Cell Surface Saccharides in Three Phytomonas Species Differing in Host Specificity

ABSTRACT. Cell surface carbohydrates of three phytoflagellates, Phytomonas francai. Phytomonas serpens and Phytomonas sp. from different hosts including cassava, coreid insect Phthia picta and the milkweed plant Euphorbia hyssopifolia, respectively, were analysed by agglutination assays employing a battery of highly purified lectins with affinity for receptor molecules containing N-acetylglucosamine (d-GlcNAc), N-acetylgalactosamine (D-GalNAc), galactose, mannose-like (D-Man-like) residues and fucose, and by binding assay using radiolabeled [¹²⁵I]-wheat germ agglutinin (WGA) and fluorescent WGA lectin, as well as glycosidases of known sugar specificity, Escherichia coli K with mannose-affinity fimbrial lectin was also used as an agglutination probe. In general, the presence of D-GlcNAc. D-GalNAc and D-Man-like residues was detected in the phytomonads' plasma membrane. These sugar moieties were confirmed in whole cell hydrolysates as assessed by gas-liquid chromatography (GLC) which in addition, also showed the presence of galactose and xylose. However, marked differences in cell surface carbohydrate structures were observed. Wheat germ agglutinin, which binds to sialic acid and/or d-GlcNAc-containing residues, shows selective agglutinin activities for P. francai and Phytomonas sp., while Bandeiraea simplicifolia II agglutinin (which recognizes d-GlcNAc units) specifically bound to Phytomonas sp. Helix pomatia agglutinin which binds to D-GalNAc-containing residues reacted preferentially with Phytomonas sp. and P. serpens. Con A, which recognizes D-Man-like receptors, agglutinates all the phytomonads; however, the higher interaction was observed with Phytomonas sp. P. francai was selectively agglutinated in the presence of E. coli fimbrial lectin. Fluorescence WGA binding was significantly decreased by N-acetylglucosaminidase activities and the cell agglutination was not altered by neuraminidase treatment, suggesting the presence of an exposed D-GlcNAc moiety on the P. francai and Phytomonas sp. surfaces. Binding studies with [¹²⁵I]-WGA essentially confirmed the fluorescence WGA binding and agglutination assays.     

Changes in Cell Surface Anionogenic Groups Induced by Propranolol in Herpetomonas muscarum muscarum

The effects of propranolol (10⁻³ mM) on the surface anionic groups of Herpetomonas muscarum muscarum were analysed by cell electrophoresis, by ultrastructural cytochemistry and by identification of sialic acids using paper chromatography. Differentiation of H. muscarum muscarum induced by propranolol treatment caused a significant increase in the net negative surface charge. Binding of cationized ferritin (CF) and colloidal iron hydroxide particles was observed at the cell surface of both untreated and propranolol-treated cells. In cells incubated in the presence of the drug the CF particles were distributed in all membrane regions. However, there were small areas where the particles were absent. In H. muscarum muscarum exposed to propranolol the density of residues of sialic acid per cell was higher, and the agglutinating activity with Sendai virus was more intense. However, the pattern of sialic acid, characterized by the presence of N-acetylneuraminic acid derivative, was not modified upon cell interaction with the drug. Treatment of both control and propranolol-treated protozoa with neuraminidase significantly reduced the surface charge. These findings suggest that sialic acid residues are the major anionogenic groups exposed on the surface of H. muscarum muscarum .     

Strain Differences in the Ultrasonic Behavior of Rats (Rattus Norvegicus)

Wistar rat pups are generally more "vocal" than Lister pups;the ultrasonic calls that they produce are usually louder andemitted at a greater rate than those of Lister pups of the sameage and studied under similar conditions of isolation or handling.These differences in ultrasonic behavior are apparently notrelated to differences in the development of homiothermy whichfollows a similar course in both strains. The calls of infantsof the two strains do not show any marked differences in durationor frequency pattern that are consistent over age or treatmentcondition, but the Wistar pups appear to show greater changeswith age than the Lister pups in certain parameters of the calls,while the latter strain show greater changes with treatmentcondition. Similar strain differences in the rate and intensity of callingwere seen during aggressive encounters between adult males,but these cannot be related to differences in aggressive behavior.The strains differ in the duration of both the long "22 kHz"calls (Lister greater than Wistar) and the shorter "50 kHz"calls (Wistar greater than Lister), as well as in the minimumfrequency of these shorter calls (lower in Wistar than in Listerrats).     

Programmed cell death during development of cowpea (Vigna unguiculata (L.) Walp.) seed coat

The seed coat develops primarily from maternal tissues and comprises multiple cell layers at maturity, providing a metabolically dynamic interface between the developing embryo and the environment during embryogenesis, dormancy and germination of seeds. Seed coat development involves dramatic cellular changes, and the aim of this research was to investigate the role of programmed cell death (PCD) events during the development of seed coats of cowpea [Vigna unguiculata (L.) Walp.]. We demonstrate that cells of the developing cowpea seed coats undergo a programme of autolytic cell death, detected as cellular morphological changes in nuclei, mitochondria, chloroplasts and vacuoles, DNA fragmentation and oligonucleosome accumulation in the cytoplasm, and loss of membrane viability. We show for the first time that classes 6 and 8 caspase‐like enzymes are active during seed coat development, and that these activities may be compartmentalized by translocation between vacuoles and cytoplasm during PCD events.     

Reinstatement of Phyllanthus retroflexus Brade (Phyllanthaceae)

Phyllanthus retroflexus Brade is reinstated. It and the very similar P. subemarginatus Müll. Arg. are described, illustrated, and their relationships are discussed, based on a morphological analysis of herbarium specimens. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 78–81.     

Membrane-Associated Polysaccharides Composition, Nutritional Requirements and Cell Differentiation in Herpetomonas roitmani: Influence of the Endosymbiont

Herpetomonas roitmani , a trypanosomatid containing a bacterial endosymbiont, was cured by high doses of chloramphenicol. Wild-type and cured flagellates were compared as to polysaccharide composition, nutritional requirements and cellular differentiation. Fucose (18.0%), xylose (15.7%), mannose (38.9%), galactose (10.8%), glucose (16.4%) and inositol (< 1.0%) were identified as polysaccharide components of cured H. roitmani as assessed by gas-liquid chromatography. However, the wild-type strain displayed a markedly different sugar profile, in that xylose was absent and inositol preferentially synthesized, whereas the other monosaccharide components remained unchanged. Variations in nutritional pattern also occurred between both strains. The bacterial endosymbiont seems to provide the flagellates with nutritional factors, including usual amino acids, vitamins, purine (as adenine) and hemin. The process of cell differentiation was also significantly influenced by the endosymbiont. Opisthomastigote forms predominate (72.0%) in cured as compared with wild-type H. roitmani (37.0%).     

Cell Surface Carbohydrate Differences in Wild and Mutant Strains of Crithidia fasciculata1

Wild type Crithidia fasciculata and three drug-resistant mutant strains that have shown “flagellar adherence” were studied as to their ability to agglutinate with lectins specific for receptor molecules containing N-acetyl glucosamine, N-acetyl galactosamine, galactose, mannose-like residues, fucose, and sialic acid. Escherichia coli with mannose-sensitive fimbriae was also used as an agglutination probe. The presence of D-GalNAc, D-Gal, and mannose-like residues was detected in the wild strain. Generally, in the mutants, residues of these sugar units were present in greater concentrations when compared to the wild type strain. β-Galactosidase treatment showed that β-D-Galp units are exposed on the cell membrane. All types of cell agglutination including flagellum-flagellum (F-F), flagellum-soma (F-S), and soma-soma (S-S) were observed when lectins were used; however, with E. coli only the F-F type of cell agglutination was observed with the wild type strain and the TFR^R1 mutant. All types of agglutination were observed with the other two mutants.     

Evolutionary tendencies in some annual species of Brotnus (Bromus L. sect. Genea Dum. (Poaceae))

A multidisciplinary investigation of the eight annual taxa of Bromus sect. Genea throughout their total ranges has brought a fresh insight into their evolutionary pathways. These species, mainly in southwest Asia and the Mediterranean, are still actively in the process of speciation and their study illuminates evolutionary processes in annual grasses as a whole. Following a discussion of Genea features, a list of ancestral and derived characters in the group is presented. The recent emergence of a multiplicity of niches is probably the reason for the rapid emergence of many new kinds of plants in sect. Genea.