Biphasic fluence-response curves for phytochrome-mediated kalanchoë seed germination : sensitization by gibberellic Acid

The fluence-response curves for the effect of two red pulses separated by 24 hours on the germination of Kalanchoe blossfeldiana Poelln. cv Vesuv seeds, incubated on gibberellic acid (GA₃) are biphasic for suboptimal concentrations. The response in the low fluence range corresponds with a classical red/far-red reversible phytochrome mediated reaction. GA₃ induces an additional response in the very low fluence range, which is also phytochrome mediated. The sensitivity to phytochrome-far-red absorbing form (Pfr), however, is increased about 20,000-fold, so that even far-red fluences become saturating. Both in the very low and low fluence response range, the maximal responses induced by saturating fluences are modulated by the GA₃ concentration. GA₃ having no direct influence on the phytochrome phototransformations, alters the Pfr requirement and determines the responding seed population fraction in the very low and low fluence range. The effet of GA₃ appears to be on the transduction chain of the phytochrome signal.     

Distribution,length and weight of roots in young plantations ofEucalyptus grandis W. Hill ex Maiden irrigated with recycled water

Summary The root systems ofEucalyptus grandis W. Hill ex Maiden, irrigated with recycled municipal effluent at two sites in north-western Victoria, Australia, were studied by excavation and coring. Trees at Robinvale were four years-old and were irrigated using micro-sprays that covered only 70% of the ground surface area, whereas at Mildura, effuent was uniformly was uniformly applied to six years-old trees by flood and sprinkler irrigation. At Mildura where roots were excavated from a 2.80×2.80×1.20 m block of soil, a total root length of 1193 m.m⁻² and a total root weight of 3.1 kg m⁻² were estimated in the top metre. For roots >1 mm diameter, 77% of intercepts were at 0–30 cm, whereas only 50% were in the 50–100 cm soil horizon. At both sites where roots in the top 30 cm were studied by coring, the vertical distributions of root intercepts, length and weight were similar. Root length was greatest in the 0–10 cm soil horizon at both sites, and intercepts of roots <1 mm diameter comprised 73% and 81% of all roots at Mildura and Robinvale respectively. Roots <1 mm diameter contributed 85% of total length at both sites, but only 19% and 21% of total weight at Mildura and Robinvale respectively. The horizontal distribution of roots differed at the two sites. With uniform application of effuent at Mildura, root intercepts and length were concentrated in the centre of the irrigation bay, but at Robinvale, the concentration occurred closer to the tree row due mainly to the different method of irrigation. Root weight at both sites was highest within 50 cm of the tree row. Root densities of 0.11 to 0.57 cm cm⁻³ were estimated in the two plantations; these were similar to root densities measured inPinus radiata D. Don plantations up to 46 months old, but were considerably lower than those estimated for pastures. The implications of the results for the management of irrigated plantations of eucalypts are discussed.     

Gas exchange characteristics of the sorghum-striga host-parasite association

Gas exchange characteristics are reported for both members of the sorghum-Striga host-parasite association. Both Striga hermonthica (Del.) Benth and Striga asiatica (L.) Kuntze had transpiration rates considerably in excess of those of sorghum (Sorghum bicolor (L.) Moench, cv CSH1). Stomatal conductance in both Striga spp. showed little response to periods of darkness and moderate water stress. Low rates of net CO₂ fixation and high rates of dark respiration led to no net daily (24 hours) C gain, and Striga would appear to be reliant on its host for photosynthate. Infection of sorghum plants with either S. hermonthica or S. asiatica reduced host photosynthetic capacity. Infected sorghum plants were also more prone to water stress, but reduced rates of CO₂ fixation could not be accounted for in terms of lower stomatal conductance. Lower stomatal conductances were associated with an increase in water use efficiency (WUE) in uninfected sorghum; however, Striga-infected sorghum plants had lower WUE than those of uninfected plants. We suggest that Striga exerts a specific effect on processes affecting C acquisition in sorghum leaves. The water relations of S. hermonthica and S. asiatica are not characteristic of plants growing in semiarid environments and are more likely to reflect the nature of the parasitic life-style. Despite transfer of water and solutes from host to parasite, the reduction in C fixation observed in infected sorghum plants appears to be the major determinant of growth reductions observed in sorghum supporting Striga.     

Inhibition by bromoestrogens of the effects of estradiol on apomorphine-induced climbing behavior

The chronic administration of estrogens to mice or rats will result in antidopaminergic effects. Apomorphine-induced climbing behavior in mice, the result of direct stimulation of dopamine receptors in the striatal and mesolimbic regions, is a simple animal model for examining these antidopaminergic effects of estrogens. Bromoestrogens, inhibitors of catechol estrogen formation, have been utilized in order to examine the role of estrogen metabolism in dopaminergic antagonism. Mice were pretreated for 3 days with 2-bromoestradiol, 4-bromoestradiol, or 2,4-dibromoestradiol dibenzoates alone or in combination with estradiol benzoate prior to apomorphine administration. The haloestrogens did not alter the climbing-induced responses elicited by apomorphine, whereas estradiol benzoate clearly attentuated the actions of apomorphine. Furthermore, the bromoestradiol dibenzoates were effective in reversing the effects of estradiol benzoate when the two steroids (estradiol benzoate and a bromoestrogen dibenzoate) were administered simultaneously during pretreatment. Thus, the bromoestrogens are able to inhibit the antidopaminergic effects of estradiol exhibited in the apomorphine-induced mouse climbing model.     

Expression of retroviral vectors in transgenic mice obtained by embryo infection.

Pre-implantation embryos were infected with the retroviral vector MMCV-neo, which carries the neomycin resistance (neo) gene and the v-myc gene. Three transgenic substrains (M-TKneo 1-3) were derived which stably transmit a single intact copy of the vector. In all of the substrains, expression of the neo gene from the internal thymidine kinase (TK) promoter was detected, with two of the substrains expressing the gene in all tissues analysed. In the third substrain, the vector had integrated on the X chromosome and neo expression varied between different tissues. A second series of transgenic mice were obtained with the retroviral vector SAX, in which the human adenosine deaminase cDNA (ADA) is under the control of an internal SV40 promoter. Four substrains (M-SAX 1-4) were analysed; however, no expression of the ADA cDNA was detected. In all mice, no expression was found of the genes under the control of the viral 5' long terminal repeats (LTRs). In the M-TKneo substrains the vector was hypomethylated irrespective of its expression whereas in the M-SAX mice the vector was hypermethylated. These results demonstrate for the first time that the TK promoter can apparently express a gene in all tissues of adult mice and that retroviral vectors with internal promoters may provide an alternative to DNA injection for the efficient expression of genes in transgenic mice.     

Environmental factors influencing the vertical migration of planktonic rotifers in a hypereutrophic tarn

The diel vertical migration of planktonic rotifers in a small, hypereutrophic tarn was investigated on four occasions in 1983. When the tarn was isothermal the rotifers were distributed throughout the water column. After stratification, the rotifers were confined to the top 1–2 m of oxygenated water. On all four dates the rotifers were aggregated at specific depths in the water column. On some occasions, the pattern of aggregation changed as the animals performed distinct diurnal migrations. Keratella cochlearis, K. quadrata and Polyarthra vulgaris usually followed the reverse migrations of the phytoplankton. In contrast, the movements of Anuraeopsis fissa were less pronounced and were associated with variations in the depth of the oxycline.     

Definition of the surface antigens of Mycobacterium malmoense and use in studying the etiology of a form of mycobacteriosis

Mycobacterium malmoense is the latest of a roster of atypical mycobacteria implicated in pulmonary infections. Yet it lacks recognizable phenotypic features to allow its ready identification. Some 23 clinical isolates of M. malmoense were examined for homologous seroagglutination reactions and characteristic surface antigens. One group showed concordant agglutination interreactions and an identical spectrum of glycolipids and are regarded as M. malmoense sensu stricto. The glycolipids are of the newly found, trehalose-containing lipooligosaccharide class. De-O-acylation followed by high-pressure liquid chromatography revealed one major and several minor oligosaccharides. Partial acidic cleavage to release glycosidically linked trehalose, alpha-mannosidase digestion to demonstrate the presence of a non-reducing-end mannobiose, perdeuteriomethylation, partial acid hydrolysis, reduction, and O ethylation, combined with 1H nuclear magnetic resonance and electron impact and fast-atom bombardment mass spectrometry revealed the structure of the major oligosaccharide as alpha-D-Manp-(1----3) -alpha-D-Manp-(1----[2-alpha-L-Rhap-(1--]4--3)-alpha-L-Rh ap- (1----3)-alpha-D-Glcp-(1----1)-alpha-D-Glcp, in which two of the 2-alpha-L-Rhap residues are O methylated at C-3. (Man, mannose; Rha, rhamnose; Glc, glucose; p, pyranosyl). The structures of the minor oligosaccharides were also determined; they differ at the distal nonreducing end. The dominant oligosaccharide was acylated by octanoate, 2-methyleicosanoate, and 2,4-dimethylpentacosanoate to yield the major species-specific surface antigen of M. malmoense, which we regard as the most characteristic feature of the pathogen.     

Recruitment of naphthalene dissimilatory enzymes for the oxidation of 1,4-dichloronaphthalene to 3,6-dichlorosalicylate, a precursor for the herbicide dicamba.

Pseudomonas putida expresses plasmid-encoded enzymes and regulatory proteins for the dissimilation of naphthalene through salicylate and the alpha-keto acid pathway. A strain of P. putida (NAH:Tn5/G67) defective in salicylate hydroxylase (nahG) was assessed for its ability to oxidize 1,4-dichloronaphthalene. Washed cell suspensions were shown to accumulate 3,6-dichlorosalicylate, which, after further chemical treatment, yields the herbicide dicamba (3,6-dichloro-2-methoxybenzoate). However, the rate of dichlorosalicylate formation from dichloronaphthalene was less than 1% of the rate of salicylate formation from unsubstituted naphthalene.     

Affinity labeling of delta-opiate receptors using [D-Ala2,Leu5,Cys6]enkephalin. Covalent attachment via thiol-disulfide exchange

[D-Ala2,Leu5,Cys6]Enkephalin (DALCE) is a synthetic enkephalin analog which contains a sulfhydryl group. DALCE binds with high affinity to delta-receptors, with moderate affinity to mu-receptors, and with negligible affinity to kappa-receptors. Pretreatment of rat brain membranes with DALCE resulted in concentration-dependent loss of delta-binding sites. Using 2 nM [3H][D-Pen2,D-Pen5]enkephalin (where Pen represents penicillamine) to label delta-sites, 50% loss of sites occurred at about 3 microM DALCE. Loss of sites was not reversed by subsequent incubation in buffer containing 250 mM NaCl and 100 microM guanyl-5'-yl imidodiphosphate (Gpp(NH)p), conditions which cause dissociation of opiate agonists. By contrast, the enkephalin analogs [D-Ala2,D-Leu5]enkephalin, [D-Ser2,Leu5,Thr6]enkephalin, [D-Pen2,D-Pen5]enkephalin, and [D-Ala2,D-Leu5,Lys6]enkephalin were readily dissociated by NaCl and Gpp(NH)p, producing negligible loss at 3 microM. This suggests that DALCE binds covalently to the receptors. Pretreatment of membranes with the reducing agents dithiothreitol and beta-mercaptoethanol had no effect on opiate binding. Thus, loss of sites required both specific recognition by opiate receptors and a thiol group. The irreversible effect of DALCE was completely selective for delta-receptors. Pretreatment with DALCE had no effect on binding of ligands to mu- or kappa-receptors. The effect of DALCE on delta-binding was: 1) markedly attenuated by inclusion of dithiothreitol in the preincubation buffer, 2) partially reversed by subsequent incubation with dithiothreitol, 3) slightly enhanced when converted to the disulfide-linked dimer, and 4) prevented by blocking the DALCE sulfhydryl group with N-ethylmaleimide or iodoacetamide. These results indicate that DALCE binds covalently to delta-receptors by forming a disulfide bond with a sulfhydryl group in the binding site. The mechanism may involve a thiol-disulfide exchange reaction.