Characterization of tetranucleotide microsatellite loci and development and validation of multiplex reactions for the study of right whale species (genus Eubalaena)

A North Atlantic right whale (Eubalaena glacialis) genomic library was developed and screened with a (GATA)₈ probe to identify tetranucleotide microsatellite loci. Sixteen characterized loci were polymorphic in North Atlantic and/or South Atlantic (Eubalaena australis) right whales, 12 being polymorphic in E. glacialis, and 15 in E. australis. Fourteen of these were combined with 21 other previously identified loci for a suite of 35 loci which can be used to increase resolution of genetic analyses of these species. Multiplex reactions were developed for genotyping samples at these loci, providing a method that is rapid, reliable and cost‐effective.     

Mineralization of soil nitrogen in three forest communities from the New England region of New South Wales

Nitrogen (N) mineralization rates and the temperature response patterns of mineral N production in surface (0–7.6 cm) soils were compared in laboratory incubation studies based on disturbed, composite samples. Seasonal variation in the field levels of mineral N, and mineralization potential of intact (7.6 × 5.6 cm diameter) soil cores, were also investigated. Ammonification proceeded rapidly in each soil. Nitrification did not occur in grassy forest (GF) soil but was active in both layered forest (LF) and mossy forest (MF) soils, especially the former. Total mineral N production was greatest in MF and least in LF. Ammonification in disturbed samples was maximal at 50°C in all three soils with a secondary peak at 10°C in LF soil. Nitrification in LF and MF soils was most rapid at 25°C. Several species of ammonifying bacteria with different temperature optima were isolated, indicating that the process of ammonification is a composite of the activities of a variety of decomposer microbes. Mean field levels of mineral N and NH₄–N throughout the year were greatest in MF and least in LF. Seasonal fluctuations in NH₄–N were evident, concentrations being universally low in mid-winter (about 1.5 μgg^-1), increasing to a maximum in late summer (about 5 μg g^-1 in LF: 16–18 μg g^-1 in GF and MF). Field levels of NO₃–N were more constant and never more than 5 μg g^-1 in any community. Both total mineralization and ammonification in intact cores were greatest in MF and least in LF while nitrification was greatest in LF and almost negligible in GF, thus confirming the results obtained with disturbed samples. The potential for mineralization was large in mid-winter when the amount of mineral N was very low, and small in late summer when field levels were higher: this is interpreted as indicating that seasonal climatic factors regulate the availability of substrates for decomposers. Spatial variability in field levels of mineral N and mineral N production in the laboratory was evidenced by significant ‘sampling site’ effects in each community: however, at the sampling intensity used, the presence of bark mounds around Eucalyptus saligna trees could not be shown to affect these attributes. The inability of GF soil to nitrify when incubated in the laboratory could not be ascribed to a high C/N ratio, low pH, lack of substrate ammonium, or a low population of autotrophic nitrifying bacteria. No attempt was made to investigate the presence of allelopathic nitrification inhibitors. No evidence was obtained to support the view that nitrification is atypical of climax communities in situ. The most productive forest (LF) had the greatest capacity to nitrify and the least productive community (GF) the smallest capacity to do so.     

Ion Transport and the Effects of Acetic Acid in White Clover: I. PHOSPHATE ABSORPTION

Dunlop, J., Knighton, M. V. and White, D. W. R. 1988. Ion transportand the effects of acetic acid in white clover. I. Phosphateabsorption.—J. exp. Bot. 39: 79–88. The effect of acetic acid on phosphate absorption by white clovertissue has been examined. At 1·0 mol m³ acetic acid phosphateabsorption by roots of intact plants was stimulated by 36% (P< 0.001). At 5.0 and 10 mol m^–3 acetic acid there wasmarked inhibition of absorption by both suspension culturesof cells and the roots of intact plants. The inhibition waspH dependent with decreasing pH causing increased inhibition.Acetic acid caused changes in the membrane electropotential(E) with concentrations of 2·0 mol m^–3 or lesscausing persistent polarization whereas at 5·0 mol m^–3and higher concentrations the polarization was followed by agreater depolarization. Intracellular pH as measured by thefluorescence of fluorescein was lowered by acetic acid. Calculationsindicate that for white clover roots the proton motive force(pmf) appears to provide sufficient energy for phosphate absorption.It is proposed that acetic acid influences phosphate absorptionthrough its dependence on proton cotransport and that changesin J E affect the rate of phosphate absorption because of thedependence of the pmf on E. Key words: Phosphate absorption, intracellular pH, acetic acid, proton motive force, Trifolium repens, membrane electropotential     

Ion Transport and the Effects of Acetic Acid in White Clover: II. POTASSIUM ABSORPTION

Dunlop, J., Knighton, M. V. and White, D. W. R. 1988. Ion transportand the effects of acetic acid in white clover. II. Potassiumabsorption.—J. exp. Bot. 39: 89–96. Acetic acid stimulated K⁺ influx into cells of white cloverin suspension culture and net K⁺ influx by roots of intact whiteclover plants. At concentrations of acetic acid up to 2·0mol m^–3 the stimulation continued unabated for at least2 h. However, at higher concentrations the rate of absorptiondeclined to near zero values after 2 h. When acetic acid wasremoved from the solution there was a net efflux of K⁺. Thestimulation was pH dependent with a maximum at pH 5·0.There was maximum stimulation at an acetic acid concentrationof 2·0 mol m^–3 Net H⁺ efflux was reduced by 1·0 mol m^–3 aceticacid. When Rb⁺ uptake and H⁺ efflux were measured at a rangeof RbCl concentrations Rb⁺ uptake increased with concentrationwhereas H⁺ efflux was maximum in the absence of Rb⁺ (and K⁺)and decreased as RbCl concentration was increased Acetic acid caused a hyperpolarization of the membrane electricalpotential difference (E) of about 25 mV. In 1·0 mol m^–3acetic acid the hyperpolarization persisted for at least 2 hwhereas at 10 mol m^–3 d E subsequently depolarized tovalues around –80 mV. With a slight lag, the time courseof the stimulation of the rate of K⁺ absorption followed thepolarization and depolarization of E. These results imply thatthe linkage between K⁺ and H⁺ movements is probably throughE. Key words: Proton efflux, membrane electrical potential difference, Trifolium repens     

Interspecific Hybridization of Trifolium repens with T. hybridum Using In Ovulo Embryo and Embryo Culture

In ovulo embryo culture followed by culture of excised immatureembryos produced interspecific hybrids between Trifolium repensL. (white clover) and autotetraploid T. hybridum L. (alsikeclover). Ovules containing hybrid embryos were excised 12–14 dafter pollination and cultured on Nitsch (1951) medium supplementedwith 15% young cucumber juice for 5–6 d. Embryos weresubsequently excised and transferred to hormone-free EG medium,a medium suitable for the culture of immature embryos. A total of 118 hybrid seedlings were obtained from 1978 reciprocalpollinations. All seedlings produced showed various chlorophylldeficiencies, either totally albino or albino with green sectors.Transmission electron microscope studies were carried out toinvestigate plastid development in embryos and seedlings. Someembryos produced only callus. Plants were regenerated from sevencalli. Two semi-albino plants survived transfer to soil, andone plant produced flowers. Backcrosses to T. repens producedone green plant. Hybridity is supported by analysis of morphological characters,karyotype and the gel electrophoretic separation of leaf isozymes. Pollen irradiated with 40 Gy of gamma rays was also used forpollinations. Results indicate that in certain cases ionizingradiation might be useful in overcoming hybrid inviability. Trifolium repens, Trifolium hybridum, clover, interspecific hybridization, in ovulo embryo culture, irradiation     

The freezing characteristics of cauliflower curd

Cultivar differences in frost resistance and the heritable nature of resistance were demonstrated using seedling cauliflower plants. Such cultivar differences were not however expressed in the curd. Selection for frost resistance in cauliflower should therefore use whole plant screening techniques. Curd material when frozen as isolated florets, supercooled over the range – 1°C to – 12°C and the mean freezing point of all curds tested was -6°C to -7.25°C (overall mean -6.44°C). Curd florets which supercooled but did not freeze were completely undamaged, whereas freezing always led to cell damage and death observed as water-soaking of the floret surface and measured using an electrical conductivity method. The large range of freezing points measured suggests a range of active ice nucleators either on or within the florets. When curds were frozen intact the ability of florets to supercool was severely restricted which was attributed to the seeding of freezing by the internal growth of ice crystals. A crop protection strategy needs to identify and control or modify warm temperature nucleators in cauliflower curd.