Taxonomic studies of the genus Hypoxis in East Africa

East African material of the genus Hypoxis L. has preliminarily been divided into the heterogenous, probably apomictic H. obtusa Burch- complex (2n = 40–50, ca. 75, 76, ca. 85, >86, ca. 92, ca. 98, ca. 108, 130–135, 160–200) and 5 rather homogenous species: H. angustifolia Lam. (2n = 14, 28), H. goetzei Harms (2n = ca. 62), H. kilimanjarica Bak., H. malosana Bak. (2n = 14) and H. macrocarpa Holt & Staubo sp. nov. H. kilimanjarica is divided into ssp. kilimanjarica and ssp. prostrata Holt & Staubo ssp. nov.     

Involvement of the cytoplasmic membrane in nitrogen fixation by Rhizobium leguminosarum bacteroids

1. The nitrogen-fixing efficiency of freshly prepared suspensions of Rhizobium leguminosarum bacteroids from pea root nodules was considerably enhanced by addition of bovine serum albumin. Evidence was found that during preparation of bacteroids the cell membrane is exposed to the uncoupling effect of free fatty acids and to plant phospholipase D activity. Both effects could be counteracted by bovine serum albumin. 2. A technique was developed by which concentrations of free O2 and nitrogenase activity could be measured simultaneously under conditions of steady-state respiration. By means of this system it could be shown that in contrast to previous claims, high ATP/ADP ratios can be achieved in bacteroids even with a high concentration of O2 in the medium. 3. Nitrogen fixation was found to be controlled by the ATP/ADP ratio, the generation of reducing equivalents and the switch-off phenomenon. It was demonstrated that the generation of reducing equivalents for nitrogenase is regulated by the energized state and the integrity of the bacteroid cell membrane. The data indicate that the process of aerobic nitrogen fixation in R. leguminosarum bacteroids resembles that of Azotobacter vinelandii.     

Antioxidant activities of carotenoids: quantitative relationships between theoretical calculations and experimental literature data

Quantitative structure activity relationships (QSARs) are described for the antioxidant activity of series of all-trans carotenoids. The antioxidant activity of the carotenoids is characterised by literature data for (i) their relative ability to scavenge the ABTS^·+ radical cation, reflected by the so-called trolox equivalent antioxidant capacity (TEAC) value, (ii) their relative rate of oxidation by a range of free radicals, or (iii) their capacity to inhibit lipid peroxidation in multilamellar liposomes, leading to a decrease in formation of thiobarbituric acid reactive substances (TBARS). All these antioxidant values for radical scavenging action correlate quantitatively with computer-calculated ionisation potentials of the carotenoids. These correlations are observed both when the ionisation potential is calculated as the negative of the energy of the highest occupied molecular orbital (-E(HOMO)) of the molecule, or as the relative change in heat of formation (ΔΔHF) upon the one-electron oxidation of the carotenoids.

The calculations provide a theoretical assay able to characterise the intrinsic electron donating capacity of an antioxidant, in hydrophilic, hydrophobic or artificial membrane environment.     

Combining optical and electrical impedance techniques for quantitative measurement of confluence in MDCK-I cell cultures

A new method combining optical and electrical impedance measurements is described that enables submicroscopic cell movements to be monitored. The cells are grown on small gold electrodes that are transparent to light. This modified electrical cell-substrate impedance sensor (ECIS) allows simultaneous microscopic recording of both growth and motility, thus enabling cell confluence on the electrodes to be systematically correlated to the impedance in regular time intervals of seconds and for extended periods of time. Furthermore, the technique provides an independent measure of monolayer cell densities that we compare to calculated values from a theoretical model. We have followed the attachment and spreading behavior of epithelial Madin-Darby canine kidney strain I (MDCK-I) cell cultures on microelectrodes for up to 40 h. The studies reveal a high degree of correlation between the measured resistance at 4 kHz and the corresponding cell confluence in 4- to 6-h intervals with typical linear cross-correlation factors of r equaling approximately 0.9. In summary, the impedance measured with the ECIS technique provides a good quantitative measure of cell confluence.     

The space-use strategy of plants with different growth forms,in a field experiment with manipulated nutrients and light

The biomass allocation pattern in plants is known to depend on the below and above-ground resource availabilities. In a herbaceous multi-species stand, it can be expected that the effects of nutrient and light availability on plants’ general space-use strategy are fundamentally different. We hypothesized that nutrient status alters the amount of biomass produced per unit canopy volume (biomass density), but not so much the biomass vertical distribution pattern. Changes in light availability, in contrast, should affect the vertical distribution pattern of biomass but not biomass density. We were also interested in whether the effect of resource manipulation on a plant’s space-use strategy depends on its basic morphological characteristics (growth form). The results from a four-year permanent plot experiment in a species-rich grassland, with fertilization and additional illumination from mirrors applied to 40 × 40 cm plots, showed that our main hypothesis was correct. Fertilization significantly affected biomass density above as well as below-ground, while additional illumination generally did not. Light addition altered the vertical distribution pattern of above-ground biomass, which remained unaffected by the fertilizer treatment.     

Horseradish peroxidase-catalyzed oligomerization of ferulic acid on a template of a tyrosine-containing tripeptide

Ferulic acid (FA) is an abundantly present phenolic constituent of plant cell walls. Kinetically controlled incubation of FA and the tripeptide Gly-Tyr-Gly (GYG) with horseradish peroxidase and H2O2 yielded a range of new cross-linked products. Two predominant series of hetero-oligomers of FA linked by dehydrogenation to the peptidyl tyrosine were characterized by electrospray ionization (tandem) mass spectrometry. One series comprises GYG coupled with 4-7 FA moieties linked by dehydrogenation, of which one is decarboxylated. In the second series 4-9 FA moieties linked by dehydrogenation, of which two are decarboxylated, are coupled to the tripeptide. A third series comprises three hetero-oligomers in which the peptidyl tyrosine is linked to 1-3 FA moieties of which none is decarboxylated. Two mechanisms for the formation of the FA-Tyr oligomers that result from the dualistic, concentration-dependent chemistry of FA and their possible role in the regulation of plant cell wall tissue growth are presented.     

Riboflavin-photosensitized changes in aqueous solutions of alginate. Rheological studies

Interactions between photoexcited riboflavin (RF), promoted by irradiation in the range of 310-800 nm, and alginate have been studied in air equilibrated aqueous solutions with the aid of rheological methods. Light irradiation of RF causes under aerobic conditions fragmentation of alginate and a decrease in the shear viscosity and other rheological parameters of its solutions. The decrease is most pronounced in concentrated polymer solutions. The photochemical degradation of alginate is inhibited in the presence of the quenchers/scavengers d-mannitol, glutathione, potassium iodide, and sodium azide and in excess oxygen. The addition of thiourea to alginate-RF solutions leads to enhanced degradation of the polymer. Significant shear-thinning effects and deviations from the Cox-Merz rule are observed at higher polymer concentrations.