Transgenic tobacco plants expressing the geminivirus BL1 protein exhibit symptoms of viral disease.

Bipartite geminiviruses, such as squash leaf curl virus (SqLCV), encode two movement proteins (MPs), BR1 and BL1, that are essential for viral movement in and subsequent infection of the host plant. To elucidate the biochemical functions of these MPs and define their respective contributions to viral infection, we have generated transgenic Nicotiana benthamiana plants expressing SqLCV BR1 and BL1. Transgenic plants expressing BR1 or a truncated BL1 were phenotypically indistinguishable from wild-type N. benthamiana. In contrast, transgenic plants expressing full-length BL1, alone or in combination with BR1, were strikingly abnormal both in their growth properties and phenotypic appearance, with leaves that were mosaic and curled under, thus mimicking typical SqLCV disease symptoms in this host. BL1 was localized to the cell wall and plasma membrane fractions, whereas BR1 was predominantly in the microsomal membrane fraction. These findings demonstrate that expression of BL1 in transgenic plants is sufficient to produce viral disease symptoms, and they further suggest that BL1 and BR1 carry out distinct and independent functions in viral movement.     

Management oriented forest map of South India: thematic derivations

The principles and methods of the vegetation mapping undertaken at the French Institute, Pondicherry, are dealt with herein. Particularly, the characterisation of the different types of vegetation and especially the originality of the method: the dynamic interpretation of the vegetation and the depiction of the bioclimatic conditions. The programme of the forest map of South India at scale 1:250 000, undertaken in collaboration with the forest departments of the concerned states, is then described with special attention given to the source and the collection of data. This map has been conceived to serve as a basic document for the sustainable management of the forests. Three examples of its application are given. They concern the detection of anomalies between the existing forest cover and the prevalent climatic environment; the detection of areas for which protection is urgently needed; the selection of regions showing a particular interest in the field of nature conservation or as gene pool reserve. Finally, an example of a thematic map of sensibility of the forests is given, using the vegetation map as a basis.     

Über schlauchförmige Ausstülpungen an der Zellmembran der Makrogametocyten von Eimeria perforans

Summary The surface of the gametocytes and gametes of Eimeria perforans reveals tube-like extrusions which have not been discovered so far in coccidia. These slender tubes are 650 Å in width and at least 1,3 in length. Probably they represent resorbing organelles. The tubes occur only at the surface of older female gametocytes and gametes and have not been observed in merozoites, schizonts, microgametocytes and male gametes. In transversal sections the tubes look like vesicles and are bordered by a membrane the layers of which are not sharply defined. The interior of the tubes seems to be empty after electron microscope observations. In longitudinal sections the membrane of the tubes is striated. The repeating unit of the dark and light bands amounts to about 165 Å. This appearance cannot be explained so far.

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Für beratende Hilfe sind wir Herrn Prof. Dr. R. Danneel und Herrn Prof. Dr. K. E. Wohlfarth-Bottermann zu Dank verpflichtet, für technische Unterstützung Fräulein cand. rer. nat. B. Volkmann. Herrn Dr. D. Spiecker von der Forschungsstelle für Jagdkunde und Wildschadenverhütung, Beuel, danken wir für die Durchführung der Infektionen. Die Mittel für die Untersuchungen stellte uns die Deutsche Forschungsgemeinschaft zur Verfügung.     

Vergleichende Beobachtungen des stomatären Verhaltens verschiedener Pflanzen in Trockenrasen und Laubwäldern am Alpenostrand

Zusammenfassung Innerhalb der drei unterschiedenen Vegetationseinheiten: Trockenrasen+Felssteppen, Trockenwälder und mesophiler Laubwald wurden die Arten mit Hilfe der Infiltrationsflüssigkeiten Paraffinöl, Alkohol und Xylol auf ihr stomatäres Verhalten während der ganzen Vegetationsperiode untersucht und in die drei Verhaltenstypen starke, schwache und wechselnde Schließtendenz eingeordnet. Um standörtliche Unterschiede herauszuarbeiten, wurde aus den an einem bestimmten Wuchsort während einer bestimmten Zeit an möglichst vielen verschiedenen Pflanzen gewonnenen Infiltrationswerten jeweils ein Infiltrations-Mittelwert gebildet und außerdem, sofern Tages- und Nachtwerte von demselben Wuchsort vorlagen, auch ein Quotient aus (womöglich) höchstem Infiltrations-Mittelwert des Tages durch (womöglich) tiefsten Infiltrations-Mittelwert der Nacht (Q _T Q _N ) gebildet, um damit die Intensität der Spaltenbewegung an verschiedenen Standorten zu erfassen. Es ergab sich im allgemeinen bei trockenem Wetter und an trockenen Standorten ein intensiveres Spaltenspiel mit stärkerer nächtlicher Schließtendenz als bei feuchterem Wetter bzw. an feuchteren Standorten. Ausnahmen werden auf S. 598 erörtert.Was die standörtliche Verteilung der stomatären Verhaltenstypen betrifft, so überwiegen in der untersuchten Vegetation des behandelten Gebietes überall die Arten mit wechselnder Schließtendenz, während solche mit schwacher nächtlicher Schließtendenz im Trockenrasen überhaupt fehlen und auch im Wald nur eine sehr geringe Rolle spielen, im Gegensatz zu feuchteren Gebieten bzw. Standorten. Die Arten mit starker nächtlicher Schließtendenz sind im Trockenrasen verhältnismäßig am häufigsten.     

Physiologische Untersuchungen an einer ungewöhnlich säureresistenten Chlorella

Summary The influence of the hydrogen-ion concentration on the growth and metabolism of a highly acid-resistant green alga, Chlorella ellipsoidea (strain Marburg St), was studied. Chlorella pyrenoidosa (Emerson strain) served as a normal control organism. Growth of Chlorella ellipsoidea occurs in the entire range from Ph 2.0 to Ph 10, whereas for Chlorella pyrenoidosa the limits were found to be Ph 3.5 and Ph 10. Respiration is much less sensitive to hydrogen-ion concentration in the acid-resistant as compared to the normal strain. Thus an increase in acidity from Ph 4.0 to Ph 2.0 increases the respiratory oxygen uptake by 120% in Chlorella pyrenoidosa and by 25% in Chlorella ellipsoidea. In addition, only the less resistant Chlorella pyrenoidosa shows an accumulation of nitrite in the dark in acid culture media, indicating a disturbance of the normal course of nitrate reduction under these conditions. On the other hand, the rate of photosynthesis of both organisms was found to be almost independent of acidity between Ph 4.0 and Ph 2.0. At the acid and alkaline limits of growth in both algae, an inhibition of cell division leads to an increase of cell size and dry weight per cell, frequently connected with the occurrence of bizarre giant cells. — In addition, adaptation phenomena were found to play a role in determining the acid limit of growth. Cells of Chlorella ellipsoidea, after inoculation from normal medium (Ph about 6) into a solution of Ph 2.0, begin growth at a high rate only after a lag of about two weeks. Cells grown previously in an acid medium, however, immediately resume growth upon inoculation into a medium of Ph 2.0. This adaptation involves a considerable reduction of cell size.     

Endo's stress analysis of the primate skull and the functional significance of the supraorbital region

A review of Endo's experimental and theoretical procedures and data indicates that the magnitude of the principal strains in the glabella region of both humans and gorillas are low as compared to other parts of the face. Therefore, his data do not provide support for the hypothesis that the glabella region is a highly stressed region during biting. In addition, increased levels of strain in the supraorbital region are directly related to increased levels of masticatory muscle and reaction forces, and not necessarily to anterior tooth loading as opposed to posterior tooth loading. His data also indicate that the supraorbital region in extant humans cannot be accurately modeled as a beam. These conclusions either differ from those of Endo or are not clearly presented or emphasized throughout any of Endo's papers. Therefore, we suggest that a number of investigators have made unsupported or erroneous conclusions based on Endo's work. This is particularly true for those studies that have emphasized the existence of powerful bending stress in the glabella region during incisor biting in both humans and non-human primates.     

First chemical synthesis of a scorpion alpha-toxin affecting sodium channels: the Aah I toxin of Androctonus australis hector.

Aah I is a 63-residue alpha-toxin isolated from the venom of the Buthidae scorpion Androctonus australis hector, which is considered to be the most dangerous species. We report here the first chemical synthesis of Aah I by the solid-phase method, using a Fmoc strategy. The synthetic toxin I (sAah I) was renatured in DMSO-Tris buffer, purified and subjected to thorough analysis and comparison with the natural toxin. The sAah I showed physico-chemical (CD spectrum, molecular mass, HPLC elution), biochemical (amino-acid composition, sequence), immunochemical and pharmacological properties similar to those of the natural toxin. The synthetic toxin was recognized by a conformation-dependent monoclonal anti-Aah I antibody, with an IC50 value close to that for the natural toxin. Following intracerebroventricular injection, the synthetic and the natural toxins were similarly lethal to mice. In voltage-clamp experiments, Na(v) 1.2 sodium channel inactivation was inhibited by the application of sAah I or of the natural toxin in a similar way. This work describes a simple protocol for the chemical synthesis of a scorpion alpha-toxin, making it possible to produce structural analogues in time.     

Cloning and characterization of theeapB andeapC genes ofCryphonectria parasitica encoding two new acid proteinases,and disruption ofeapC

Two new proteinases secreted byCryphonectria parasitica, namely EapB and EapC, have been purified. The corresponding structural genes were isolated by screening a cosmid library, and sequenced. Comparison of genomic and cDNA sequences revealed that theeapB andeapC genes contain three and two introns, respectively. The products of theeapB andeapC genes as deduced from the nucleotide sequences, are 268 and 269 residues long, respectively. N-terminal amino acid sequencing data indicates that EapC is synthesized as a zymogen, which yields a mature 206-amino acid enzyme after cleavage of the prepro sequence. Similarly, sequence alignment studies suggest that EapB is secreted as a 203-residue form which shares extensive similarities not only with EapC but also with two other acid fungal proteinases. However, they display distinct structural features; for example, no cysteine residue is found in EapC. TheeapC gene was mutated using a two-step gene replacement strategy which allowed the specific introduction of several stop codons at the beginning of theeapC coding sequence in an endothiapepsin-deficient (EapA⁺)C. parasitica strain. Although the resulting strain did not secrete EapC, it still exhibited residual extracellular proteolytic activity, which could be due to EapB.     

Subsets of sialylated, sulfated mucins of diverse origins are recognized by L-selectin. Lack of evidence for unique oligosaccharide sequences mediating binding

Previous studies have shown that the mucin-type polypeptidesGlyCAM-1, CD34, and MAdCAM-1 can function as ligands for L-selectinonly when they are synthesized by the specialized high-endothelialvenules (HEV) of lymph nodes. Since sialylation, sulfation,and possibly fucosylation are required for generating recognition,we reasoned that other mucins known to have such componentsmight also bind L-selectin. We show here that soluble mucinssecreted by human colon carcinoma cells, as well as those derivedfrom human bronchial mucus can bind to human L-selectin in acalcium-dependent manner. As with GlyCAM-1 synthesized by lymphnode HEY, 2–3 linked sialic acids and sulfation seem toplay a critical role in generating this L-selectin binding.In each case, only a subset of the mucin molecules is recognizedby L-selectin. Binding is not destroyed by boiling, suggestingthat recognition may be based primarily upon carbohydrate structures.Despite this, O-linked oligosaccharide chains released fromthese ligands by beta-elimination do not show any detectablebinding to L-selectin. Following protease treatment of the ligands,binding persists in a subset of the resulting fragments, indicatingthat specific recognition is determined by certain regions ofthe original mucins. How ever, O-linked oligosaccharides releasedfrom the subset of non-binding mucin fragments do not show verydifferent size and charge profiles compared to those that dobind. Furthermore, studies with polylactosamine-degrading endoglycosidasessuggest that the core structures involved in generating bindingcan vary among the different ligands. Taken together, thesedata indicate that a single unique oligosaccharide structuremay not be responsible for high-affinity binding. Rather, diversemucins with sialylated, sulfated, fucosylated lactosamine-typeO-linked oligosaccharides can generate high-affinity L-selectinligands, but only when they present these chains in unique spacingand/or clustered combinations, presumably dictated by the polypeptidebackbone. L-selectin mucins sialic sialic acid sulfate adhesion