Separation of plant membranes by electromigration techniques

The review focuses on the multiple separating regimes that offers the free flow electrophoresis technique: free flow zone electrophoresis, isoelectric focusing, isotachophoresis, free flow step electrophoresis. Also, the feasibility to apply either interval or continuous flow electrophoresis is evaluated. The free flow zone electrophoresis regime is generally selected for the separation of cells, organelles and membranes while the other regimes find their largest fields of applications in the purification of proteins and peptides. The latter regimes present the highest resolution efficiency. Therefore, a large part of this review is devoted to the applicabilities of these different regimes to the purification of organelles and membrane vesicles at the preparative scale. Recent developments, both in instrumentation and procedures, are described. The major achievements in plant membrane fractionation obtained with free flow electrophoresis are outlined. The related procedures are both analytical and preparative: they separate tonoplast and plasma membrane simultaneously from the same homogenate, they discriminate for one type of membrane vesicles of opposite orientation, and process large quantities of membrane material by reason of the continuous flow mode. Recent advances using electromigration techniques that permit confirmation of the dynamic state of membranes, characterisation of complex membrane-dependent functions and discovery of new membrane-localised activities are presented.     

Studies on immunoassays of peptide factors. IV. New synthesis of a gastrin/peroxidase conjugate

We have shown that structurally well-defined homogeneous maleoyl-peptides are synthetically accessible. These anchor-modified peptide derivatives allow their selective covalent linkage to thiol-containing proteins via the maleimide-thiol procedure. Correspondingly mercaptosuccinylated horseradish peroxidase was reacted with N alpha-maleoyl-beta-alanyl-human-little gastrin-I-[2-17] to produce the gastrin/peroxidase conjugate in good yields at 1:1 stoichiometry. The conjugate exhibited full enzymatic activity and identical binding affinity to antigastrin antisera as the parent gastrin. This approach proved to be well suited for the preparation of enzyme labeled peptide factors as tracers for immunoassays.     

Cloning and nucleotide sequence of the chlD locus

The nucleotide sequence of a Sau3A1 restriction nuclease fragment that complemented an Escherichia coli chlD::Mu cts mutant strain was determined. DNA and deduced amino acid sequence analysis revealed two open reading frames (ORFs) that potentially codes for proteins with amino acid sequence homology with binding protein-dependent transport systems. One of the ORFs showed a sequence that encoded a protein with properties that were characteristic of a hydrophobic inner membrane protein. The other ORF, which was responsible for complementing a chlD mutant, encoded a protein with conserved sequences in nucleotide-binding proteins and hydrophilic inner membrane proteins in active transport systems. A proposal that the chlD locus is the molybdate transport operon is discussed in terms of the chlD phenotype.     

Substrate specifity of the hexose carrier in the plasmalemma of Chenopodium suspension cells probed by transmembrane exchange diffusion

Substrate specifity of the proton-driven hexose cotransport carrier in the plasmalemma of photoautotrophic suspension cells of Chenopodium rubrum L. has been studies through the short-term perturbation of ¹⁴C-labelled efflux of 3-O-methyl-d-glucose. Efflux, occurring exclusively via carrier-mediated exchange diffusion, is trans-stimulated by the substrate and trans-inhibited by the glucose-transport inhibitors phlorizin (K _1/2=7.9 mM) and its aglucon phloretin (K _1/2=84 μM); with both inhibitors, 3-O-methyl-d-glucose efflux may be blocked completely. Trans-stimulation of efflux (up to fourfold) by a variety of the d-enantiomers of neutral hexoses, including glucose (K _1/2=48 μM), 3-O-methyl-d-glucose (K _1/2=139 μM), and fructose (K _1/2=730 μM), but not by, for instance, d-allose, and l-sorbose, shows that carrier-substrate interaction critically involves the axial position at C-1 and C-3, respectively. We suggest that substrate binding by the Chenopodium hexose carrier involves both hydrophobic interaction with the pyran-ring and hydrogen-ion bonding at C-1 and C-3 of the d-glucose conformation.     

Development of a liquid flow-through system to inhibit browning in callus cultures of guayule (Parthenium argentatum Gray)

Calli of P. argentatum were grown on a newly designed liquid nutrient flow-through system which facilitated the subculturing of calli and delayed browning for 6 weeks. Friable calli were obtained on half-strength Gamborg B5-medium supplemented with 0.05 mgl⁻¹ 2,4-dichlorophenoxyacetic acid. Shoots developed on media supplemented with 0.2 mgl⁻¹ benzylaminopurine but lacking 2,4-dichlorophenocyacetic acid.     

Structure fine du rein de l'Epinoche (Gasterosteus aculeatus L.) au cours de sa transformation muqueuse

Résumé Pendant la période de reproduction, les néphrons du rein de l'Epinoche mâle subissent d'importantes modifications de structure sous l'action des hormones sexuelles. Au niveau de chacun d'entre eux, se différencient deux segments distincts par leur fonction et par leur cytologie. Le segment urinaire, très court, est formé de cellules identiques à celles du jeune, qui remplissent leur fonction d'excrétion. Le segment glandulaire, plus volumineux, subit une transformation muqueuse et élabore une sécrétion qui sert à construire le nid. L'évolution de ces deux segments est étudiée au cours de la période de reproduction et les modifications cytologiques correspondantes sont décrites.

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Fine structure of the kidney of the three-spined-stickleback (Gasterosteus aculeate L.) during its mucous transformation

Summary Under the action of sexual hormones the nephrons of the kidney of the male three-spined-stickleback undergo considerable transformations during the breeding period. They differentiate into two segments which differ from one another in function and cytology. The cells of the urinary segment are identical to those of the young fish. They have an excretory function. The glandular segment undergoes a mucous transformation and synthesizes a secretion which is used for the building of the nest. The cytological transformations occuring at the level of these two segments during the breeding period are described with special attention to the mucous cells.

     

Über das gleichzeitige Vorkommen von zweierlei Eiweißkörpern in den Zellkernen vonPseudolysimachion spicatum und einigen anderen Scrophulariaceen

Zusammenfassung Die Zellkerne vonPseudolysimachion spicatum, Veronica cymbalaria undV. gentianoides enthalten zweierlei Eiweißkörper von je nach dem Entwicklungszustand unterschiedlicher Konsistenz. Bis zum Höhepunkt ihrer Entwicklung liegen sie als anscheinend amorphe flüssige oder gelartige Gebilde vor. Später tritt — mit gewebespezifischen Unterschieden — Verfestigung und Kristallisation ein, und zwar bei beiden nicht gleichzeitig und z. T. auch nur beim großen. Beiderlei Körper unterscheiden sich deutlich in physikalischer und chemischer Hinsicht, außerdem in ihrer Entwicklungsgeschichte und Strukturveränderung. Sie fehlen in den Schließzellen, im Antherentapetum, in den Pollenmutterzellen, Pollenkörnern, im Embryosack, im Endosperm und im Embryo.Die Zellkerne in der Blattepidermis vonPenstemon barbatus enthalten ebenfalls zweierlei Eiweißkörper. Davon liegt einer als kugeliges Gebilde, der andere als Kristallstapel vor.

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Summary In the nuclei ofPseudolysimachion spicatum, Veronica cymbalaria, V. gentianoides there occur two different kinds of protein bodies. They clearly differ from one another in physical and chemical regard as well as in their ontogeny. At first they are amorphous, liquid or geluous. As a rule a consolidation or crystallization with differences according to the tissue takes place after the climax of the development, and that not at the same time within both bodies. In the stomata cells, pollen mother cells, in the embryo sac, in the endosperm, in the endothelium and in the embryo both bodies do not occur.The nuclei in the epidermis of the leaves ofPenstemon barbatus also contain two different kinds of protein bodies, one of them appearing as a round body, and the other one as a pile of crystal plates.

     

Temperature influence on cohort parameters and demographic characteristics of the two cowpea coreids Clavigralla tomentosicollis and C. shadabi

Age-specific life tables of two important pests of cowpea, Vigna unguiculata (L.) Walp., the pod sucking bugs Clavigralla tomentosicollis Stål and C. shadabi Dolling (Heteroptera: Coreidae), were obtained from observations carried out at different temperatures. A biophysical model was found satisfactory to describe the temperature-response of developmental and mortality rates of egg and nymphal stages, with a peak developmental rate around 34°C in both species. The variability in development times was small and the experimental data did not permit any conclusion with regard to the Erlang probability density function. Survival of eggs and nymphs remained high between 20° and 30°C for both species. At temperatures above 34°C, C. tomentosicollis survivorship and fecundity was higher than that of C. shadabi, which in turn laid more eggs at temperatures between 20° and 30°C. Maximum fecundity is estimated to be at 29°C for C. tomentosicollis (99 eggs/female) and 26°C for C. shadabi (261 eggs/female). At 30°C, the intrinsic rate of increase reached a maximum in both species, 0.152 per day for C. tomentosicollis and 0.145 per day for C. shadabi, and remained high for C. tomentosicollis until 36°C. C. tomentosicollis performed significantly better on pigeonpea, Cajanus cajan Millsp., than on cowpea at higher temperatures.