Aminosäureaustauschen des Hüllproteins wie zu erwarten vorhanden ist.

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This paper was a first written for Methods in Virology, Academic Press. The editors and the author did not come to an agreement in the question of citation ofBawden's criticism to the work ofMundry andGierer 1958. It is published here on the occasion of the 10th anniversary of the first chemomutagenesis in the test tube.     

Ist die pflanzliche Photosynthese eine Lichtreaktion Mit rückläufiger Dunkelreaktion? Ein Versuch,Manometrische Licht-Dunkel-Kurven von Chlorella zu Erklären

Zusammenfassung Es wird gezeigt, daß mit Hilfe einer quantitativen O₂-Bestimmungsmethode bei Belichtungsbeginn einer Chlorella-Suspension kein Einquanten-Sauerstoff gefunden wird, wie er aus Manometer-Versuchen vonWarburg erschlossen wurde.Dann wird nachgewiesen, daß die in den Manometergefäßen sich abspielende Zeitreaktion CO₂+H₂OH₂CO₃ den Kreisprozeß der Photosynthese vortäuschen kann, ja sogar vortäuschen muß.Mit 2 Textabbildungen     

Modulation of the serotonin-activated K+ channel by G protein subunits and nucleotides in rat hippocampal neurons

In hippocampal neurons, 5-hydroxytryptamine (5-HT) activates an inwardly rectifying K⁺ current via G protein. We identified the K⁺ channel activated by 5-HT (K_5-HT channel) and studied the effects of G protein subunits and nucleotides on the K⁺ channel kinetics in adult rat hippocampal neurons. In inside-out patches with 10 m 5-HT in the pipette, application of GTP (100 m) to the cytoplasmic side of the membrane activated an inwardly rectifying K⁺ channel with a slope conductance of 36±1 pS (symmetrical 140 mm K⁺) at –60 mV and a mean open time of 1.1±0.1 msec (n=5). Transducin activated the (K_5-HT) channels and this was reversed by -GDP. Whether the K_5-HT channel was activated endogenously (GTP, GTPS) or exogenously (), the presence of 1 mm ATP resulted in a 4-fold increase in channel activity due in large part to the prolongation of the open time duration. These effects of ATP were irreversible and not mimicked by AMPPMP, suggesting that phosphorylation might be involved. However, inhibitors of protein kinases A and C (H-7, staurosporine) and tyrosine kinase (tyrphostin 25) failed to block the effect of ATP. These results show that G activates the G protein-gated K⁺ channel in hippocampal neurons, and that ATP modifies the gating kinetics of the channel, resulting in increased open probability via as yet unknown pathways.     

Direct somatic embryogenesis through thin cell layer culture in Panax ginseng

Direct somatic embryos were differentiated on cotyledon transverse Thin Cell Layers (tTCLs) of Panax ginseng after 9 weeks in the Murashige and Skoog basal (MS) medium containing 2,4-d (5M). When MS medium containing 2,4-d (5M) was used for seedling pretreatment and for tTCLs culture, somatic embryos were observed 2 weeks earlier, i.e. after 7 weeks of culture. On the tTCLs from seedlings pretreated with 2,4-d (5M) combined with benzyladenine and zeatin at 0.1 M (BZ), somatic embryos were observed after 6 weeks of culture and the percentage of embryogenesis was higher (62%) than when 2,4-d was used alone for pretreatment (40%). Similar results were also obtained from pretreatment with combinations of 2,4-d (5M) and thidiazuron (TDZ) (0.01, 0.1M). When a combination of 2,4-d (5M) and BZ (0.1M) was used both for seedling pretreatment and for tTCLs culture, both somatic embryos and shoots were observed after only 3 weeks. As the concentration of BZ increased, the percentage of somatic embryogenesis decreased but the percentage of organogenesis increased. Similar responses were obtained with a combination of 2,4-d (5M) and TDZ (0.01M). On the medium containing both NAA (0.3M) and BZ (1M), globular- and heart- stage embryos developed after 4 weeks of culture into cotyledonary-staged embryos which remained dormant after a short elongation of the embryo axis. The importance of seedling pretreatment by growth substances in enhancing somatic embryogenesis is reported.Abbreviations BA 6-benzyladenine - BZ combination of BA and zeatin - 2,4-d 2,4-dichlorophenoxyacetic acid - MS medium Murashige and Skoog basal medium - NAA a-naphthaleneacetic acid - TDZ thidiazuron - tTCLs transverse thin cell layers - TCL longitudinal thin cell layer