Functional expression of the Erwinia uredovora carotenoid biosynthesis gene crtl in transgenic plants showing an increase of β-carotene biosynthesis activity and resistance to the bleaching herbicide norflurazon

Among the enzymes involved in carotenoid biosynthesis, phytoene desaturase is considered to be a rate-limiting enzyme in this pathway and is also the target of many bleaching herbicides. This enzyme shows diversity concerning its function and amino acid homology among various organisms. The phytoene desaturase gene crtl of Erwinia uredovora was expressed, the 5'-region of which was fused to the sequence for the transit peptide of a pea Rubisco small subunit, in tobacco plants under the control of the CaMV 35S promoter. This chimeric gene product was targeted into chloroplasts and processed in the transgenic plants. The production and processing of the corresponding protein could be demonstrated by Western blotting. Immunogold localization showed that the location of the gene product Crtl was preferentially in the thylakoids. A radioactive labeling study using the leaves demonstrated enhanced activity for the synthesis of β-carotene. In addition, the transgenic tobacco acquired elevated resistance to the bleaching herbicide norflurazon.     

Correlation between arrested secondary plasmodesmal development and onset of accelerated leaf senescence in yeast acid invertase transgenic tobacco plants

Mature leaves of a transgenic tobacco plant ( Nicotiana tabacum L var. Samsun, line A41-10) that constitutively express a yeast-derived acid invertase gene develop symptoms which are characterized by the presence of greenish-yellow and green sectors in the same leaf, and onset of early, leaf senescence. Previous studies indicated that invertase activity was two- to threefold higher in the greenish-yellow sectors than in the green sectors. Our structural analyses revealed that development of secondary plasmodesmata, via modification of existing primary plasmodesmata, between mesophyll cells was inhibited severely in the greenish-yellow sectors, but only marginally in the green sectors. In contrast, the structure and function of primary plasmodesmata in the same symptomatic sectors remained unaltered as determined by structural and dye coupling studies. It is hypothesized that secondary plasmodesmata differ from primary plasmodesmata in having special abilities to traffic information molecules to coordinate leaf development and physiological function(s). Arrest of secondary plasmodesmal development by high invertase activity in the transgenic tobacco leaf may have prevented this type of trafficking and hence resulted in early leaf senescence. The results also indicate that the yeast acid invertase-expressing tobacco may provide an effective experimental system for the molecular characterization of cellular mechanisms that regulate the development, function, and possible turnover of secondary plasmodesmata.     

Natural lignin modulators improve bagasse saccharification of sugarcane and energy cane in field trials

The burgeoning cellulosic ethanol industry necessitates advancements in enzymatic saccharification, effective pretreatments for lignin removal, and the cultivation of crops more amenable to saccharification. Studies have demonstrated that natural inhibitors of lignin biosynthesis can enhance the saccharification of lignocellulose, even in tissues generated several months post-treatment. In this study, we applied daidzin (a competitive inhibitor of coniferaldehyde dehydrogenase), piperonylic acid (a quasi-irreversible inhibitor of cinnamate 4-hydroxylase), and methylenedioxy cinnamic acid (a competitive inhibitor of 4-coenzyme A ligase) to 60-day-old crops of two conventional Brazilian sugarcane cultivars and two energy cane clones, bred specifically for enhanced biomass production. The resultant biomasses were evaluated for lignin content and enzymatic saccharification efficiency without additional lignin-removal pretreatments. The treatments amplified the production of fermentable sugars in both the sugarcane cultivars and energy cane clones. The most successful results softened the most recalcitrant lignocellulose to the level of the least recalcitrant of the biomasses tested. Interestingly, the softest material became even more susceptible to saccharification.     

Über die Einwirkung Hypotonischer Medien auf das Zwischenhirn-Hypophysensystem von Mugil und Gobius

Zusammenfassung Bei den Teleostiern Mugil und Gobius wurden die Veränderungen des Zwischenhirn-Hypophysensystems in veränderten Außenmedien (hypotonisches Seewasser) histologisch untersucht.Beim langsamen Überführen aus Seewasser von 38,4 S in Seewasser von 25,4 und 20,8 S nahm das Sekret nicht, wie erwartet, zu. Im Nucleus praeopticus und im Hinterlappen der Hypophyse erfolgt eine starke Verringerung des Sekretbestandes (Mugil). Erst in Seewasser von weit niedrigerem Salzgehalt (6,3 und 1, 4) nimmt das Sekret im Hypophysenhinterlappen wieder zu (Gobius). Bei 6 S ist eine intensive Sekretableitung im N.p. und Tractus praeopticohypophyseus zu beobachten. Eine Deutung der Ergebnisse wird versucht, indem das weitgehend konstante Innenmedium mit den wechselnden Außenfaktoren in Beziehung gebracht wird.Die Befunde weisen darauf hin, daß das Zwischenhirn-Hypophysensystem auch bei Teleostiern an der Regulation des Wasserhaushaltes beteiligt ist.Der Deutschen Forschungsgemeinschaft danke ich für die Gewährung eines Stipendiums, das mir den Aufenthalt in Neapel ermöglichte. Mein besonderer Dank gilt auch den Herren Prof. Dr. Remane, Prof. Dr. Bargmann und Dozent Dr. Schiebler für ihre Unterstützung.     

Untersuchungenüber Das tageszeitlich gebundene Schlüpfen yon Pseudosmittia Arenaria (Dipt. Chironomidae)

Zusammenfassung In einleitenden terminologischen Erwägungen wird vorgeschlagen, den Ausdruck Schlüpfrhythmus zugunsten von tageszeitlich gebundenem Schlüpfen aufzugeben. Pseudosmittia arenaria zeigt im normalen Tag-Nachtwechsel ein deutliches tageszeitlich gebundenes Schlüpfen mit einem Maximum 6–8h nach DL (Beginn der Beleuchtung).In Dauerdunkel schlüpfen keine Imagines. In Dauerlicht erscheinen die Tiere gleichmäßig über den ganzen Tag verteilt.Der ganze Bereich des sichtbaren Lichtes (geprüft von 476–641 m) ist wirksam. Auch die Lichtintensität in Licht-Dunkelbedingungen spielt im untersuchten Bereich (18–350 Lux) keine Rolle.Das Schlüpfmaximum von Ps. arenaria zeigt eine deutliche Beziehung zu DL. Es wandert bei konstanter Tageslänge um so dichter an DL heran, je kürzer die relative Länge der Lichtzeit ist.Bei kürzeren Tageslängen wandert das Maximum von DL fort, bei längeren an DL heran. Bei Tagen, die kürzer als 18h sind, liegt es in der folgenden Dunkelzeit, bei solchen, die länger als 36h sind, vor DL. Sein Weg beschreibt dabei eine kubische Parabel.Beim Eintritt des Gipfels in die folgende Dunkelzeit erscheint ein Maximum nur an jedem 2. Tag. Ein Maximum vor DL ruft ein zusätzliches Maximum in der gleichen Entfernung von LD (Beginn der Dunkelzeit) hervor. Bei extremen Tageslängen ist also dennoch ein Maximum ungefähr alle 24h zu beobachten. Dies ist exogen bedingt. Verlagert man durch schwache Beleuchtung während der Dunkelzeit das Maximum im 12h-Tag in die Lichtzeit, so erhält man ein Maximum an jedem Tag. Der Eintritt des Maximums in eine Dunkelzeit hat also das Umspringen auf einen Schlüpfgipfel an jedem 2. Tag bzw. auf 2 je Tag zur Folge.Im 12h-Tag mit absoluter Dunkelzeit während der Dunkelzeit kann man auch ein Maximum alle 12h erreichen, und zwar durch Umstimmung eines Teils der Population. Man hat dann 2 Gruppen, deren Maxima um 12h gegeneinander verschoben sind. Hemmend und fördernd auf den Einfluß des Lichtes wirken Temperatur und Substratfeuchtigkeit. Beide Faktoren können ohne Licht kein Schlüpfen hervorrufen. Beziehungen zwischen Lebensweise, Ökologie oder systematischer Stellung und tageszeitlich gebundenem Schlüpfen lassen sich bisher nicht feststellen.     

Utilization of metabolic scope in relation to feeding and activity by individual and grouped zebrafish, Brachydanio rerio (Hamilton-Buchanan)

Metabolic scope and its utilization in relation to feeding and activity were measured in individual and grouped zebrafish (weight range, 430–551 mg) at 24° C by respirometry. Mean maximum metabolic rate, induced by swimming to exhaustion, R_max(i), was 1223 (s.d. , 157) mg O₂, kg^?1 h^?1 for individuals. Standard metabolic rate, R_s. was 364 mg O₂ kg^?1 h^?1, as estimated by extrapolating to zero activity from measurements of unfed, spontaneously active individuals. Mean routine metabolic rate, R_rout, of individuals was 421 (s.d. , 58) mg O₂, kg_-1 h_-1. The mean voluntary maximum metabolic rate, R_max(v), following transfer of minimally exercised fish to the respirometer, was 1110 (s.d. , 83) mg O₂ kg ^?1 h^?1 for groups of six fish, and was not significantly different from the value measured for individuals, 1066 (s.d. , 122) mg O₂, kg^?1 h^?1. Grouped fish acclimated to the respirometer more slowly than individual fish and exhibited significantly higher R_rout, apparently a result of greater social interaction and activity in groups. Mean R_rout for groups was 560 (s.d. , 78) mg O₂, kg^?1 h^?1. While groups of zebrafish fed a ration of 5% wet body weight day^?1 exhibited consistently higher metabolic rates than fish fed rations of 2.5% wet body weight day^?1 the high ration group still used only a maximum of 77% of the metabolic scope. Zebrafish of the size studied do not appear to demonstrate a high degree of conflict in utilization of metabolic scope by different respiratory components. The metabolic rates measured for zebrafish are among the highest yet measured for fish of similar size and at similar temperatures.     

THE FINE STRUCTURE OF TWO PHOTOSYNTHETIC SPECIES OF DINOPHYSIS (DINOPHYSIALES,DINOPHYCEAE)1

Two closely related, photosynthetic species belonging to the genus Dinophysis were examined, D. acuminata Claparède et Lachmann and D. fortii Pavillard. Typical dinoflagellate features include the amphiesmal covering enclosing the cells and the structure of the nucleus and mitochondria. Many other characteristics seem to be specific to the order Dinophysiales. Many rhabdosomes are present, and complex mucocysts are found beneath the amphiesma. The thecal pores are unusual with the base of the pore occluded by a thin disc that is continuous with the main amphiesmal plate. The structure of the apical pore is also distinctive. Chloroplasts are grouped together in chromatospheres, enclosed by a double membrane, and contain paired thylakoids with electron dense contents in the lumen. The two pusules are extensive, each branching off the flagellar canal, and consisting of a large antechamber and a number of convoluted sacs. The entrance of each antechamber, and site of an emerging flagellum, is surrounded by a striated fibrous collar. Near the flagellar pore is a prominent microtubular/microbody complex which penetrates deep into the cell cytoplasm. Consideration is given to taxonomic position of the Dinophysiales and also to the nature and origins of the chloroplasts.     

First report on new evidence for the occurrence of Podocarpus and possible human presence at the mouth of the Amazon during the Late-glacial

Palynological studies on late Quaternary lake sediments from the region of the Amazon estuary, 100 km north-east of Belém, Pará State, Brazil, enable reconstruction of lowland Amazonian rain forest during the Late-glacial and Holocene periods. Late-glacial forests included populations of Podocarpus which suggests a distinct climatic cooling. Ilex was abundant in the early Holocene. Records of the mangrove taxon, Rhizophora, indicate rapid Atlantic sea-level rise in the beginning of the Holocene. High charcoal representation may reflect the first arrival of Amerindians in the Amazon coastal area, probably about 10 800 B.P.     

Absence of plasma membrane H+-ATPase in plasmodesmata located in pit-fields of the young reactive pulvinus ofMimosa pudica L.

Summary Immunocytochemical techniques were employed to study the spatial distribution of the plasma membrane H⁺-ATPase within various cell types of the young reactive primary pulvinus ofMimosa pudica L. These cells were interconnected by large numbers of plasmodesmata, being concentrated within pit-fields. Although we could routinely detect evidence of the H⁺-ATPase along the plasma membrane, immunolabelling was rarely, if ever, observed along the plasma membranes of the plasmodesmata. This finding is discussed with respect to the likely specialized supramolecular structure of the plasmodesma.Abbreviations SEL size exclusion limit of plasmodesmata     

snRNP Sm proteins share two evolutionarily conserved sequence motifs which are involved in Sm protein-protein interactions.

The spliceosomal small nuclear ribonucleoproteins (snRNPs) U1, U2, U4/U6 and U5 share eight proteins B', B, D1, D2, D3, E, F and G which form the structural core of the snRNPs. This class of common proteins plays an essential role in the biogenesis of the snRNPs. In addition, these proteins represent the major targets for the so-called anti-Sm auto-antibodies which are diagnostic for systemic lupus erythematosus (SLE). We have characterized the proteins F and G from HeLa cells by cDNA cloning, and, thus, all human Sm protein sequences are now available for comparison. Similar to the D, B/B' and E proteins, the F and G proteins do not possess any of the known RNA binding motifs, suggesting that other types of RNA-protein interactions occur in the snRNP core. Strikingly, the eight human Sm proteins possess mutual homology in two regions, 32 and 14 amino acids long, that we term Sm motifs 1 and 2. The Sm motifs are evolutionarily highly conserved in all of the putative homologues of the human Sm proteins identified in the data base. These results suggest that the Sm proteins may have arisen from a single common ancestor. Several hypothetical proteins, mainly of plant origin, that clearly contain the conserved Sm motifs but exhibit only comparatively low overall homology to one of the human Sm proteins, were identified in the data base. This suggests that the Sm motifs may also be shared by non-spliceosomal proteins. Further, we provide experimental evidence that the Sm motifs are involved, at least in part, in Sm protein-protein interactions. Specifically, we show by co-immunoprecipitation analyses of in vitro translated B' and D3 that the Sm motifs are essential for complex formation between B' and D3. Our finding that the Sm proteins share conserved sequence motifs may help to explain the frequent occurrence in patient sera of anti-Sm antibodies that cross-react with multiple Sm proteins and may ultimately further our understanding of how the snRNPs act as auto-antigens and immunogens in SLE.