Histochemische Untersuchungen am Auge der Schmeißfliege Calliphora erythrocephala Meig.

Zusammenfassung Im Auge von Calliphora erythrocephala Meig., Mutante chalky, sind zwei ATP-asen, saure und alkalische Phosphatase sowie Glukose-6-Phosphatase histochemisch darstellbar. Die Lokalisation der Enzyme in Retinula und Lamina ganglionaris ist sehr unterschiedlich. 20 min lange Dunkelhaltung der Tiere führt zu Änderungen der Stärke der Nachweisreaktionen einiger Enzyme.

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Histochemical investigations of the eye of the blowfly Calliphora erythrocephala Meig.Part I. ATP-ase, acid phosphatase, alkaline phosphatase, and glucose-6-phosphatase

Summary Within the compound eye of the blow-fly, Calliphora erythrocephala, white-eyed mutant chalky, ATP-ase in two different types, acid and alkaline phosphatase and glucose-6-phosphatase are demonstrated histochemically. In the retina one type of ATP-ase is distributed in the cytoplasm, the other is located at the rhabdomeres. Acid phosphatase activity is located in semper and pigment cells and in the lamina, while alkaline phosphatase is demonstrable only in pigment cells around the crystalline cones and in the lamina. Reaction product of glucose-6-phosphatase is only visible at the basement membrane. Dark adaption about twenty minutes alters the intensity of the reaction of some enzymes.

     

Nachweis einer Pupillenreaktion im Auge der Schmeißfliege

Zusammenfassung Elektronenoptische Untersuchungen mit der Gefrierätzmethode an Augen der Normalform von Calliphora erythrocephala (Meig.) ergeben, daß im dunkeladaptierten Zustand die Rhabdomere 1–6 in den Retinulazellen dicht von Vesikeln umgeben sind, die wahrscheinlich durch Pinocytose an den Mikrovilli der Rhabdomere entstehen. Diese Bläschen fehlen im helladaptierten Zustand nahezu völlig. Die optische Dichte des umgebenden Mediums der lichtleitenden Rhabdomere wird dadurch im Dunkel-Auge herabgesetzt, was mit Hilfe der Becke-Methode im Lichtmikroskop festgestellt werden konnte.Physikalische Berechnungen mit den Gleichungen der geometrischen Optik ergeben, daß durch diese Brechzahländerung in den Sinneszellen im dunkeladaptierten Auge der physiologisch wirksame Lichtfluß in den Rhabdomeren l–6 im grünen Spektralbereich um etwa 30% erhöht werden kann.

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Evidence for a longitudinal pupil in the blowfly eye from studies with light and electron microscopes

Summary Eyes of the wild-type blowfly, Calliphora erythrocephala (Meig.), were investigated by using freeze-etching and the light microscope. In the dark-adapted eye a layer of vesicles 1 m thick borders on the rhabdomeres in sense cells Nos. 1 to 6. These vesicles may arise by pinocytosis at the bases of the rhabdomeric microvilli and decrease the optical density of the sense cell medium adjacent to the rhabdomeres. In the light-adapted sense cells only a few vesicles could be seen. No differences could be observed to exist between the two states of adaptation in the retinular cells Nos. 7 and 8 and in the axial retinular space.Calculations based on geometrical optics show that when rhabdomeres Nos. 1 to 6 are in the dark-adapted state they can transmit about 1.3 times more light energy than when they are in the light-adapted state. The results show the existence of a light-induced pupil reaction correlated to ultrastructural changes in the photoreceptor cells of the blowfly eye.

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Der Deutschen Forschungsgemeinschaft danke ich für die Gewährung eines Ausbildungsstipendiums.     

Direkte Beobachtung der Rhabdomere bei Calliphora erythrocephala (Meig.)

Zusammenfassung Betrachtet man mit einem Mikroskop mittlerer Apertur eine Bildebene im Komplexauge, so kann man bei antidromer Beleuchtung bei Calliphora erythrocephala am intakten Auge innerhalb eines größeren Bereichs gleichzeitig die Rhabdomere eines jeden Ommatidiums deutlich in Form eines virtuellen, aufrechten und vergrößerten Bildes sehen. Die Rhabdomere verschieben sich mit der Zeit distalwärts bei gleichzeitiger Verkleinerung und Wanderung der Pseudopupille. Die Ursachen der Erscheinung bleiben ungeklärt. Es werden Hinweise zur methodischen Ausnützung dieser Erscheinung gegeben; als Beispiel wird die Rhabdomerenanordnung an der Grenze zwischen dem dorsalen und ventralen Augenteil gezeigt.

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Direct observation of the rhabdomeres in Calliphora erythrocephala (Meig.)

Summary If the living eye of Calliphora erythrocephala is illuminated antidromically (from behind) and viewed through a microscope of medium aperture, the rhabdomeres of each ommatidium within a large area can be seen simultaneously at a focal plane within the compound eye. The rhabdomeres appear in a virtual image, upright and magnified. They shift with time towards the cornea and the pseudopupil drifts to the side, becoming smaller. The reasons for this phenomenon remain unclear. Suggestions are made for its use. The effect was employed, for example, to show the pattern of the rhabdomere arrangement at the border between the dorsal and ventral parts of the eye.

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Die Arbeit wurde durch eine Sachbeihilfe der Deutschen Forschungsgemeinschaft an Herrn Prof. Dr. H. Autrum unterstützt.

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Mit Unterstützung des Deutschen Akademischen Austauschdienstes.     

Functional-morphological investigations on the flight muscles and their insertion points in the blowfly Calliphora erythrocephala (Insecta,Diptera)

Summary In flies, for example the blowfly Calliphora erythrocephala, the thorax has fused to form a chitinous capsule. In it we find three functional types of flight muscles, the indirect flight muscles, the direct, and the tension muscles. The indirect or wing beat muscles transfer their power to the capsule which is capable of oscillating. They are suspended nearly horizontally and vertically. The direct muscles used for steering insert laterally on the capsule and go to the wing joint. The third functional type of flight muscle serves to put the lateral walls of the thorax under tension. The site and morphology of the flight muscles are described in detail, making use of 3-dimensional drawings. The flight muscles of Calliphora erythrocephala (Heide 1968) and their functions are compared with those of other dipterans described by different authors.With support of the Deutsche Forschungsgemeinschaft to Professor Nachtigall     

The in vitro production of an anthocyanin from callus cultures of Oxalis linearis

Callus growth and the production of anthocyanins were sustained on the salts and vitamins of Murashige and Skoog. Callus growth was stimulated at a concentration of 8–32 M -naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-d). Benzyladenine (BA) and zeatin at 8 M inhibited callus growth whereas isopentenyladenine (iP) stimulated callus growth. NAA repressed anthocyanin production with an increase in NAA from 8–32 M. Anthocyanin synthesis was promoted by an increase in 2,4-d from 0.5 to 2 M and decreased thereafter up to a concentration 32 M 2,4-d. A concentration of 8 M BA, thidiazuron and zeatin, respectively stimulated pigment production. Sucrose stimulated callus growth at 60 mM and pigment production at 120–360 mM.Abbreviations BA 6-benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - NAA -naphthaleneacetic acid - iP isopentenyladenine - TZ thidiazuron-N-phenyl-N-1,2,3-thiadiazol-5-yl-urea - Bu-HCl Butanol-2N HCl - BAW Butanol-acetic acid-water     

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.     

On head and body movements of flying flies

Head and body movements of flies (Musca domestica and Calliphora erythrocephala) have been studied during sustained flight. Two main points emerge from the analysis: a) Changes in body direction and head direction occur simultaneously in almost all cases. b) During visually guided flight active neck movements are initiated together and in the same direction of body movements. This does not hold in absence of a visual pattern (search). Implications of these findings with respect to the organization of the control system underlying head-body coordination in flies are briefly discussed.     

Polycarnitine—a new biomaterial

The natural product l-carnitine is—due to its biotechnological accessibility and specific properties—on the way to becoming an attractive biobased bulk product. l-Carnitine is a natural betaine with vitamin properties. Carnitine is an essential part of the fatty acid metabolism of human beings and animals. Carnitine was first isolated in 1905 from meat extract and important recent developments include the biosyntheses of l-carnitine from l-lysine or -butyrobetaine. Our synthesis routes are designed to maintain the primary structure and specific properties of carnitine, such as hydrophilicity and stiffening effects for polymeric structures and applications. l-Carnitine is converted via lactonization or olefinization into polymerizable basic molecules. The properties and the applications of carnitine polymers are described.This revised version was published online in February 2005 with text corrections in the subsection Biotechnological production of L-carnitine of the Introduction section.     

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)     

The neurosecretory system of the adult Calliphora erythrocephala

Summary Neurosecretory cells in the brain and suboesophageal ganglion of the adult female Calliphora erythrocephala have been studied in the light microscope. The paraldehydefuchsin stain (PAF) gave by far the clearest pictures.The medial neurosecretory cells of the protocerebrum (m.n.c.) show definite cyclic changes as to the size of the nuclei and the content of secretory material. The cytological changes depend on the age of the fly and the diet given and are correlated with ovarian development.The nuclear size is held to express the metabolic activity of the cells. Cells with large nuclei, as found in young sugar-flies (S1D) and meat-fed flies with developing ovaries (S4D/M2D), contain less secretory material which is released through the axons, while the m.n.c. of old sugar-flies (S6D) have small nuclei and are stuffed with secretory material which is stored in the perikarya.These results confirm those obtained by darkfield microscopy of living m.n.c. by Lea and E.Thomsen (1962 and unpublished).No really convincing evidence for the existence of more than one type of m.n.c. was found.Two small groups of lateral cells were observed. Possibly neurosecretory are further: 1-(2) cells at the base of each optic lobe, two groups of 2–3 cells on the caudal side of the brain, and 2 cells ventrally in the suboesophageal ganglion.Giant neurons of unknown function are situated very near the m.n.c. Their axons join those from the m.n.c., but end in the suboesophageal ganglion.The same region comprises a number of peculiar cells, each containing a large, fluid-filled vacuole (the vacuolated cells). Similar cells are associated with the possibly neurosecretory cells on the caudal side of the brain.My sincere thanks are due to my wife Dr. Ellen Thomsen, who made all the excisions of the brains and the measurements of the nuclei, to T.C.Normann for valuable assistance with the photographic work, and to Mrs. K. Bahnert for technical help with the gallocyanin method. The Carlsberg Foundation has supported the work with grants.     

Zur biologischen Bedeutung des Estrildidengesanges

Zusammenfassung Viele Prachtfinken (Estrildidae) tragen außerhalb der Balz einen ungerichteten Gesang vor. Diese Gesangsform scheint primär mehr oder minder funktionslos und lediglich Ausdruck einer besonders ausgeglichenen Stimmungslage zu sein. Sekundär kann sie infolge großer intraspezifischer Modifikationsbreite ein persönliches Erkennen des 's gewährleisten und durch verschiedenartige Anpassungen (Gruppengesänge, Entwicklung des Zuhörens, Gesangshemmung in Partnernähe mit anschließendem Ablauf intensiver Paarbindungszeremonien) auch die Paar- und Gruppenbindung festigen. Mit dieser Zunahme der sozialen Funktionen wird der Gesang gleichzeitig von der Keimdrüsenreifung immer unabhängiger und ist bei drei der beobachteten Arten (Uraeginthus angolensis, Lagonosticta rhodopareia undAmadina erythrocephala) selbst bei völlig inaktiven Gonaden voll ausgebildet. Auf diese Weise kann er seine sozialen Aufgaben ganzjährig erfüllen.

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Summary Many species of Grass Finches (Estrildidae) sing not only during court-ship (display song) but also on numerous other occasions (solitary song). In general the solitary song does not seem to possess any significant biological function seeming rather to be just a sign of a very tranquil mood, not influenced by any other motivation. In some species, however, it has acquired several new, social functions: Due to its remarkably great individual variability the song of the may contribute to individual recognition within a pair or a breeding colony. It may also promote pair and flock cohesion. In connection with this, several behavioural adaptations have arisen such as singing in duos and trios or peering at the singer by other members of the flock.In theLagonosticta andUraeginthus species males and females possess a song. Solitary song, however, seems to be inhibited by the presence of other birds of the same species, especially the bird's own mate. For this reason the birds are forced to separate from each other for the utterance of solitary song. After having ceased singing they usually come together again and will go through their special greeting ceremonies — the stimulating and synchronizing properties of which are well known — with especially high intensity. Solitary song may thus contribute indirectly to cementing the pair bond.In all species in which solitary song possesses a marked social function it seems to have become more and more independent of gonadal development. Some species (e. g.Uraeginthus angolensis, Lagonosticta rhodopareia, Amadina erythrocephala) sing all the year round and even — as has been proved by histological examination — with completely inactive gonads (stage 1 of the testis cycle, followingBlanchard 1941). It is not clear yet whether song in these species is still sexual or partly social or autochthonous in motivation.The various social functions of solitary song clearly demonstrate thatAndrew's (1961) definition of song seems to be an unnecessarily restricted interpretation which should be abandoned.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

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     

Interactions of γ-L-glutamyltaurine with excitatory aminoacidergic neurotransmission

The in vitro effects of -L-glutamyltaurine on different stages of excitatory aminoacidergic neurotransmission were tested with -D-glutamyltaurine as reference. -L-Glutamyltaurine enhanced the K⁺-stimulated release of [³H]glutamate from cerebral cortical slices (25% at 0.1 mM) and slightly inhibited the uptake by crude brain synaptosomal preparations (about 10% at 1 mM). -L-Glutamyltaurine was also a weak displacer of glutamate and its agonists from their binding sites in brain synaptic membrane preparations, being, however, less selective to quisqualate (QA) sites than -D-glutamyltaurine. The basal influx of Ca²⁺ into cultured cerebellar granular cells was not affected by 1 mM -L-glutamyltaurine, but the glutamate- and its agonist-activated influx was significantly inhibited in low-Mg²⁺ (0.1 mM) and Mg²⁺-free media. The glutamate-evoked increase in free intracellular Ca²⁺ and the kainate-activated formation of cGMP in cerebellar slices were both markedly inhibited by 0.1 mM -L-giutamyltaurine. We propose that -L-glutamyltaurine may act as endogenous modulator in excitatory aminoacidergic neurotransmission.     

Zum Feinbau des Komplexauges von Stylops spec. (Insecta,Strepsiptera)

Zusammenfassung Unter den Cornealinsen des Komplexauges von Stylops befindet sich ein Kristallkegel vom pseudoconen Typ, der von zahlreichen Pigmentzellen umhüllt wird. An seinem proximalen Ende liegen 6 meist pigmentfreie Zellen (Sempersche Zellen).Das Ommatidium besteht aus etwa 60 Retinulazellen. Ihre distal kranzartig miteinander verbundenen Mikrovillisäume bilden ein einziges offenes Rhabdom, das extrazelluläres (?) granuläres Material und die Basis der Semperschen Zellen umgibt. Stellenweise wird das Rhabdom samt granulärem Material von homogen erscheinenden distalen Ausläufern einzelner Retinulazellen überlagert. Proximad zerfällt das Rhabdom zunehmend in kleinere Rhabdomteile. Im zentralen Teil des Ommatidiums liegen 1–2 auffallend große Retinulazellen, die meist weniger elektronendicht erscheinen und kleinere Pigmentgrana haben.Die einzelnen Ommatidien werden von ungemein zahlreichen, sehr pigmentarmen Stützzellen umhüllt. Diese werden — wie die basalen Teile der Retinulazellen — teilweise durch Gliazellfortsätze isoliert.Bei Stylops, einem Vertreter der Strepsipteren, handelt es sich nicht um ocelläre Komplexaugen (Strohm, 1910), auch nicht um eucone Ommatidien (Kinzelbach, 1967), sondern um Ommatidien vom pseudoconen Typ. Zumindest der Bau des Rhabdoms ähnelt dem des Larvenauges (Stemma), dessen rezeptorischer Teil entgegen den Annahmen früherer Autoren in der Imago nicht reduziert wird.

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On the fine structure of the compound eye of Stylops spec. (Insecta, Strepsiptera)

Summary In the compound eye of Stylops a crystalline cone of the pseudocone type is found beneath the corneal lens. It is enveloped by several pigment cells. At the proximal part of the cone there are 6 cells (Semper cells) mostly pigment-free.The ommatidium consists of approximately 60 retinula cells. Their rhabdomeres distally rim-like connected to another form a single open rhabdom which encircles extracellular granular material as well as the bases of the Semper cells. Here and there the rhabdom plus granular material is overlain with distal protrusions of single retinula cells which appear to be homogeneous. Towards the proximal part the rhabdom increasingly divides up into smaller rhabdomal segments. One or two conspicuous large retinula cells were found in the central part of the ommatidium, appearing to be less electron-dense and containing pigment granules of a smaller size. Each ommatidium is surrounded by numerous cells (Stützzellen) lacking in pigment. These cells are partially insulated from another—as well as the basal parts of retinula cells—by protrusions of glia cells.Our investigations show that the eyes of Stylops (as a representative of Strepsiptera) are not of the ocellar complex eye type. At least the structure of the rhabdom resembles to that of the larval eye (stemma), the receptor part of which is not reduced in the imago.

Herrn Prof. Dr. Helmcke danke ich für die freundliche Unterstützung am Raster-Elektronenmikroskop.     

Organisation of the compound eye of a tipulid fly during the day and night

Summary The ultrastructure of the compound eye of the Australian tipulid fly,Ptilogyna spectabilis, is described. The ommatidia are of the acone type. The rhabdom corresponds to the basic dipteran pattern with six outer rhabdomeres from retinular cells 1–6 (R1-6) that surround two tiered central rhabdomeres from R7 and 8. Distally, for about 8 m, the rhabdom is closed. For the remainder, where the rhabdomere of R8 replaces that of R7, the rhabdom is open, and the rhabdomeres lie in a large central ommatidial extracellular space. In the proximal two thirds of the rhabdom, the central space is partitioned by processes from the retinular cells so that the individual rhabdomeres are contained in pockets.At night the rhabdom abuts the cone cells, but during the day it migrates some 20 m proximally and is connected to a narrow (1–2 m) cone cell tract. This tract is surrounded by two primary pigment cells, which occupy a more lateral position at night and thus act like an iris. Pigment in secondary pigment cells also migrates so as to screen orthodromic light above the rhabdom during the day. Between midday and midnight, the rhabdom changes in length and cross-sectional area as a result of asynchrony of the shedding and synthetic phases of photoreceptor membrane turnover. The effects of these daily adaptive changes on photon capture ability are discussed with regard to the sensitivity of the eye.     

Über die biologie blutsaugender simuliiden (diptera)

Summary Swarming behaviour of the mammalophilic simuliids Wilhelmia equina, W. salopiensis and Boophthora erythrocephala is closely related to sex-finding. Long-range orientation occurs by optically directed swarming of the towards a dark marker above them contrasting with the bright sky. flying directly to the same marker or the bigger object carrying it, e.g. a tree, are visually recognized by the at a short distance (short-range orientation).Sex-orientation shows the following pattern of behaviour: Weather conditions (temperature, relative humidity, covering of the skyfree, partially or covered) may vary to a great extent without obvious effect. If brightness of the sky exceeds 5000 lux and velocity of the wind is limited to 10 m/sec measured 1 m above the ground, and begin to seek for moving (e.g. blood hosts of , adequate dummies) or motionless conspicuous objects (e.g. trees, cottages).Around these objects behaviour differs between species, but more markedly between the sexes: are flying directly to the object and sit down, are swarming in a short distance from it, waiting for the arrival of more . As the emerge from the pupae earlier than the which are not yet inseminated, they are more numerous. Furthermore, a are swarming longer than one day (demonstrated by tagging experiments) so that every may copulate several times.Orientation of towards the optic marker is the last stage in the long-range orientation. With regard to of W. equina, the pattern is as follows: The occupying the uppermost flying position in the whole swarm orientates itself in such a manner that the image of the marker is centrally in the median space of its head, which is covered by both complex eyes having an overlapping space of 38°. Even during individual movements this position is maintained so that the marker leaves this space only partially. Moreover the remains at such a distance that the image of the marker neither exceeds the anatomical field of vision of the dorsal parts of the complex eyes in the sagittal plane (54°) nor disappears from it completely. Accordingly, the vertical distance from the marker depends on its size, but at a maximum distance of 60 cms the fly changes its orientation, using a portion of the marker, such as a corner, as new fixpoint. Orientation to the marker is not restricted to the background of the open sky; other backgrounds of the same colour are accepted, too, if they are smaller than the anatomical field of vision in the median sagittal plane (54°). Furthermore, lateral parts of the same field of vision, which is 158° in the transversal plane, may be covered up to one half of its width by dark objects, whereas no object is tolerated touching the median sagittal field of vision in front. This demonstrates, too, that not visual perception as such, but the special behaviour of the simuliids is related to the anatomical field of vision.With W. salopiensis, W. equina, and B. erythrocephala meeting and recognizing of the partner for copulation usually occurs during flight. Both partners, however, fall down to the ground immediately thereafter and seperate within 3–5 seconds. In the final stage of copulation the partners are in opposite position, but the abdomina remain in normal position, i.e. turned at 180°, the , lying on its back with its abdomen not twisted. Compared with the copulation among Culicidae and Ceratopogonidae, this position must be considered more primitive.The special differentiation of the complex eyes of male simuliids permits them to recognize both distances (as indicated by the orientation to the marker) and moving objects, enabling them to distinguish between flying , which will be pursued, and in the same swarm, the presence of which causes no reaction, and enemies such as hovering flies and preying hymenoptera, the arrival of which produces a flight reaction. All these actions are realized by the coarsely screened dorsal parts of the complex eyes.The anatomical position of the male genitalia must be considered morphologically primitive, as the hypopygium lacks inversion. Transfer of sperms is by means of a spermatophore. In the female, the accessory glands of the 8th segment open into the ductus receptaculi, whereas the glands of the 9th segment open to the outside close to the gonoporus.

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Herrn Oberlehrer i. R. G. Schlörer zum 75. Geburtstag gewidmet.     

Methylobacterium rhodesianum MB 126 possesses two acetoacetyl-CoA reductases

Two constitutive acetoacetyl-CoA (AcAc-CoA) reductases were purified from Methylobacterium rhodesianum MB 126, an NADPH-linked d(-)--hydroxybutyryl-CoA forming reductase (enzyme A) and an NADH-and NADPH-linked l(+)--hydroxybutyryl-CoA forming reductase (enzyme B). Enzyme A and B give apparent K _m values of 15 M and 30 M for AcAc-CoA, 18 M for NADPH and 30 M for NADH, respectively. They are inhibited by AcAc-CoA at concentrations higher than 25 M and 50 M, respectively. The contribution of the two reductases to poly--hydroxybutyrate synthesis is discussed.     

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     

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.     

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