ACUTE TOXICITY OF SOME BLUEGREEN ALGAE TO THE PROTOZOAN PARAMECIUM CAUDATUM1

Four species of bluegreen algae were tested for possible effect on the protozoan Paramecium caudatum Ehrenberg. Toxicity was demonstrated using lyophilized cells of Fischerella epiphytica Ghose and Gloeotrichia echinulata (Smith) Richter. Nostoc linckia (Roth) Bornet & Thuret failed to show any effects when lyophilized but became toxic when sonified. Anabaena flos-aquae (Lyngb.) Bréb. was nontoxic in all tests. G. echinulata was lethal at 0.1 mg·ml^?1 which is comparable to the toxic concentration of Aphanizomenon flos-aquae (L.) Ralfs reported for microcrustaceans.     

Genetic Analysis of Trichocyst Discharge of the Wild Stocks of Paramecium tetraurelia*

SYNOPSIS. Aberrant discharge of trichocysts in response to picric acid occurs in 8 of the 28 wild stocks of Paramecium tetraurelia. There are at least 4 distinguishable phenotypes: nondischarge, stocks 139, 163, 169, and 242; temperature-sensitive nondischarge, stock 126; leaky nondischarge, stock 203; and a clonally unstable phenotype, stocks 146 and 148. From each of these stocks a single recessive gene causing nondischarge has been isolated by backcrosses to stock 51. The original stocks 126, 146, and 148 possess other genes which affect the extracted genes. The copper resistance locus is ～ 10 centiMorgans from nd169 and nd242, but none of the other nondischarge genes are linked to 6 marker loci. The genes nd169 and nd242 are only 0.5 centiMorgans apart making them the closest known pair of loci in P. tetraurelia. The genes nd126 and nd242 are distinguishable alleles at the same locus and the genes nd146 and nd148 are apparently identical alleles. The large number of loci involved in producing a similar phenotype in different stocks supports the idea that mutation is much more important than gene flow in this highly inbreeding species.     

Motility Events of Trichocyst Insertion in Paramecium tetraurelia*

SYNOPSIS. Following electroshock-induced extrusion of its inserted trichocysts, Paramecium tetraurelia rapidly begins replacement of the population of lost organelles. Light microscopy of the cortical insertion of new trichocysts reveals a series of characteristic motility activities. An uninserted trichocyst in the cyclotic flow of the cell appears to be “captured” and removed to the noncyclotic, subcortical regions. The trichocyst then makes a series of saltatory motions which apparently serve to transport it to the cortex, with proper orientation (tip first) for insertion. Trichocyst saltations end with either cortical insertion of the organelle, or return to cyclosis. If the trichocyst is inserted, it makes a series of unique pivoting movements around the motionless tip. This form of motility, termed “wobble,” continues for a short period of time. After cessation of wobble, the insertion of the trichocyst is apparently complete, since no further motility is observed. With the aid of these observations it was possible to identify saltatory motility as the means for transporting trichocysts to the cortex for insertion, and also to observe a motility of unknown significance (wobble) apparently associated with the process of cortical insertion.     

Comparison of Singlet and Doublet Paramecium tetraurelia: DNA Content,Protein Content and the Cell Cycle*

SYNOPSIS Doublet Paramecium tetraurelia contain either a single macronucleus which is substantially larger than that in a singlet cell, or 2 smaller macronuclei. Doublets have approximately twice the DNA content and twice the total protein content of singlets. The cell cycle of doublets is 164% as long as that of singlets, but the relative position of the macronuclear DNA synthesis period within the cell cycle is the same as in singlets. The DNA content of doublets is regulated so that differences in the number of macronuclei do not produce corresponding changes in DNA content; bimacronucleate doublets have only 27% more DNA than unimacronucleate doublets.     

Die stoffwechselphysiologischen Beziehungen zwischen Paramecium bursaria Ehrbg. und Chlorella spec. in der Paramecium bursaria-Symbiose

Zusammenfassung Infektionsexperimente algenfreier Paramecium bursaria mit aus diesen isolierten und unter Stickstoffmangel-Bedingungen vorkultivierten Algen deuten darauf hin, daß die Versorgung der endosymbiontischen Algen mit stickstoffhaltigen Verbindungen durch ihren Wirt in einem zu gutem Wachstum und Vermehrung der Alge ausreichendem Maße möglich ist. Die Bedeutung dieser stoffwechselphysiologischen Beziehung für die Symbiosepartner wird diskutiert.Die Vergiftung der Photosynthese der endosymbiontischen Chlorella durch 3-(3,4-Dichlorphenyl)-1,1-dimethylharnstoff (DCMU) führt in grünen Paramecium bursaria durch Beeinflussung des Kohlenstoff-Stoffwechsels zu einer Entkoppelung des symbiontischen steady state-Systems und damit zur Auflösung der Symbiose. Eine ausreichende heterotrophe Ernährung der Alge durch das Paramecium ist in der Symbiose offenbar nicht möglich.Die Anwendung von 3-(3,4-Dichlorphenyl)-1,1-dimethylharnstoff (DCMU) kann als neue Methode zur Züchtung algenfreier Paramecium bursaria dienen.

---

The metabolic interactions between Paramecium bursaria Ehrbg. and Chlorella spec. in the Paramecium bursaria-symbiosisI. The nitrogen and the carbon metabolism

Symbiotic Chlorellae have been isolated from Paramecium bursaria Ehrbg. and cultivated under conditions of nitrogen deficiency. Reinfection of Chlorella-free Paramecium bursaria with these nitrogen-deficient algae resulted in a complete regeneration and multiplication of the algae within the host cells. The endosymbiotic algal cells of the Paramecium bursaria-symbiosis can be supplied by their host with nitrogen.The inhibition of photosynthesis by 3-(3,4-Dichlorophenyl)-1,1-dimethylurea (DCMU) leads in green Paramecium bursaria to a breakdown of the symbiotic steady state-system resulting in a loss of algal cells. Obviously the endosymbiotic algae cannot be fed heterotrophically by their host to such an extent that a stable symbiosis is maintained.The application of 3-(3,4-Dichlorophenyl)-1,1-dimethylurea (DCMU) can be used as a new method for culturing Chlorella-free Paramecium bursaria.

     

Parafusin and calcium-induced signal transduction in exocytosis

Summary The phosphoglycoprotein parafusin is a member of the phosphoglucomutase superfamily and has been shown, both via biochemical and localization studies, to be associated with the Ca²⁺-dependent regulated exocytosis process inParamecium tetraurelia. Stimulation of exocytosis in this cell leads to a Ca²⁺-dependent glucosylation of parafusin accompanied by its dissociation from the secretory vesicles and from cell membrane docking sites. These events are blocked in the presence of extracellular Mg²⁺ in wild-type cells and in either Ca²⁺ or Mg²⁺ in a temperature-sensitive mutant, nd9, stimulated at the nonpermissive temperature. Furthermore, laser scanning confocal localization studies with antibodies to parafusin whole protein versus antibody made to a specific peptide (insertion 2) show different localization patterns. While insertion-2 antibodies only label the organelles previously shown to have parafusin associated with them, i.e., cell membrane fusion (docking) sites and secretory vesicles, antibodies to whole protein outline in addition the alveolar sacs (subsurface cisterns) which are Ca²⁺ storage compartments in this cell. This may indicate tht other members of the phosphoglucomutase superfamily which interact specifically with this compartment are present inP. tetraurelia.     

Indispensable function of the germ nucleus for postconjugational stomatogenesis in Paramecium caudatum: Evidence against its genic function shown by hypohaploid micronuclei

It is known that the germinal micronucleus at the stages of gametogenesis and/or fertilization has an indispensable function for the postconjugational development of oral apparatus (stomatogenesis) in Paramecium caudatum. To determine whether this function is due to some specific genes in the micronucleus, postconjugational stomatogenesis was examined in the conjugation of haploid and hypohaploid cells. Haploid clones were obtained by conjugation between amicronucleate cells and diploid micronucleate cells. After conjugation between these haploid clones or between the haploid clones and amicronucleate clones, we succeeded in obtaining hypohaploid clones that have various types of nullisomic micronuclei. If a few genes in the micronucleus control postconjugational stomatogenesis, some hypohaploid micronuclei should undergo stomatogenesis normally, but others should not. In the present work, however, almost all the hypohaploid micronuclei developed the oral apparatus and formed food vacuoles. We can apparently rule out the possibility that a few specific genes of the micronucleus are required for postconjugational stomatogenesis in Paramecium caudatum, unless selection operates to retain the chromosomes with the essential gene(s). Dev. Genet. 23:142–150, 1998. © 1998 Wiley-Liss, Inc.     

Micronucleus‐Specific Bacterium Holospora elegans Irreversibly Enhances Stress Gene Expression of the Host Paramecium caudatum

ABSTRACT. The bacterium Holospora is an endonuclear symbiont of the ciliate Paramecium. Previously, we reported that paramecia bearing the macronuclear‐specific symbiont Holospora obtusa survived better than symbiont‐free paramecia, even under high temperatures unsuitable for growth. The paramecia with symbionts expressed high levels of hsp70 mRNAs even at 25 °C, a usual growth temperature. We report herein that paramecia bearing the micronuclear‐specific symbiont Holospora elegans also acquire the heat‐shock resistance. Even after the removal of the bacteria from the hosts by treatment with penicillin, the resulting aposymbiotic paramecia nevertheless maintained their heat shock‐resistant nature for over 1 yr. Like symbiotic paramecia, these aposymbiotic paramecia also expressed high levels of both hsp60 and hsp70 mRNAs even at 25 °C. Moreover, analysis by fluorescent in situ hybridization with a probe specific for Holospora 16S rRNA revealed that the 16S rRNA of H. elegans was expressed around the nucleoli of the macronucleus in the aposymbiotic cells. This result suggests the possible transfer of Holospora genomic DNA from the micronucleus into the macronucleus in symbiotic paramecia. Perhaps this exogenous DNA could trigger the aposymbiotic paramecia to induce a stress response, inducing higher expression of Hsp60 and Hsp70, and thus conferring heat‐shock resistance.     

Phylogenetic position of endosymbiotic green algae in Paramecium bursaria Ehrenberg from Japan

Endosymbiotic green algae of Japanese Paramecium bursaria were phylogenetically analyzed based on DNA sequences from the ribosomal DNA operon (18S rDNA, ITS1, 5.8S rDNA, and ITS2). Phylogenetic trees constructed using 18S rDNA sequences showed that the symbionts belong to the Chlorella sensu stricto (Trebouxiophyceae) group. They are genetically closer to the C. vulgaris Beijerinck group than to C. kessleri Fott et Nováková as proposed previously. Branching order in C. vulgaris group was unresolved in 18S rDNA trees. Compared heterogeneities of 18S rDNA, ITS1, 5.8S r, and ITS2 among symbionts and two Chlorella species, indicated that the ITS2 region (and probably also ITS1) is better able to resolve phylogenetic problems in such closely related taxa. All six symbiotic sequences obtained here (approximately 4000-bp sequences of 18S rDNA, ITS1, 5.8S rDNA, and ITS2) were completely identical in each, strongly suggesting a common origin.