Phylogeny of Greya (Lepidoptera: Prodoxidae), based on nucleotide sequence variation in mitochondrial cytochrome oxidase I and II: congruence with morphological data

The phylogeny of Greya Busck (Lepidoptera: Prodoxidae) was inferred from nucleotide sequence variation across a 765-bp region in the cytochrome oxidase I and II genes of the mitochondrial genome. Most parsimonious relationships of 25 haplotypes from 16 Greya species and two outgroup genera (Tetragma and Prodoxus) showed substantial congruence with the species relationships indicated by morphological variation. Differences between mitochondrial and morphological trees were found primarily in the positions of two species, G. variabilis and G. pectinifera, and in the branching order of the three major species groups in the genus. Conflicts between the data sets were examined by comparing levels of homoplasy in characters supporting alternative hypotheses. The phylogeny of Greya species suggests that host-plant association at the family level and larval feeding mode are conservative characters. Transition/transversion ratios estimated by reconstruction of nucleotide substitutions on the phylogeny had a range of 2.0-9.3, when different subsets of the phylogeny were used. The decline of this ratio with the increase in maximum sequence divergence among taxa indicates that transitions are masked by transversions along deeper internodes or long branches of the phylogeny. Among transitions, substitutions of A-->G and T-->C outnumbered their reciprocal substitutions by 2-6 times, presumably because of the approximately 4:1 (77%) A+T-bias in nucleotide base composition. Of all transversions, 73%-80% were A<-->T substitutions, 85% of which occurred at third positions of codons; these estimates did not decrease with an increase in maximum sequence divergence of taxa included in the analysis. The high frequency of A<-->T substitutions is either a reflection or an explanation of the 92% A+T bias at third codon positions.      

A new method for the determination of the anionic adsorption capacity of bottom deposits with particular reference to the adsorption of anionic detergents

Summary After a short discussion of the scarce and incomplete literature on anionic adsorption properties of soils and sediments, a new technique based on the adsorption of manoxol to soils, sediments and pure humic acid is described.Adsorption curves for various sediments, humic acid and clays are given. The adsorption capacity of bottom deposits are interpreted in terms of humic acid equivalents.The method can be used for series analysis of both the specific adsorption capacity for manoxol of river muds and also as a more general parameter for the anionic and surface adsorption capacity of various materials such as sludges, soils and compost.

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Zusammenfassung Nach einer kurzenBesprechung der bestehenden spärlichen Literatur über anionische Adsorptionseigenschaften von Böden und Sedimenten wird eine Technik, die auf der Adsorption van Manoxol (Na-dioctyl-sulfosuccinat) an Bodensedimenten beruht, beschrieben.Adsorptionskurven verschiedener Sedimente, Huminsäuren und einiger Tone werden graphisch dargestellt und die Adsorptionskapazitäten von Bodensedimenten als Huminsäure-äquivalente ausgedrückt und interprätiert.Die beschriebene Methode eignet sich für Serienanalysen zur Bestimmung der spezifischen Adsorptionskapazität für Manoxol von Sedimenten natürlicher Gewässer und kann auch als ein allgemeiner Parameter zur Charakterisiering der anionischen und Oberflächen-adsorptionskapazität von Substraten, wie Schlamm, Böden und Kompost angewandt werden.

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National Institute for Water Research, S. A. Council for Scientific and Industrial Research     

Characterization of endosome-endosome fusion in a cell-free system using Dictyostelium discoideum.

An in vitro endosome fusion assay using Dictyostelium discoideum is described. The method requires endocytosis of anti-dinitrophenol (DNP) IgG or DNP-derivitized beta-glucuronidase into two sets of cells. After homogenizing the cells, the vesicles were mixed, and fusion was measured by quantitating immune complex formation between DNP-beta-glucuronidase and anti-DNP IgG. Fusion was dependent upon ATP, temperature, pH, ionic strength, and cytosol and sensitive to detergent, dilution, trypsin, N-ethylmaleimide, and guanosine 5'-3-O-(thio)triphosphate. Although weak bases, ionophores, hadacidin, [ethylenebis(oxyethylenenitrilo)]tetraacetic acid, and caffeine inhibit endocytosis in vivo, these reagents had no affect on in vitro endosome fusion. Comparison of Dictyostelium with mammalian cells showed differences in the temperature, pH, and salt requirements for fusion, possibly reflecting differences in the life-styles of various cell types. Like mammalian cells, Dictyostelium required GTP-binding protein(s) and an N-ethylmaleimide-sensitive factor for endosome fusion. Thus, the mechanism driving endosome fusion may have been conserved throughout evolution. Electron microscopic studies confirmed in vitro endosome fusion and revealed endosomes were being engulfed by other endosomes, resulting in formation of multivesicular elements (i.e. autophagic vesicles). This system may be useful for characterizing mutations, evolution, and developmental regulation along the endocytic pathway.