Reversion of Streptococcal Protoplasts

The reversion of protoplasts of Streptococcus faecium was examined by light and scanning electron microscopy. Regularly shaped streptococci emerged from the narrow region of large, oval protoplasts.     

Concentration of Enteroviruses on Membrane Filters

Enteroviruses can be made to adsorb or to pass through membrane filters by manipulation of the suspending medium. Salts facilitate virus adsorption, but membrane-coating components (MCC) interfere. Because cells release MCC into the culture medium during viral growth, MCC must be removed before virus can be adsorbed to membranes. Adsorbed virus can be eluted with diluents containing MCC (cell extracts or serum) or agents that reduce surface tension (sodium lauryl sulfate). By membrane adsorption and elution, enteroviruses can be readily concentrated and quantitatively recovered from crude virus harvests.     

Reversion to the Streptococcal State of Enterococcal Protoplasts, Spheroplasts, and L-Forms

A method is described for predetermining whether lysozyme-damaged enterococci grow as either the parent strain or as L-forms. Organisms treated with lysozyme grew as L-forms on media solidified with low concentrations of agar, or the damaged cells grew as streptococci on media solidified with high concentrations of gelatin. After induction, some of the L-forms reverted to the parent strain, but most did not during three routine subcultures. Continued spontaneous reversion occurred through approximately 30 subcultures after induction. However, subsequent progeny did not revert, even when subjected to conditions such as the gelatin medium which strongly favors growth in the streptococcal phase.     

Pressure Effects on Membrane Potentials of Mesophyll Cell Protoplasts and Epidermal Cell Protoplasts of Commelina communis L.

Pantoja, O. and Willmer, C. M. 1986. Pressure effects on membranepotentials of mesophyll cell protoplasts and epidermal cellprotoplasts of Commelina communis L.—J. exp. Bot. 37:315–320. Membrane potentials of epidermal cell protoplasts and mesophyllcell protopiasts of Comnelina communis were measured when theprotoplasts were immobilized in a suction micropipette. Whenzero suction was employed, membrane potentials of both protoplasttypes were near to zero. As suction pressure was increased,membrane potentials became increasingly more negative with gradientsof 14·3 mV/kPa and 10·5 mV/kPa for mesophyll cellprotoplasts and epidermal cell protoplasts, respectively. Theplasma membrane is stretched when suction pressure is appliedto protoplasts and it is considered that this simulates cellturgor pressure which is associated with negative membrane potentialsof intact cells. The results help to explain why some investigatorsobtain positive membrane potentials for protoplasts while othersobtain negative values. The results also indicate that considerablecaution is needed in the interpretation of ion flux data whenprotoplasts are used. Key words: Commelina communis, membrane potentials, pressure, protoplasts     

Influence of Coliform Source on Evaluation of Membrane Filters

Four brands of membrane filters were examined for total and fecal coliform recovery performance by two experimental approaches. Using diluted EC broth cultures of water samples, Johns-Manville filters were superior to Sartorius filters for fecal coliform but equivalent for total coliform recovery. Using river water samples, Johns-Manville filters were superior to Sartorius filters for total coliform but equivalent for fecal coliform recovery. No differences were observed between Johns-Manville and Millipore or Millipore and Sartorius filters for total or fecal coliform recoveries using either approach, nor was any difference observed between Millipore and Gelman filters for fecal coliform recovery from river water samples. These results indicate that the source of the coliform bacteria has an important influence on the conclusions of membrane filter evaluation studies.     

Isolation and Counting of Athiorhodaceae with Membrane Filters

The number, type, and distribution of Athiorhodaceae in two Central Pennsylvania artificial lakes were investigated with an anaerobic modification of the membrane filter technique.     

Scanning Electron Microscopy of Microbial Cells on Membrane Filters

Scanning electron micrographs of a Pseudomonas species, Staphylococcus aureus, and Bacillus subtilis on two membrane filtration systems are compared.     

Selective Mycobacteriophage Isolation with Membrane Filters

A simple technique is described whereby mycobacteriophages are selectively isolated by adsorption on mycobacteria layered within membrane filter sandwiches before standard enrichment procedures.     

Efficient Filtration and Sizing of Viruses with Membrane Filters

Untreated membrane filters retain viruses by adsorption, as well as by physical restriction which occurs when the pore diameter of the filter is smaller than that of the virus particle. As originally recommended by Elford, membranes had to be pretreated with proteinaceous material to preclude virus adsorption. However, coating materials that prevent adsorption of certain viruses do not necessarily prevent adsorption of other viruses. In contrast to proteins, salts enhance virus adsorption. Viruses treated with sodium lauryl sulfate to reduce the surface tension, or purified viruses in distilled water, are not adsorbed to membranes. A procedure is recommended by which viruses may be passed through membranes with a porosity twice the diameter of the virus. Such filtrates, which contain 50 to 100% of the initial virus concentration, should be used for sizing viruses by subsequent filtration through smaller pores. The determination of virus size would then be based on the major population of particles in the virus suspension. In the past, as little as 0.1 to 0.001% of the initial virus population was the basis for size determination, because more than 99.9% of the virus was often lost by adsorption to membranes during the clarifying procedures.     

Auxin-Induced Expansion Growth of Cells and Protoplasts of Yeast

Using an auxin-responsive mutant of Sacchairomyces ellipsodeus, expansion growth of cells caused by auxin was studied especially in comparison with that of protoplasts.

• 1 Indole-3-acetic acid induced detectable cell expansion growth in 3 hours in a buffered simple solution where no cell division occurred.
• 2 The auxin-induced expansion growth was inhibited by an antiauxin, trans-cinnamic acid.
• 3 Actinomycin D, chloramphenicol and cycloheximide inhibited the auxin-induced cell expansion growth.
• 4 Protoplasts did not expand in response to auxin under the condition where intact cells did.
• 5 The stability of protoplasts was not changed by the low auxin concentration (20 mg/1) which induced cell expansion.
• 6 High concentrations (100–1000 mg/1) of auxin caused protoplasts to burst even under an osmotically stable condition.

     

Growth of Amoebae and Flagellates on Bacteria Deposited on Filters

Abstract Artificial bacterial biofilms were formed by making microwave-irradiated, dual-radioisotope-labelled Vibrio bacteria adhere to 0.4 μm pore size filters with albumin. The rate of release of ³H from thymidine label in these bacteria into the surrounding seawater when protozoa were incubated with the biofilm indicated the predator's grazing rate, and the rate of accumulation of ¹⁴C in the predators from leucine label in the bacteria indicated the assimilation rate of the protozoa. The amoeba Vanella septentrionalis consumed about 60% of the available bacteria between the 5th and 15th days of incubation with a gross growth efficiency of 22 ± 6%, compared with about 75% consumption at 29 ± 8% efficiency for the surface-feeding flagellate Caecitellus parvulus, and about 55% consumption at 16 ± 5% efficiency for the suspension-feeding flagellate Pteridomonas danica. As a result of their grazing and metabolism these protozoa regenerated about 70–85% of the nutrients present in their food and released these nutrients in the immediate vicinity of the bacterial biofilm. The biomass of the amoeba Vanella was calculated to be 166 pg protein cell⁻¹ during maximum growth and 93 pg protein cell⁻¹ in the stationary phase. Received: 3 August 1998; Accepted: 20 November 1998     

甘蔗和烟草叶原生质体分离期间的膜损伤及有关酶活性变化

甘蔗和烟草叶原生质体分离期间的膜损伤及有关酶活性变化何若天,覃伟,李任强（广西农业大学实验中心,南宁５３０００５）关键词：原生质体,超氧阴离子自由基（Ｏ_２~－）,膜损伤,甘蔗,烟草植物原生质体分离期间,所用细胞壁降解酶和高渗介质等对细胞生理有深刻影响...     

Energetics of Streptococcal Growth Inhibition by Hydrostatic Pressure

Growth of Streptococcus faecalis in complex media with various fuel sources appeared to be limited by the rate of supply of adenosine-5′ -triphosphate (ATP) at 1 atm and also under 408 atm of hydrostatic pressure. Growth under pressure was energetically inefficient, as indicated by an average cell yield for exponentially growing cultures of only 10.7 g (dry weight) per mol of ATP produced compared with a 1-atm value of 15.6. Use of ATP for pressure-volume work or for turnover of protein, peptidoglycan, or stable ribonucleic acid (RNA) did not appear to be significant causes of growth inefficiency under pressure. In addition, there did not seem to be an increased ATP requirement for ion uptake because cells growing at 408 atm had significantly lower internal K⁺ levels than did those growing at 1 atm. Pressure did stimulate the membrane adenosine triphosphatase (ATPase) or S. faecalis at ATP concentrations greater than 0.5 mM. Intracellular ATP levels were found to vary during the culture cycle from about 2.5 μmol/ml of cytoplasmic water for lag-phase or stationary-phase cells to maxima for exponentially growing cells of about 7.5 μmol/ml at 1 atm and 5.5 μmol/ml at 408 atm. N,N′-dicyclohexylcarbodiimide at a 10 μM concentration improved growth efficiency under pressure, as did Mg²⁺ or Ca²⁺ ions at 50 mM concentration. These agents also enhanced ATP pooling, and it seemed that at least part of the growth inefficiency under pressure was due to increased ATPase activity. In all, it appeared that S. faecalis growing under pressure has somewhat reduced ATP supply but significantly increased demand and that the inhibitory effects of pressure can be interpreted largely in terms of ATP supply and demand.     

K+ Channels in the Plasma Membrane of Lily Pollen Protoplasts

Using the patch-clamp technique K⁺ channels could be observed in the plasma membrane of protoplasts from pollen grains of Lilium longiflorum. With depolarizing membrane potentials the open probability of the different K⁺ channels increased. Two K⁺ channel populations occurring occasionally had a single channel conductance of 120 pS and 42 pS, respectively. The most often observed K⁺ channel had a single channel conductance of 19 pS which showed an increase of channel activity with increasing free cytoplasmic Ca²⁺ concentration. This channel population might be involved in the pathway of endogenous transcellular K⁺ currents which are activated during pollen tube tip extension.     

Relationship Between Yield of Protoplasts and Growth Phase in Saccharomyces

A number of generations must occur in the presence of glucose before daughter cells of stationary or old cultures become amenable to protoplast formation by the action of snail enzyme.     

Factors Influencing the Growth and Division of Pea Mesophyll Protoplasts

High yields of viable protoplasts were produced from pea leaves provided that only leaves of the same age were used in each preparation. The conditions under which the pea plants were grown and the age of the plants were also important. The protoplasts were cultured in a medium supplemented with 1 mg/1 2iP and 1 mg/1 2,4-D. They were able to regenerate cell walls within two days. After 5 days cell divisions were apparent and sustained divisions led to callus formation. Special emphasis has been given in this paper to the choice of leaf material for protoplast isolation.