X-Ray Micro-Analysis of Cells and Cell Compartments of A triplex spongiosa: I. LEAVES

Atriplex spongiosa was grown in hydroponic culture with additionsof 0, 200, 400 and 600 mM NaCl. Frozen, hydrated cells and cellcompartments of young and mature leaf tissue were analysed byX-ray micro-analysis. Evidence for low K + Na and Cl content,and high K selectivity in the bundle sheath cytoplasm was obtainedfrom data on X-ray count ratios and on total X-ray counts. Vacuolesof the major cell types of the mature leaf had either high Kor high Na and Cl contents when grown in the absence or presenceof NaCl. Comparison of K, Na and Cl content of different cell types inthe mature leaf showed gradients in selectivity for K. relativeto Na between the bundle sheath cells and the bladder cells.In the young expanding leaves salt was sequestered in the numeroussalt bladders on the leaf surface, while the cytoplasm and developingvacuoles of undifferentiated cells contained largely K and littleNa or Cl. The results support general views on the compartmentation ofsalt in plants cells in relation to osmotic or saline stress. Key words: Atriplex spongiosa, X-ray analysis, Salinity, Compartmentation, Leaf     

Localization of Glyoxylate Cycle Enzymes in Glyoxysomes in Euglena*

SYNOPSIS. We demonstrated previously microbodies in Euglena gracilis grown in the dark on 2-carbon substrates. We have now established in Euglena the particulate nature of enzymes known in other organisms to be localized in microbodies (glyoxysomes and leaf peroxisomes). On a linear sucrose gradient the glyoxylate cycle enzymes band together at a nigner equilibrium density (1.20 g/cm3) than mitochondrial marker enzymes (1.17 g/cm3), establishing the existence in Euglena of glyoxysomes similar to those of higher plants. Glyoxylate (hydroxypyruvate) reductase and, under certain conditions, also glycolate dehydrogenase co-band with the glyoxylate cycle enzymes, suggesting that Euglena glyoxysomes, like those of higher plants, may contain peroxisomal-type enzymes. Catalase, an enzyme characteristic of microbodies from a variety of sources, was not detected in Euglena.     

Failure of Neuraminidase to unmask Histocompatibility Antigens on Trophoblast

DURING outbred pregnancy the mother is exposed to genetically foreign tissue because the offspring inherits transplantation antigens from the father. The survival of the foetus is ensured by the intervention of the trophoblast which does not express transplantation antigens between mother and foetus: mouse trophoblast is not rejected even when transplanted into immune recipients^1,3. The mechanism of this failure to express histocompatibility antigens is not understood^1–4, but Kirby et al. have suggested that the extracellular fibrinoid surrounding trophoblast cells is involved^5,6. Currie has suggested that the thick sialomucinous glycocalyx of the trophoblast cell might “mask” the histocompatibility antigens on the trophoblast^7,8 and has demonstrated that neuraminidase unmasked these antigens⁸. Our experiments, however, show that trophoblast incubated with neuraminidase cannot sensitize allogeneic mice to donor histocompatibility antigens. Furthermore, pretreatment of trophoblastic implants with neuraminidase did not interfere with their proliferation and growth in highly immune allogeneic recipients.     

Serial Passage of Trypanosoma lewisi in the Heterologous Mouse Host. I. Development in Calorically-restricted Hosts.

SYNOPSIS. Two isolates ("A" and "B") of Trypanosoma lewisi from the same rat stock source were serially transferred in calorically-restricted mice supplemented daily with normal rat serum. The "A" strain was transferred consecutively through 300 mice over a period of more than 3 years and was voluntarily discontinued. The "B" strain died out spontaneously after 43 consecutive passages in mice. The developmental histories of these 2 isolates were analyzed and compared with respect to duration of the parasitemic period, interval to the next subsequent passage in mice, day of death of each host animal, proportion of host animals that died, intensity of parasitemia in mouse tail blood, interval required for development of the observed maximal parasitemia, and duration of maximal parasitemia.
"A" appeared to have become progressively adapted to the mouse as judged by a decrease in parasitemic period with successive transfer associated with a progressive increase in trypanosome population, and declines in the interval required for development of the observed maximal parasitemia and in the duration of this maximal response. There did not appear to be any correlation of the percentage of animals that died with any other factor. The "B" strain did not appear to have adapted itself to mice as judged by the foregoing criteria.     

Nucleic Acids of Entamoeba histolytica

This review will concentrate on certain aspects of the nucleic acids of Entamoeba histolytica. Utilization and synthesis of purines and pyrimidines will initially be briefly discussed, e.g. salvage vs. de novo pathways, uptake studies and recognition of at least 4 transport loci. Data will be presented which show that the distribution and synthesis of RNA (to a lesser extent DNA) in the nucleus is basically the opposite one finds in other eukaryotes, viz. most RNA (ribosomal?) is synthesized (or accumulates) in the peripheral chromatin (functional equivalent of nucleolus?). The DNA is distributed and synthesized primarily throughout the nucleus. It is usually so dispersed that it will not stain with e.g. the standard Feulgen technique, unless the DNA condenses around the endosome (not a nucleolar equivalent) prior to nuclear division. Isolation of rRNA was difficult due, in part, to potent and difficult to inhibit RNase(s), some of which are apparently intimately bound to ribosomal subunits. The 25S (1.3 kDa), 178 (0.8 kDa) and 58 rRNA were recovered after isolation with a high salt SDS-DEP technique. This is the only procedure which enables us to obtain high yields of 258 rRNA: guanidine or guanidinium which permits isolation of intact functional mRNA results in isolation of small amounts of 28 RNA relative to 178 RNA. The 258 RNA is “nicked” (apparently during nuclear processing) and dissociates readily into 1 78 (0.7 kDa) and 168 (0.6 kDa) species, and a more rigidly bound 5.88 species. A small amount of “unnicked” 258 RNA was recovered with guanidine. Two DNA-dcpendent RNA polymerases (I and II) with a pronounced preference for denatured DNA as template were eluted from DEAE-Sephadex in reverse order of what occurs in other eukaryotes, except Physarum polycephalum. This conclusion was based on salt optima and alpha-amanitin sensitivity studies. Initial characterization of DNA isolated with a procedure capable of isolating > 100-kbp Leishmania DNA showed that undigested DNA migrates as a broad band between markers 6 and 24 kbp. The persistent recovery of such a “band” by us and Perez-Mutul et al. no larger than ca. 24 kbp (with the exception of >48 kbp DNA isolated by Hernandez et al. using an in situ lysis technique which did not include a proteinase) may be due to nicks introduced during isolation; or, perhaps much of the amebal DNA exists in vivo as gene sized fragments. However, preliminary data generated using orthogonal pulse-field agarose gel electrophoresis do suggest that amebal DNA may be in small chromosomes.     

Functional Constraints on the Evolution of Larval Forms of Marine Invertebrates: Experimental and Comparative Evidence

Marine invertebrate larvae are well known for their distinctivebody shapes and elaborate patterns of ciliation. In this studyI take a physically based approach to investigate the functionalconsequences of variations in body shape and patterns of ciliation.With experimental models I demonstrate that shape as well assurface area contributes to drag of larval forms. Based on flowfields around larvae tethered in still water and flowing waterI argue that drag, which acts as a partial tether, may influencehow water is processed and food is captured by cilia. With mechanicalmodels of cilia I show that placement of cilia on the surfacescan influence the effectiveness with which water is moved andthe steepness of the velocity gradient through the ciliary layer.These models indicate that placement of cilia on ridges, atextreme anterior ends, and at extreme posterior ends of larvalbodies increases the volume of water moved per ciliary strokerelative to placement of cilia on a flat surface. A comparativesurvey of46 larval forms indicates that distributions of bodyshape and patterns of ciliation reflect functional requirementsof swimming and feeding by larvae. The experimental and comparativeapproaches together suggest functional constraints on the evolutionof larval forms which may lead to convergence in patterns ofciliation and conservation of larval forms within taxa.     

Nitrogen Fixation by Subterranean Clover at Varying Stages of Nodule Dehydration: II. EFFICIENCY OF NITROGENASE FUNCTIONING

Changes in nitrogenase activity (C₂H₂ reduction and H₂ production),nodulated root respiration and the efficiency of nitrogenasefunctioning were measured in response to progressive dehydrationof nodules on intact well-watered plants of subterranean clover(Trifolium subterraneum L.) cv. Seaton Park. The nodulated rootsof vegetative plants grown to the 14-leaf stage were incubatedin a gas exchange system through which a continuous dry airstreamwas passed over an 8 d period. The root tips were immersed inan N-free nutrient solution during this time so that water andion uptake was unimpeded. The decline in nodulated root respirationresulting from nodule drying was associated with a continualreduction in respiration coupled to nitrogenase activity. Asnodule water potential (_nod) decreased, the proportion of totalnodulated root respiration which was nitrogenase-linked declinedfrom 50% (day 1) to 33% (day 8). This was accompanied by a 79%reduction in specific nitrogenase activity (from 3.79 to 0.81umol C₂H₄ g^–1 nodule dry weight min^–1). Nodule dehydrationalso induced a decline in hydrogen (H₂) production in air. Therelative decline in hydrogen production exceeded that of acetylenereduction activity and this resulted in an increase in the relativeefficiency of nitrogenase functioning. However, the carbon costof nitrogenase activity progressively increased above 2.0 molCO₂ respired per mol C₂H₄ reduced as rood decreased below –0.4to –0.5 MPa. Consecutive measurements of the rates ofhydrogen evolution, ¹⁵N₂ fixation and acetylene reduction activityon intact unstressed plants resulted in a C₂H₄/N₂ conversionfactor of 4.08 and an electron balance of 1.08. These resultsindicated that the pre-decline rate of acetylene reduction activitymeasured in a flow-through system provided a valid measure ofthe total electron flux through nitrogenase. Key words: Subterranean clover, dehydration, efficiency, nitrogenase activity     

Epiphytic diatoms in two freshwater maritime Antarctic lakes

SUMMARY.

• 1 An ecological study of two small maritime Antarctic lakes on Signy Island, South Orkney Islands, was undertaken from January 1986 to March 1987. Analysis of diatom counts from the lakes provided examples of oligotrophic and mesotrophic ecosystems.
• 2 A diverse community of 104 epiphytic taxa was identified. Twenty-eight taxa had a percentage abundance greater than 1% in both lakes. Distinctive dominant taxa were identified from each lake.
• 3 A variety of ordination techniques was performed on the abundance data and a principal components analysis demonstrated differences in the diatom assemblages between the two lakes. Clear separations of sites and species were evident between the lakes, and accounted for the greatest percentage variance.
• 4 Species composition varied with depth within each lake and was also important in influencing changes in assemblage composition between sites.
• 5 A redundancy analysis indicated that species composition was correlated to concentrations of nitrogen and phosphorus but the total variance accounted for by the four physical and chemical factors measured was low (24%).