Axenic Paramecium caudatum. I. Mass Culture and Structure*

SYNOPSIS. To establish and grow Paramecium caudatum in mass axenic culture the culture medium of Soldo, Godoy & van Wagtendonk was modified by substituting phosphatidylethanolamine (PE) for TEM-4T and by a 10-fold increase in folic acid. Population densities of 4000 to 6000 cells/ml and a generation time of 20–26 h are regularly obtained. Optimal growth is obtained with PE-stigmasterol ratios between 40:1 to 400:1. Cells from 1-day-old axenic cultures have many lipid bodies aggregated in clumps (which disappear in 2 to 3 days) as well as foci of rough endoplasmic reticulum bordered by dictyosomes. The latter suggests a very active metabolism. Crystalline sheets found in both food vacuoles and lysosomes presumably play a role in digestion. Axenically grown cells also have abundant Golgi bodies (dictyosomes) and by late log phase become filled with lysosomes.     

Membrane Dynamics of the Contractile Vacuole Complex of Paramecium1

ABSTRACT. Membrane dynamics of the contractile vacuole complex of Paramecium were investigated using conventional electron microscopy of cells so that the vacuoles were serial-sectioned longitudinally and transversely. During systole, vacuolar membrane collapses first into flattened cisternae which undergo further modification into a mass of interconnected small membrane tubules. These tubules retain their connections with the radiating microtubular ribbons; consequently they are found only in the poleward hemisphere. Permanent connections between ampullae and the collapsed vacuole membrane could not be verified nor was a sphincter-like mechanism for closing such a junction observed. Membranes of the ampullae and the collecting canals also collapse to varying extents into arrays of tubules that remain bound to microtubular ribbons during diastole. Thus vacuole, ampullae, and collecting canal membranes all assume tubular forms when internal volume is at a minimum. Having failed to observe a microfilamentous encasement of the vacuole, we suggest that an alternative mechanism for the “contractile” function should be sought. One such is based on fluid volume increase and fluid flow within transiently interconnected tubular membrane systems that cycle between a tubular and a planar membrane form as internal volume is periodically increased and reduced. The driving force for this mechanism might best be sought in the molecular structure of the membranes of the contractile vacuole complex.     

Monoclonal Antibody to a Bacterial Endonuclear Symbiont Holospora Cross Reacts with Proteins of Contractile Vacuole Radial Canals of Paramecium Species

ABSTRACT A monoclonal antibody (mAb) IR-2-1 was raised against a 67-kDa protein purified from the macronucleus-specific bacterial symbiont Holospora obtusa of Paramecium caudatum. The mAb was found to react with two bands (31 and 67-kDa) on gels of H. obtusa. Indirect immunofluorescence microscopy showed that these antigens were distributed inside the cells. However, unexpectedly, this mAb also cross reacted with the radial arms of the contractile vacuole in P. caudatum, P. tetraurelia, P. multimicronucleatum, P. jenningsi and P. bursaria as well as with their cytoplasm. Immunoelectron microscopy showed that the antigens were located on the decorated spongiome of the radial arms. In immunoblots, mAb IR-2-1 reacted with a band of 67 kDa in all Paramecium species examined. However, no band appeared in the immunoblot of isolated macronuclei of H. obtusa-free P. caudatum and no label was seen in the nuclear matrix of the macronucleus of air-dried P. caudatum. These results suggest that the 67-kDa antigen found in H. obtusa was not imported from the host macronucleus and the same antigen in the host contractile vacuoles and cytoplasm were not derived from the symbiont. These results also showed that an epitope on the decorated spongiome of the Paramecium species is shared by its bacterial symbiont. In contrast to the decorated tubule-specific mAb, DS-1, the antigens for IR-2-1 appeared to be loosely membrane bound as they were lost in paraformaldehyde fixed and acetone permeabilized Paramecium. Supplementary key words. Contractile vacuole complexes, Holospora obtusa, monoclonal antibody, Paramecium.     

The probing behaviour of nymphs of Vanduzeea arquata and Enchenopa binotata (Homoptera: Membracidae) on host and non-host plants

Abstract. 1. Nymphs of Vanduzeea arquata Say have been found to be more host-specific in nature and to show a higher degree of selectivity in host discrimination experiments than nymphs of Enchenopa binotata (Say), It was hypothesized that this differential selectivity would be reflected in the probing behaviour of individuals placed on twigs of host and non-host plants. Probing behaviour was examined by direct observation of nymphs and by sectioning and staining the probed plant tissues.
2. All nymphs probed readily and for extended periods on both host and non-host twigs. E.binotuta nymphs showed no consistent differences in probing behaviour on hosts versus non-hosts, but V.atquuta nymphs were more likely to withdraw their stylets within 60 s when on non-host twigs and produced honeydew only when on their host species. V.urquatu nymphs reached the phloem sieve elements only when on host twigs and broke many cells in peripheral plant tissue layers while probing. E.binotata nymphs broke few cells and often reached the phloem of non-host as well as host plants.
3. Nymphs of V.arquata always reject non-host plants, apparently in the course of probing and prior to encountering the phloem sap. Chemical compounds released from ruptured parenchyma cells may act as probing stimulants or inhibitors. E.binotura nymphs often feed on non-host plants in a non-choice situation; their preferential settling on host twigs in discrimination experiments may reflect a tendency to abandon non-host twigs more readily than host twigs.     

Endogene Jahresperiodik der Blattbewegungen zweier Oxalis-Arten

Under natural light-dark changes at the Arctic Circle special behaviour was observed in the leaf movement of Oxalis regnellii and Oxalis acetosella: (1) Advancing in the beginning of leaf rising. (2) Prolonging of the duration of day position. (3) Increasing positive phase-andle differences. (4) Prolonging of the duratin of the daily moving-time. From January to April all these features were running back. The winter behaviour has also been shown in artificial light-dark changes (12:12), when plants were taken in the trial before mid winter. The reactions did not occur if the plants were put into artificial light conditions after mid winter. Criterions for an endogenous annual periodicity and its possible annual Zeitgebers under natural light-dark changes are discussed. The positive correlation between the length of day position and the length of day-time in the summer respectively the length of night-time in the winter, as well as positive phase-angle difference in spring and in autumn/winter, are judged as a criterion for a second oscillator.