Non-coordinate synthesis of RNA polymerase beta beta'' subunits in a temperature-sensitive beta''-subunit mutant of Escherichia coli

Summary On exposure to high temperature of a temperature-sensitive RNA polymerase subunit (rpoC92) mutant of Escherichia coli, selective reduction was observed in the rate of synthesis of a group of proteins including RNA polymerase subunit. The finding that the synthesis of subunit but not subunit was specifically repressed in this mutant grown at non-permissive temperature indicates that the functionally intact RNA polymerase is required for the synthesis of subunits be coordinated. In addition, the assembly of newly synthesized RNA polymerase subunits was inefficient in this mutant at the steps where altered subunit was involved, and the unassembled enzyme subunits were rapidly and preferentially degraded. During recovery to non-restricted growth, the synthesis of both and subunits was transiently enhanced in parallel leading to recovery of the intracellular concentration of functional RNA polymerase.     

Variability in the 11S globulin fraction of seed storage proteins ofHelianthus (Asteraceae)

The seed storage globulins from sixHelianthus and four hybrids were studied using mono and bidimensional gel SDS electrophoresis (+ 2 mercaptoethanol). The polypeptide composition of each subunit was determined. Different pairs are specifically expressed according to the species studied. Three typical patterns were discriminated. All the studied species exhibit five subunits: two of them are expressed in all the species (₁₁ and ₂₂). The subunit corresponding to the ₁₁ pair is present inH. petiolaris and in the three populations ofH. annuus studied. The _2b2 pair is common toH. annuus andH. argophyllus. H. petiolaris presents two specific _2a2 and ₄₄ pairs andH. annuus a specific ₃₃ pair. InH. argophyllus _{11 33} or ₄₄ are never observed but are replaced by ₁₃ and ₃₁ pairs. Some globulins, poorly represented, are of forms but present chains of higher molecular weights (in the range 54–56 kDa). Expressing variations in the banding patterns between these species by the use of a similarity index reveals complete identity between the three populations ofH. annuus. Identity between the twoH. petiolaris studied is also observed.H. annuus andH. argophyllus appear to be closer to each other thanH. petiolaris concerning the seed storage globulins.     

The relation of proton motive force,adenylate energy charge and phosphorylation potential to the specific growth rate and efficiency of energy transduction inBacillus licheniformis under aerobic growth conditions

The magnitude of the proton motive force (p) and its constituents, the electrical () and chemical potential (-ZpH), were established for chemostat cultures of a protease-producing, relaxed (rel ^–) variant and a not protease-producing, stringent (rel ⁺) variant of an industrial strain ofBacillus licheniformis (respectively referred to as the A- and the B-type). For both types, an inverse relation of p with the specific growth rate was found. The calculated intracellular pH (pH_in) was not constant but inversely related to . This change in pH_in might be related to regulatory functions of metabolism but a regulatory role for pH_in itself could not be envisaged. Measurement of the adenylate energy charge (EC) showed a direct relation with for glucose-limited chemostat cultures; in nitrogen-limited chemostat cultures, the EC showed an approximately constant value at low and an increased value at higher . For both limitations, the ATP/ADP ratio was directly related to .The phosphorylation potential (G'p) was invariant with . From the values for G'p and p, a variable H⁺/ATP-stoichiometry was inferred: H⁺/ATP=1.83+0.52µ, so that at a given H⁺/O-ratio of four (4), the apparent P/O-ratio (inferred from regression analysis) showed a decline of 2.16 to 1.87 for =0 to _max (we discuss how more than half of this decline will be independent of any change in internal cell-volume). We propose that the constancy of G'p and the decrease in the efficiency of energy-conservation (P/O-value) with increasing are a way in which the cells try to cope with an apparent less than perfect coordination between anabolism and catabolism to keep up the highest possible with a minimum loss of growth-efficiency. Protease production in nitrogen-limited cultures as compared to glucose-limited cultures, and the difference between the A- and B-type, could not be explained by a different energy-status of the cells.Abbreviations CCCP carbonylcyanide-p-trichloromethoxyphenylhydrazone - DW dry weight of biomass - F Faraday's constant, 96.6 J/(mV × mol) - Fo chemostat outflow-rate (ml/h) - FCCP carbonylcyanide-p-trifluoromethoxyphenylhydrazone - G'p phosphorylation potential, the Gibbs energy change for ATP-synthesis from ADP and P_i - G'⁰p standard Gibbs energy change at specified conditions - H⁺/ATP number of protons translocated through - ATP synthase in synthesis of one ATP - H⁺/O protons translocated during transfer of 2 electrons from substrate to oxygen - specific growth rate (1/h) - _H+ transmembrane electrochemical proton potential, J/mol - M_b molar weight (147.6 g/mol) of bacteria with general cell formula C_6.0H_10.8O_3.0N_1.2 - pH_out,in extracellular, intracellular pH - P_i (intracellular) inorganic phosphate - p proton motive force, mV - pH transmembrane pH-difference - transmembrane electrical potential, mV - P/O number of ADP phosphorylated to ATP upon reduction of one O^2– to H₂O by two electrons transferred through the electron transfer chain - P/O (H⁺/O) × (H⁺/ATP)^–1 - P/O_F, P/O_N P/O with the two electrons donated by resp. (NADH + H⁺) and FADH - q specific rate of consumption or production (mol/g DW × h) - rel ⁺,rel ^– stringent, relaxed genotype - R universal gas constant, 8.36 J/(mol × degree) - T absolute temperature - TPMP⁺ triphenylmethylphosphonium ion - TPP⁺ tetraphenyl phosphonium ion - Y growth yield, g DW/mol - Z conversion constant=61.8 mV for 310 K (37 °C) - ZpH transmembrane proton potential or chemical potential, mV     

Die ?Stresemannsche Revolution“ in der Ornithologie des frühen 20. Jahrhunderts

Zusammenfassung Erwin Stresemann (1889–1972), Generalsekretär, Präsident und Ehrenpräsident der Deutschen Ornithologischen Gesellschaft (bzw. Deutschen Ornithologen-Gesellschaft) über 50 Jahre, war einer der hervorragendsten Ornithologen des 20. Jahrhunderts. In den 1920er und 1930er Jahren gab er den Anstoß für eine weltweite Transformation der älteren, vorwiegend systematisch-faunistischen Ornithologie zu einem Zweig der modernen Biologie und beeinflusste einen großen Kreis von Zeitgenossen (die Stresemannsche Revolution). Mit seinem maßgeblichen Aves-Band (1927–1934) des Handbuchs der Zoologie und den Dissertationen seiner Schüler entstand durch Verbindungen mit der Genetik, der funktionellen Morphologie, der Physiologie und Ethologie der Vögel eine Neue Biologische Ornithologie.

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The Stresemann Revolution in ornithology during the early 20^th century

Summary Erwin Stresemann (1889–1972), Secretary General, President and Honorary President of the Society of German Ornithologists for 50 years, was one of the outstanding ornithologists of the 20^th century. During the 1920s and 1930s, he initiated the global transformation of the traditional ornithology, which had been primarily systematic and faunistic in scope, into a branch of modern biological science, a New Avian Biology, and influenced a large circle of contemporaries (the Stresemann Revolution). He forged links, directly or indirectly, between ornithology and genetics, functional morphology, physiology and ethology, when he published his seminal volume Aves (1927–1934) in the German Handbook of Zoology, and instigated the theses of a large number of PhD students.

     

In vitro propagation of Asparagus maritimus – A rare Mediterranean salt-resistant species

Asparagus maritimus L. Miller is a rare species growing of the Mediterranean region and is morphologically similar to A. officinalis. In order to establish an efficient in vitro propagation protocol, explants were excised from spear segments and cultured on Murashige and Skoog (1962) medium containing 3% sucrose and various concentrations of growth regulators. The best shoot initiation (3–4 per explant) was achieved on a medium containing 0.88 M N⁶-benzyladenine (BA), 0.93 M kinetin, 1.07 M -naphthaleneacetic acid (NAA) and 3.90 M ancymidol. Shoot initiation could also be achieved without ancymidol but the shoots were thinner and longer. A very high shoot multiplication rate was achieved on media supplemented with 3% sucrose, 1.07 M NAA, 0.93 M kinetin, 0.44 M BA and various concentrations of ancymidol. The lowest concentration of ancymidol (0.39 M) significantly promoted the highest shoot multiplication rate (11.9 shoots/crown). For root formation, media were supplemented with 6% sucrose, 1.07 M NAA and various concentrations of ancymidol. Rooting frequency increased with higher ancymidol concentration up to 5.07 M (82.0% rooting). The number of ex vitro shoots formed was strongly correlated (r=0.66) with the length of roots formed in vitro, which was the highest at a 1.95 M ancymidol.     

Synthesis of hydroxycinnamic acid esters of betacyanins via 1-O-acylglucosides of hydroxycinnamic acids by protein preparations from cell suspension cultures of Chenopodium rubrum and petals of Lampranthus sociorum

Protein preparations from cell suspension cultures of Chenopodium rubrum L. and petals of Lampranthus sociorum (L.Bol.) N.E.Br. (Mes.C.L.Bol.) catalyzed the formation of acylated betacyanins, i.e. celosianin I and II (p-coumaroyl-and feruloylamaranthins) and lampranthin I and II (p-coumaroyl- and feruloylbetanins), from 1-O-(p-coumaroyl)-and 1-O-feruloyl--glucoses as acyldonors and the respective acceptor molecules amaranthin (betanidin 5-O-sophorobiuronic acid = betanidin 5-O--[12]-glucuronosyl--glucoside) and betanin (betanidin 5-O--glucoside). The enzymes involved could generally be classified as 1-O-hydroxycinnamoyl--glucose:betanidinglycoside O-hydroxycinnamoyltransferases (EC 2.3.1.-).Abbreviations HCA hydroxycinnamic acid - HCA hydroxycinnamoyl (=hydroxycinnamic acid-ester moiety) - HPLC high-performance liquid chromatography - TLC thin-layer chromatography     

The glycolipid specificity ofErythrina cristagalli agglutinin

The interaction of¹²⁵I-labeledErythrina cristagalli agglutinin (ECA) with neutral glycosphingolipids on thin layer chromatograms was examined by the overlay technique followed by radioautography. The lectin bound topara-globoside with a sensitivity about 10 times higher than to lactosylceramide or globoside, in agreement with the specificity of the lectin forN-acetyllactosamine. The lower limit of detection ofpara-globoside was about 0.66 nmol. The specific binding of ECA to this glycolipid was confirmed by a highly sensitive enzyme-linked lectin assay (ELLA), utilizing the horseradish peroxidase-avidin-biotin system for detection of bound lectin. Overlays of neutral glycosphingolipid extracts from human erythrocyte membranes and from human granulocytes with ECA demonstrated that the lectin can be employed for the detection of small amounts ofpara-globoside in biological materials also in the presence of excess globoside. No staining was obtained when thin layer chromatograms of neutral glycosphingolipid extracts from rabbit erythrocyte membranes were overlayed with¹²⁵I-ECA. Afterin situ treatment of the chromatograms with -galactosidase, the lectin bound to several components, one of which had a mobility corresponding to that of the pentahexosylceramide Gal3Gal4GlcNAc3Gal4Glc1Cer, the major neutral glycosphingolipid of rabbit erythrocytes, thus providing further evidence for the specificity of ECA forpara-globoside.Abbreviations GSL glycosphingolipid(s) - CDH lactosylceramide, Gal4Glc1Cer - CTH trihexosylceramide, Gal4Gal4Glc1Cer - GLOB globoside, GalNac3Gal4Gal4Glc1Cer - PG para-globoside, Gal4GlcNAc3Gal4Glc1Cer - AsG_M1 asialo-G_M1, Gal3GalNAc4Gal4Glc1Cer - FORS Forsmann antigen, GalNAc3GalNAc3Gal4Gal4Glc1Cer - CPH pentahexosylceramide, Gal3Gal4GlcNAc3Gal4Glc1Cer - ECA Erythrina cristagalli agglutinin - SBA soybean agglutinin - PBS phosphate-buffered saline - PVP-40 polyvinylpyrrolidone M.W. 40000 - BSA bovine serum albumin - HRP-avidin horseradish peroxidase conjugated to avidin - ELLA enzyme-linked lectin assay - ELISA enzyme-linked immunosorbent assay - PMNL polymorphonuclear leukocytes - HPTLC high performance thin layer chromatography     

Existence and uniqueness of a sharp travelling wave in degenerate non-linear diffusion Fisher-KPP equations

In this paper we use a dynamical systems approach to prove the existence of a unique critical value c ^* of the speed c for which the degenerate density-dependent diffusion equation u _ct = [D(u)u _x ] _x + g(u) has: 1. no travelling wave solutions for 0 < c < c ^*, 2. a travelling wave solution u(x, t) = (x - c ^* t) of sharp type satisfying (– ) = 1, () = 0 ^*; '(^*–) = – c ^*/D'(0), '(^*+) = 0 and 3. a continuum of travelling wave solutions of monotone decreasing front type for each c > c ^*. These fronts satisfy the boundary conditions (– ) = 1, '(– ) = (+ ) = '(+ ) = 0. We illustrate our analytical results with some numerical solutions.     

Aufbau, Variabilit?t und m?gliche Funktionen des Rufduetts beim ThermometerhuhnLeipoa ocellata

Zusammenfassung Beim ThermometerhuhnLeipoa ocellata tragen die Partner eine Paares ein Rufduett vor. Der Anteil des besteht aus einer Rufreihe, die sich aus einer Folge von 2–7 identischen, zweisilbigen Rufen zusammensetzt. Das trägt einen einzelnen, obertonreichen und langgezogenen Ruf vor. Sowohl der Ruf des als auch die Rufreihe des wird in Serien vorgetragen. Innerhalb einer solcher Ruf- bzw. Rufreihenserie können mehrere Duette auftreten. Die Rufe sind jedoch nicht ausschließlich an das Duett gebunden. Die Variabilität im Aufbau des Duetts äußert sich im Zeitpunkt des Einsatzes des antwortenden Vogels, in der Anzahl der -Rufe während des Duetts und in der Anzahl der Einheiten, aus denen sich der Duettanteil des zusammensetzt. Das beginnt signifikant häufiger als das eine Serie, in der ein oder mehrere Duette vorkommen. Ebenso ist es häufiger der Initiator des ersten in dieser Serie liegenden Duetts. Das Duett dient wohl hauptsächlich zur Festigung des Zusammenhalts zwischen den Paarpartnern. Es erfüllt jedoch von seinen physikalischen Eigenschaften her auch die Bedingungen, die für ein territorial wirksames Signal gelten.

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Structure, variability and possible functions of duetting in the Mallee FowlLeipoa ocellata

Summary In the Australian Mallee Fowl,Leipoa ocellata, both and of a pair are involved in a call duet. The part of the consists of a sequence of 2–7 identical two-syllable calls. The contributes a single long-drawn-out call rich in harmonics. The call of the as well as the call sequence of the are presented in series. Within a series of calls () or call sequences () several duets can occur. The respective vocalizations, however, do not exclusively occur during the duet.The variability in the details of the duet expresses itself in the lag period after which the mate responds, in the number of -calls during the duet, and in the number of calls within the call sequence of the . The begins a series during which one or several duets occur significantly more frequently than the . The circumstances under which duetting occurs indicate that duet calling mainly serves to maintain the pair bond. Moreover, due to its physical characteristics the duet also seems to be suited to serve as a territorial signal.

     

A phylogenetic analysis ofLeucaena (Leguminosae: Mimosoideae)

Chloroplast DNA restriction fragment length polymorphisms have been used to reconstruct the maternal phylogeny of all the known taxa in the small neotropical legume genusLeucaena. Three major plastome clades were recognized, but these did not conform with relationships between the taxa proposed on other characters from morphology, cytology or hybridization. The maternal parentage of tetraploids within the genus has been proposed. Evidence for introgression was found between diploidL. diversifolia and tetraploidL. diversifolia. The implications of these results for the origin of the cultivated taxa are discussed.     

Genotypic control of pollen plant formation in Nicotiana tabacum L.

Summary Tobacco plants (Nicotiana tabacum L.) of four varieties (Badischer Burley, White Burley, Techne, Kupchunos) were raised at different temperatures and daylengths and the effect of genotype on embryogenic pollen grain formation in situ and on pollen plant formation in anther and pollen cultures from these plants was studied. Genotype controlled embryogenic pollen grain and pollen plant formation by defining productivity under standard growth conditions (long days at 24 °C). Kupchunos was the most productive variety, followed by White Burley, Techne, and Badischer Burley. Furthermore, genotype defined which environmental factor was able to affect embryogenic pollen grain and pollen plant formation and also to which degree. In anther cultures, in addition to these effects, genotype controlled the formation of (an) inhibitory substance(s) in the anther wall in interaction with the plant growth conditions. In Badischer Burley and Techne, inhibitor action could be prevented by isolation of the pollen after one week of anther culture. Finally, direct pollen cultures in Badischer Burley and Techne produced embryos were only when the pollen was isolated from nearly mature anthers, while in White Burley and Kupchunos, embryos also produced at earlier stages and at higher yields. This indicated that genotype controls the time when the embryogenic pollen grains become ready to divide. The results are discussed in relation to strategies to overcome recalcitrance of species and genotypes.     

Production of a novel syrup containing neofructo-oligosaccharides by the cells of Penicillium citrinum

A novel syrup containing neofructo-oligosaccharides was produced from sucrose (Brix 70) by whole cells of Penicillium citrinum. The efficiency of fructo-oligosaccharides production was more than 55% and those of the main carbohydrate components, 1-kestose (Fruf 21Fruf 21 Glc), nystose (Fruf 21Fruf 21 Fruf 21 Glc) and neokestose (Fruf 26 Glc12 Fruf), were 22, 14 and 11%, respectively.     

Zur Paarbildung der Uferschwalbe (Riparia riparia)

Zusammenfassung Uferschwalben kehren aus den afrikanischen Winterquartieren in Trupps beiderlei Geschlechts zurück. Erste Beringungsergebnisse belegen, daß zunächst mehrjährige, vermutlich untereinander bekannte Vögel eintreffen, die den Brutplatz aus vergangenen Brutperioden her kennen. Die Masse der später ankommenden Vögel dürfte weitgehend aus einjährigen oder ortsfremden Uferschwalben bestehen, die sich größtenteils erst während der Paarbildung persönlich kennenlernen. Der anfängliche Schwarmzusammenhalt der nacheinander eintreffenden Trupps führt zur Bildung von Subkolonien, die für Brutplätze ab einer bestimmten Größenordnung typisch sind. Uferschwalben- gründen nacheinander mehrere Reviere, d. h. sie besetzen Steilwandbereiche, in denen sie ausschließlich mit den Füßen eine Röhre oder Mulde graben, singen und Bogenflüge starten. Bis auf singende oder bekannte werden Artgenossen im Revier geduldet. Uferschwalben- suchen besetzte Reviere auf. Ohne Röhrenbindung verhalten sie sich still und unauffällig, ihre Grabungsaktivitäten sind von untergeordneter Bedeutung. Die Bindung an ein bestimmtes Revier entwickelt sich individuell verschieden und entscheidet über den Abschluß des Röhrenbaues (Herstellung der Nistkammer). Reviere ohne dauerhafte -Bindung werden von den aufgegeben. Aktivitäten, die auf wachsende Revierbindung eines hindeuten, sind: häufige oder/und länger dauernde Aufenthalte des in einem besetzten Revier und sporadisches Mitgraben; aggressives Verhalten gegenüber Artgenossen (i. d. R. fremde ), die im Revier landen wollen; gemeinschaftlicher, leiser Gesang von und im Röhrenbereich. Aktivitäten, die für eine vollzogene Paarbildung sprechen, sind: Fertigstellen der Röhre durch Grabung der Nestkammer; längere gemeinsame Aufenthalte innerhalb und außerhalb der Röhre; Voranfliegen des beim Röhrenanflug; Übernachten von und in der Röhre; Nestbau; ausdauernde Verfolgungsflüge während der Kopulationsphase. Die Paarbildung ist demnach ein individueller Prozeß, bei dem die Aktivitäten der im Revier als Werbung, die der als Revierwahl interpretiert werden.

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On pair-formation in the Sand Martin,Riparia riparia

Summary European Sand Martins arrive at their breeding sites in flocks of usually unmated and . Ringing results of a large population in NW-Germany and own observations indicate that the first flocks about a dozen individuals with an approximately balanced sex ratio appear at traditional breeding places and consist of older, experienced resident birds (presumably acquainted with one another). The birds arriving over the next several weeks are mainly first-year or non-resident individuals. The flocks arrive separately in areas with suitable sandcliffs, synchronize the pair-formation activities and avoid disturbances among paired and unpaired birds. This behaviour causes the formation of subcolonies, which are typical for all densely occupied breeding places. Each settles on a fixed area on the sandcliff (territory) in order to excavate a burrow, to sing the territory-song (fig. 2 b) and to perfor the territory-circle-flight (fig. 2 c, 4 a). Silent birds (normally ) are welcomed or tolerated by the resident . The sexes are monomorphic and therefore courtship displays of the are non-aggressive until establishment of pair-bonds. Only intruding singing or individually known neighbouring are driven away, usually at early stages of territory occupation. Unmated are normally shy and very sensitive to protracted disturbances. visit several occupied territories of the colony (fig. 1–3) in order to choose a burrow. leave territories which do not attract a . They settle new territories on the sandcliff, causing a surplus of burrows compared to breeding pairs in the colony. Activities which indicate the development of pair-bonds are: regular visits of a to a particular occupied territory with sporadic excavations by the ; aggressive activities of the towards other visitors usually , but sometimes at first even against the resident (i. e.: vocal threats, bill-gaping, pecking or pushing with the bill or vigorous face-to-face fights, fig. 3 b, 3 c). and sing the soft mating song at or in the burrow (fig. 1 c). Activities which indicate completed pair-bonds are: completion of the burrow by digging the nestchamber, predominantly done by the ; both birds staying together over long periods, both inside and outside the burrow; invitation-flight by the (fig. 4 b); and spending the night together in the burrow; beginning of nest-building, first only by the , then by both birds and finally only by the , accompanied by the (guarding-flight;, fig. 4c); mate-pursuit flights (sexual chases) during copulation phase, in which the singing pursues the silent , often accompanied by other (cp. fig. 4 d). Pair-formation in the Sand Martin occurs on individual territories and not, according toHickling (1959), within the flock.

     

Development of high frequency multiple shoot formation in Persian clover (<Emphasis Type="Italic">Trifolium resupinatum</Emphasis> L.)

An efficient and reliable micropropagation system for Persian clover (Trifolium resupinatum L.) was developed using different explants and media. Node, hypocotyl and cotyledonary node explants were cultured on Murashige and Skoog (MS) medium supplemented with combinations of either 6-benzyladenine (BA) and indole-3-butyric acid (IBA) or BA, Kinetin (KIN) and IBA. Direct multiple shoots developed within 6weeks in all explants in most media tested. The best shoot multiplication capacity was obtained from cotyledonary node explants on MS medium containing 7.1M BA and 1M IBA or 14.1M BA and 1M IBA. Elongated shoots were rooted on either MS medium alone or combination with different concentrations of indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) and -naphthaleneacetic acid (NAA). High rooting was achieved in half strength MS medium containing 8M IBA.     

Comparison of human and chimpanzee ξ1 blobin genes

Summary The DNA base sequences of the entire chimpanzee 1 globin gene and an additional 1 kb of DNA flanking both the human and chimpanzee genes have been determined. Whereas the human 1 gene contains a termination codon in the sixth position, the chimpanzee gene appears to be functional. This finding confirms Proudfoot et al.'s suggestion that the human 1 gene was recently inactivated. Like the corresponding human 1 and 2 genes, the first and second introns of the chimpanzee 1 gene are occupied largely by tandem repeats of short oligonucleotides. These tandem repeats have undergone several rearrangements since the divergence of the human and chimpanzee 1 genes.     

Characterization of solute/proton cotransport in plasma membrane vesicles from Ricinus cotyledons,and a comparison with other tissues

Plasma membrane vesicles, purified by aqueous two-phase partitioning, were used to investigate the presence of sugar and amino acid carriers in cotyledons and roots of Ricinus communis L. and in roots of red beet (Beta vulgaris L.). Artificial pH and electrical gradients were generated across the plasma membrane, and [¹⁴C]acetate and [¹⁴C]tetraphenylphosphonium were used to demonstrate the presence of an internal alkaline pH gradient and an internal negative membrane potential, respectively. In Ricinus cotyledons, uptake of sucrose was more strongly inhibited than that of glutamine by p-chloromercuribenzenesulphonic acid, phlorizin and phenylglyoxal. The sucrose transport system showed a high degree of substrate specificity with only the presence of maltose and phenyl--glucoside significantly affecting sucrose uptake; in contrast, the glutamine transport system was inhibited by a number of other amino acids. pH+gD-driven glutamine uptake showed saturation kinetics with a K _m of 0.35 mol · m^–3. Sucrose and glutamine -driven uptake was pH dependent with an optimum in the acidic range (pH 6.25) and a decrease at higher pH values. Vesicles obtained from cotyledons and roots of Ricinus showed different transport properties. In the cotyledons, gDH+gD-driven transport for both sucrose and glutamine were observed at similar levels; however, in the root tissue, pH--driven glutamine transport was the dominant uptake process. Uptake rates for glucose and fructose were low in the cotyledons whereas, in the roots, glucose and sucrose transport were slightly higher than that of fructose. In vesicles from red beet tissue there was a different uptake profile, with evidence of proton-coupled cotransport systems for sucrose and glucose, but lower uptake of glutamine and fructose. The results are discussed in relation to the reported different pathways for loading and unloading of solutes in these tissues.Abbreviations CCCP carbonyl cyanide-m-chlorophyenyl hydrazone - DEPC diethyl pyrocarbonate - NEM N-ethylmaleimide - PCMBS p-chloromercuribenzenesulfonic acid - TPP tetraphenylphosphonium ion - gDH⁺ proton electrochemical potential gradient - membrane potential We would like to thank the SERC(UK) and the Royal Society for financial support.     

Visualization of the uptake of high-density lipoprotein by rat aortic endothelial cells and smooth muscle cells in vitro

High-density lipoproteins (HDL) were conjugated to Fluorescein 1,1-dioctadecyl 3,3,3,3-tetramethylindocarbocyanine perchlorate (DiI) or colloidal gold for the investigation of ultrastructural aspects of binding and uptake of HDL by cholesterol-loaded cultured endothelial and smooth muscle cells from rat aorta. When cells were incubated for 2h at 4°C, HDL–DiI and HDL–gold conjugates were seen only on the cell surface. When cells were returned to incubation at 37°C for 5min, HDL–DiI appeared in the cytoplasm and colocalized with the fluorescent cholesteryl ester tag BODIPY-FL-C₁₂. HDL–gold conjugates appeared in the plasmalemmal invaginations and plasmalemmal vesicles. After incubation for 15min, most of the HDL–gold conjugates reappeared on the cell surface. After incubation for 30min, only a few conjugates were observed and they localized in lysosomal-like bodies. Quantitative data indicated that when the cholesterol-loaded cells were incubated at 4°C for 2h, the numbers of HDL–gold associated in clusters on the endothelial cell surface was 1.18 clusters/m. When cells were returned to incubation at 37°C for 5min, this value decreased to 0.7, increased again to 1.13 at 15min, and decreased to 0.29 at 30min. The numbers of clusters in the plasmalemmal invaginations were 0.06 clusters/m at 4°C for 2h, increased to 0.34 at 37°C for 5min and decreased gradually to 0.19 and 0.04 at 15 and 30min, respectively. The incidence of clusters in the plasmalemmal vesicles per non-nuclear cytoplasm was 0.01 clusters/m² at 4°C for 2h, increased significantly to 1.08 at 37°C for 5min, and decreased to 0.43 and 0.14 at 15 and 30min, respectively. This work supports that the plasmalemmal invaginations and plasmalemmal vesicles are linked to the HDL uptake in cholesterol-loaded aortic endothelial cells and smooth muscle cells.     

Improvement of β-amylase thermostability in transgenic barley seeds and transgene stability in progeny

The genetic improvement of enzymes important in the brewing process is one of the main goals of barley biotechnology. For the improvement of -amylase thermostability in barley seeds, we have already constructed a mutant thermostable -amylase gene, using site-directed mutagenesis and random mutagenesis to achieve the substitution of seven amino acids of the original barley -amylase. This sevenfold-mutant barley -amylase showed a thermostability increased by 11.6 °C compared to the original enzyme. In the present article, a thermostable -amylase gene under the control of the barley -amylase promoter was introduced to barley protoplasts, and fertile plants were generated from 9 independent transgenic lines. Subsequent analyses indicated that the thermostable -amylase gene was expressed and -amylase activity derived from both native and modified genes was detected in the seeds of 6 transgenic lines. The transgene was stably transmitted to progeny, and thermostable -amylase was synthesized in T₄ seeds, demonstrating that our strategy is applicable for the improvement of seed quality for industrial utilization.     

Purification and immunocytochemical localization of kafirins inSorghum bicolor (L. Moench) endosperm

Summary Kafirins are the storage proteins of sorghum and are found in protein bodies in the seed endosperm. They have been classified as -, -, and -kafirins according to differences in molecular weight, solubility, and structure. The kafirins were purified, amino acid composition was determined, and immunolocalization methods were used to determine the organization of the protein bodies and distribution of kafirins throughout the endosperm. All three groups of kafirins were low in lysine. -Kafirins and -kafirins were relatively high in cysteine, and -kafirins were relatively high in methionine. Transmission electron microscopy showed that protein bodies in the peripheral endosperm were spheroid with concentric rings and few darkly stained inclusions. In contrast, protein bodies of the central endosperm were irregularly shaped with a higher proportion of darkly stained material. The light staining regions of the protein bodies are composed primarily of -kafirins with minor portions of - and -kafirins. The dark staining regions, however, are composed primarily of - and -kafirins. Immunoelectron microscopy showed that protein bodies in the peripheral endosperm contain predominantly a-kafirin with minor amounts of - and -kafirin. Central endosperm protein bodies are also predominantly -kafirin, but have a higher proportion of -kafirin and -kafirin than the peripheral endosperm protein bodies.Abbreviations GAR-HRP Goat anti-rabbit horseradish peroxidase - IgG immunoglobulin G - 2-ME 2-mercaptoethanol - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis - TBS Tris buffer saline - TBS-T Tris buffer saline with Tween - TBS-T-B Tris buffer saline with Tween and bovine serum albumin - TCA trichloroacetic acid - UV ultraviolet     

Micropropagation of candelilla,Euphorbia antisyphilitica Zucc

Axillary shoot proliferation was induced in vitro from shoot explants of greenhouse grown candellila (Euphorbia antisyphilitica Zucc). Optimum shoot proliferation was obtained by supplementing a modified Murashige and Skoog [7] medium with 0.13 M naphthalene-acetic acid and 4.44 M 6-benzylaminopurine. Rooting occurred on 100% of shoots transferred to a medium containing half strength salts supplemented with 0.49 M indole-3-butyric acid. Fully rooted plants were transferred to potting soil and established under greenhouse conditions without special acclimatization techniques.