INFLUENCE OF TRICHOME LENGTH ON LINEAR TRANSLATION IN OSCILLATORIA PRINCEPS (CYANOPHYCEAE)1

Motile trichomes of Oscillatoria princeps Voucher were examined to determine the relationship between trichome length and the ability to glide through hardened or viscous media. A minimum length for movement in solid media was found to be 0.1 mm in 0.5% agar (w/v).Viscous media tests revealed that coordinated movements through methyl cellulose media were not seen in trichomes 15–40 μ long at the viscosity employed. The minimum length motility increases with the viscosity of the medium and the gliding rates observed are greater in longer trihomes. These findings are discussed in relation to trichome surface area and the hypothesis that the motility system is restricted to lateral surfaces of the trichome.     

Soil stabilization by a prokaryotic desert crust: Implications for Precambrian land biota

A cyanophyte dominated mat, desert crust, forms the ground cover in areas measuring hundreds of square meters in Utah and smaller patches in Colorado. The algal mat shows stromatolitic features such as sediment trapping and accretion, a convoluted surface, and polygonal cracking. Sand and clay particles are immobilized by a dense network of filaments of the two dominating cyanophyte species,Microcoleus vaginatus andM. chthonoplastes, which secrete sheaths to which particles adhere. These microorganisms can tolerate long periods of desiccation and are capable of instant reactivation and migration following wetting. Migration occurs in two events: 1. immediately following wetting of dry mat, trichomes are mechanically expelled from the sheath as it swells during rehydration, and 2. subsequently, trichomes begin a self-propelled gliding motility which is accompanied by further production of sheath. The maximum distance traveled on solid agar by trichomes ofMicrocoleus vaginatus during a 12 hour period of light was 4.8 cm. This corresponds to approximately 500 times the length of the fastest trichome, and provides a measure of the potential for spreading of the mat in nature via the motility of the trichomes.Dehydration resistence of the sheath modifies the extracellular environment of the trichomes and enables their transition to dormancy. Following prolonged wetting and evaporative drying of the mat in the laboratory, a smooth wafer-like crust is formed by the sheaths ofMicrocleus trichomes that have migrated to the surface. Calcium carbonate precipitates among the algal filaments under experimental conditions, indicating a potential for mat lithification and fossilization in the form of a caliche crust. It is suggested that limestones containing tubular microfossils may, in part, be of such an origin.The formation of mature Precambrian soils may be attributable to soil accretion, stabilization, and biogenic modification by blue-green algal land mats similar to desert crust.     

GLIDING MOTILITY IN THE BLUE-GREEN ALGA OSCILLATORIA PRINCEPS1

Gliding is an active movement displayed by a microorganism in contact with a solid substrate where there is no evidence of a motility organelle or of a conformational change in the organism. Gliding may be accompanied by rotations, reversals, flectional activity, and mucilage sheath production, as well as linear translation. Previous explanations of the mechanism responsible did not consider all these aspects of behavior. The gliding behavior and ultrastructure of the blue-green alga Oscillatoria princeps Vaucher were examined. O. princeps has a maximum observed gliding rate of 11.1 μm/sec. The trichomes can glide in either longitudinal direction following rapid and occasionally frequent reversals. Right-handed trichome rotation was always observed, which means that any surface point on these trichomes traces a 60-deg right-handed helix. A mucilage sheath envelopes the moving trichomes. The rate of gliding was reduced by viscous substrates, extreme pH, lysozyme, DNP, and cyanide, while sustained darkness had no inhibitory effect. Ultrastructurally, the cell wall is composed of an L-1 layer which is 10 nm thick and often ill-defined. The L-2 layer which is outside this is 200 nm thick and participates in septum formation. The L-3 layer is outside the L-2 and is continuous over the trichome surface. The L-4 “membrane” lies outside the L-3 layer. Grazing surface sections and freeze-etch replicas show a parallel and tight array of 6–9 nm wide continuous fibrils in the cell wall on the surface of the distinctive L-2 layer. Isolated wall fragments were tightly coiled inside out with the fibrils on the inside. The angle of orientation for the fibrils was to the right in a helix with a pitch of 60 deg. O. animalis, a blue-green alga with a movement tracing a left-handed helix, showed a similar array of fibrils oriented in a left-handed helix with a pitch of 60 deg. It is proposed that gliding is produced by unidirectional waves of bending in the fibrils which, act against the sheath or substrate, tints displacing the trichome.     

Diel Vertical Movements of the Cyanobacterium Oscillatoria terebriformis in a Sulfide-Rich Hot Spring Microbial Mat

Oscillatoria terebriformis, a thermophilic cyanobacterium, carried out a diel vertical movement pattern in Hunter's Hot Springs, Oreg. Throughout most daylight hours, populations of O. terebriformis covered the surface of microbial mats in the hot spring outflows below an upper temperature limit of 54°C. Upon darkness trichomes moved downward by gliding motility into the substrate to a depth of 0.5 to 1.0 mm, where the population remained until dawn. At dawn the population rapidly returned to the top of the mats. Field studies with microelectrodes showed that the dense population of O. terebriformis moved each night across an oxygen-sulfide interface, entering a microenvironment which was anaerobic and reducing, a dramatic contrast to the daytime environment at the mat surface where oxygenic photosynthesis resulted in supersaturated O₂. Laboratory experiments on motility with the use of sulfide gradients produced in agar revealed a negative response to sulfide at concentrations similar to those found in the natural mats. The motility response may help explain the presence of O. terebriformis below the mat surface at night. The movement back to the surface at dawn appears to be due to a combination of phototaxis, photokinesis, and the onset of oxygenic photosynthesis which consumes sulfide.     

Community structure and pigment organisation of cyanobacteria-dominated microbial mats in Antarctica

Benthic microbial mat communities were sampled from 20 lakes, ponds and streams of the McMurdo Sound region, Antarctica. At least five distinct assemblages could be differentiated by their cyanobacterial species composition, pigment content and vertical structure. The most widely occurring freshwater communities were dominated by thin-trichome (0·5–3 µm) oscillatoriacean species that formed benthic films up to several millimetres thick. ‘Lift-off mats’ produced mucilaginous mats 1–5 cm thick at the surface and edge of certain ponds. Another group of oscillatoriacean communities was characteristic of hypersaline pond environments; these communities were dominated by species with thicker trichomes such as Oscillatoria priestleyi. Black mucilaginous layers of Nostoc commune were widely distributed in aquatic and semi-aquatic habitats. Dark brown sheath pigmentation was also characteristic of less cohesive mats and crusts dominated by Pleurocapsa, Gloeocapsa and Calothrix. High performance liquid chromatography analysis of the lipophilic pigments showed that the upper region of most of the Antarctic mats was enriched in sheath pigments (scytonemin) and/or certain carotenoids such as myxoxanthophyll and canthaxanthin. Most of the chlorophyll a (Chla), as well as phycocyanin, β-carotene and echinenone, was located in the lower strata of the mat profiles. In many of these communities most of the photosynthetic biomass occurred in a ‘deep Chla maximum’ that was well protected from short-wavelength radiation by the surface layer of light-screening pigments.     

Soil stabilization by a prokaryotic desert crust: implications for Precambrian land biota.

A cyanophyte dominated mat, desert crust, forms the ground cover in areas measuring hundreds of square meters in Utah and smaller patches in Colorado. The algal mat shows stromatolitic features such as sediment trapping and accretion, a convoluted surface, and polygonal cracking. Sand and clay particles are immobilized by a dense network of filaments of the two dominating cyanophyte species, Microcoleus vaginatus and M. chthonoplastes, which secrete sheaths to which particles adhere. These microorganisms can tolerate long periods of desiccation and are capable of instant reactivation and migration following wetting. Migration occurs in two events: 1. immediately following wetting of dry mat, trichomes are mechanically expelled from the sheath as it swells during rehydration, and 2. subsequently, trichomes begin a self-propelled gliding motility which is accompanied by further production of sheath. The maximum distance traveled on solid agar by trichomes of Microcoleus vaginatus during a 12 hour period of light was 4.8 cm. This corresponds to approximately 500 times the length of the fastest trichome, and provides a measure of the potential for spreading of the mat in nature via the motility of the trichomes. Dehydration resistence of the sheath modifies the extracellular environment of the trichomes and enables their transition to dormancy. Following prolonged wetting and evaporative drying of the mat in the laboratory, a smooth wafer-like crust is formed by the sheaths of Microcleus trichomes that have migrated to the surface. Calcium carbonate precipitates among the algal filaments under experimental conditions, indicating a potential for mat lithification and fossilization in the form of a caliche crust. It is suggested that limestones containing tubular microfossils may, in part, be of such an origin. The formation of mature Precambrian soils may be attributable to soil accretion, stabilization, and biogenic modification by blue-green algal land mats similar to desert crust.     

Ecological studies of Chloroflexis,a gliding photosynthetic bacterium

Summary Chloroflexis, a gliding, filamentous, photosynthetic bacterium, is present in the stratified algal-bacterial mats which occur in the 50°–70°C temperature range of alkaline hot spring effluents. The organism is in association with the alga in the upper, algal layer, and also forms thick, orange mats beneath the algal layer. Natural populations of Chloroflexis from these mats demonstrated light-stimulated uptake of some ¹⁴C-labelled organic compounds. Photosynthetic ¹⁴CO₂ fixation by natural samples of Chloroflexis was investigated with respect to temperature, light intensity and mat depth. Bacterial photosynthesis was determined in samples in which algae were present by use of the inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Bacterial photosynthesis was maximal at depths down to about 3 mm and then decreased rapidly to very low levels at greater depths. The greatest amounts of bacteriochlorophyll pigments were also concentrated in the top 3–4 mm of the mat. The optimum light intensity for bacterial photosynthesis (about 400 ft-c) was considerably lower than the normal summer light intensity at the surface of the mat (5000-8000 ft-c).The temperature optima for photosynthesis by the bacterial component of natural mat samples from several sites of different temperatures in a hot spring thermal gradient were determined. Temperature optima approximated the environmental temperatures, indicative of the occurrence of strains of Chloroflexis adapted to different temperatures. Although bacterial standing crop was greatest in the temperature range 50°–55°C, maximum photosynthetic efficiency was observed at about 45°C. Sulfide was stimulatory to photosynthetic ¹⁴CO₂ fixation by naturally occurring populations of Chloroflexis under field conditions. These data are consistent with the hypothesis that Chloroflexis may utilize sulfide as an electron donor for photosynthetic CO₂ reduction. However, it is also likely that Chloroflexis grows photoheterotrophically in these mats, obtaining organic compounds from algal excretory products.     

ALGAL EXCRETION AND BACTERIAL ASSIMILATION IN HOT SPRING ALGAL MATS1

Benthic algal-bacterial mats are present in the effluents of alkaline hot springs at temperatures between 50 and 73 C. The thin surface layer is composed of the unicellular blue-green alga Synechococcus lividus. Also present in the surface layer and forming thick, orange mats beneath it, are filamentous, phototrophic, gliding bacteria of the genus Chloroflexis, also capable of heterotrophic growth. The very low species diversity and the constancy of the hot spring environment, make these mats a good ecosystem for studying the transfer of nutrients from the algae to the bacteria. To determine whether the alga might supply organic materials to the bacterium, excretion by natural populations of S. lividus was studied in the field by means of short-term radioisotope experiments. Under optimal conditions for photosynthesis, between 3 and 12% of the total ¹⁴C fixed was excreted as ¹⁴C-labeled organic compounds. Variations in cell density at concentrations of S. lividus approximating those found in the mat had no effect on the percentage excretion. However, at cell densities below a threshold, level, the percentage excretion increased with diminishing cell density. Except at very low light intensities the percentage of fixed carbon excreted, was very similar for all light intensities tested. Excretion at temperatures approaching the upper limit for growth was not significantly different from the percentage excretion values observed at lower temperatures. ¹⁴C-labeled organic compounds excreted during algal photosynthesis could be subsequently assimilated by natural populations of the bacteria present in the mat.     

Electrical nature of the taxis signal in cyanobacteria

Electrical events after a light-dark stimulus were studied in the multicellular organism Phormidium uncinatum. Normally, such a stimulus causes the gliding trichome to reverse direction. By directing a large light spot on the end of a batch of trichomes and then switching it off, we achieved synchronization of the trichomes, since the "head" is much more sensitive than the "tail." The abrupt disappearance of a uniform light produced a depolarization wave which initiated at the head, as registered by externally applied electrodes. The second stimulus produced a depolarization of the opposite direction, reflecting the reorientation of the trichomes. No electrical response was observed at Ca2+ concentrations less than or equal to 10(-8) M. Factors causing oscillatory reversals, i.e., a combination of Ca2+ and A23187, or a viscous environment also abolished the electrical signal. Changes in an externally applied electrical field (4 V/cm2) had little effect on the motile behavior of P. uncinatum or Oscillatoria princeps. However, in the presence of 5 microM Ca2+-1 microM A23187, all the trichomes reversed synchronously to the anode after a change in polarity of an externally applied electrical field. We suggest that an increased Ca2+ concentration together with a change in delta psi (or delta mu H+) represents the taxis signal in cyanobacteria.     

Modeling filamentous cyanobacteria reveals the advantages of long and fast trichomes for optimizing light exposure

Cyanobacteria form a very large and diverse phylum of prokaryotes that perform oxygenic photosynthesis. Many species of cyanobacteria live colonially in long trichomes of hundreds to thousands of cells. Of the filamentous species, many are also motile, gliding along their long axis, and display photomovement, by which a trichome modulates its gliding according to the incident light. The latter has been found to play an important role in guiding the trichomes to optimal lighting conditions, which can either inhibit the cells if the incident light is too weak, or damage the cells if too strong. We have developed a computational model for gliding filamentous photophobic cyanobacteria that allows us to perform simulations on the scale of a Petri dish using over 10(5) individual trichomes. Using the model, we quantify the effectiveness of one commonly observed photomovement strategy--photophobic responses--in distributing large populations of trichomes optimally over a light field. The model predicts that the typical observed length and gliding speeds of filamentous cyanobacteria are optimal for the photophobic strategy. Therefore, our results suggest that not just photomovement but also the trichome shape itself improves the ability of the cyanobacteria to optimize their light exposure.     

A LIGHT AND ELECTRON MICROSCOPE STUDY OF INTERLAMELLAR POLYGLUCOSIDE BODIES IN OSCILLATORIA CHALYBIA

A light and electron microscopic study of interlamellar granules in Oscillatoria chalybia was made to determine their physiological nature. Oscillatoria chalybia was cultured under continuous light in media of high nitrogen content, moderate nitrogen content and low nitrogen content. Cultures growing vigorously in a medium of moderate nitrogen content were placed in darkness for an additional 96 hr. Periodic acid-Schiffs reagent tests were made on specimens from these 4 cultural conditions. Electron microscopic studies of interlamellar granules were correlated with the cytochemical tests. It is shown that diastase digestion will eliminate the PAS-positive substance and the interlamellar granules. Conclusions are that the interlamellar granules are polyglucoside in nature and that they vary in number and size with available nitrogen, light intensity and age of culture in such a way as to indicate that they are food reserves.     

Interacting effects of light,temperature, and nutrients on C-14 uptake of oscillatoria rubescens de candolle

The rate of C¹⁴O₂ uptake in a bacterized isolate of Oscillatoria rubescens indicates that within the ranges studied temperature caused the greatest variation followed by light intensity and nutrient concentration. The variation within interaction effects of light, temperature, and nutrients was higher than that within any other combination of interactions. High temperatures (25°C) shifted the light optimum of O. rubescens growing in low to moderate nutrient levels from 1950 lux to 800 lux.     

Trichome micromorphology in Turkish species of Ziziphora (Lamiaceae)

Ziziphora L. is represented by 5 species and 2 subspecies in the flora of Turkey: Z. clinopodioides, Z. capitata, Z. persica, Z. tenuior, Z. taurica subsp. taurica, Z. taurica subsp. cleonioides. It is difficult to distinguish between some Ziziphora taxa because of their morphological similarities. In this study, the leaf and calyx trichomes of Ziziphora taxa in Turkey were studied in order to assess anatomical variations that may serve as distinguishing characters. Their micromorphological features were surveyed by scanning electron microscopy (SEM) and light microscopy (LM). Trichomes on leaves and calyx can be divided into two general types: non‐glandular trichomes and glandular (secretory) trichomes. The non‐ glandular trichomes are simple, acicular or curved with cuticular micropapillae. They usually consist of one or more additional cells. The glandular trichomes are divided into two types: peltate and capitate and Ziziphora taxa can easily be distinguished by presence/absence, density and types of glandular trichomes on leaves and calyx. The peltate trichomes consist of 12 or 18 secretory head cells in a single disc; four or six central cells surrounded by eight or twelve peripheral ones. Peltate trichomes are absent on the adaxial leaf surface of Z. capitata and Z. persica. Two types of capitate trichomes are present in Ziziphora. The capitate trichomes are only absent on the calyx surface of Z. persica. In addition, the trichome micromorphology provides some support for separating the two subspecies of Z. taurica. In conclusion, Ziziphora taxa can easily be distinguished by cell number, cell shape presence/absence and density of the glandular trichomes on leaves and calyx.     

ASSOCIATION OF PLEUROCAPSA AND CALOTHRIX (CYANOPHYTA) IN A THERMAL STREAM1,2

A crustose, nodular mat, primarily composed of Calothrix sp. and Pleurocapsa sp., occurs in some alkaline hot springs of the western United States. Together, these species constituted ca. 71% of the mat biomass; the remainder was partitioned between other cyanophytes and bacteria. Their temperature growth ranges in culture were 24–50 C (Calothrix) and 30–55 C (Pleurocapsa). Both clonal cultures had maximal growth rates at 45 C (Pleurocapsa, 1.28 doublings/24 h; Calothrix 2.30). Calothrix grew at approximately twice the rate of Pleurocapsa throughout their coincidental temperature range. The relative proportions of Calothrix (42%) and Pleurocapsa (27%) were constant in a thick mat (ca. 1 mm) regardless of season; however, a thin mat (ca. 0.5 mm) contained significantly more Pleurocapsa (71%) and less Calothrix (5%). Analysis of 10 μm thick microtome sections showed that Pleurocapsa dominated the innermost region (substrate side) of the nodule mat while Calothrix abundance increased from ca. 3% in this region to 80–96% at the surface of thick mat samples (ca. 0.8–2.3 mm). Our proposed that the grazing pressure exerted by an ostracod population accounts for the continued coexistence of and the observed spatial relationships between these species appears most consistent with the observations.     

Chemokinetic Motility Responses of the Cyanobacterium Oscillatoria terebriformis

Oscillatoria terebriformis, a gliding, filamentous, thermophilic cyanobacterium, exhibited an inhibition of gliding motility upon exposure to fructose. The observed response was transient, and the duration of nonmotility was directly proportional to the concentration of fructose. Upon resumption of motility, the rate of motility was also inversely proportional to the concentration of fructose. Sulfide caused a similar response. The effect of sulfide was specific and not due to either anoxia or negative redox potential. Exposure to glucose, acetate, lactate, or mat interstitial water did not elicit any motility response.     

Function of cAMP as a Mat-Forming Factor in the Cyanobacterium Spirulina platensis

A dense suspension of Spirulina platensis trichomes aggregatedrapidly and formed a diskshaped algal mat when cAMP was added.Cyclic AMP significantly stimulated algal mat formation at concentrationsas low as 10^–7 M. Stimulation of the mat formation wasmost rapid at about 10^–5M cAMP, but higher concentrationswere not increasingly effective. Other nucleotides such as cGMP,ATP and AMP showed no stimulatory effect on algal mat formation.CCCP, an inhibitor of ATP synthesis, was found to suppress thecAMP-stimulated algal mat formation. Cyclic AMP also stimulatedrespiration and gliding movement of this cyanobacterium. (Received September 2, 1991; Accepted October 15, 1991)     

Motility inOscillatoria salina as affected by different factors

All 3-10-d-old Oscillatoria salina filaments glide with the speed of 323-330 microm/min (BG 11 medium, pH 7.5, 21 +/- 2 degrees C, continuous light intensity of approximately 30 micromol m(-2) s(-1)) in a culture chamber. However, a time bound progressive decrease in gliding speed and in percentage of gliding filaments occurred, depending upon the severity of different stress factors studied, viz. water stress (2-8% agarized media, liquid media with 0.2-1 mol/L NaCl, blot-dryness of filaments for > or = 5 min), temperature shock (5, 40 degrees C for > or = 5 min; 35 degrees C for > or = 15 min), darkness and low light intensity (2, 10 micromol m(-2) s(-1)), UV exposure (0.96-3.84 kJ/m2), pH extremes (< or = 6.5 and > or = 9.5), lack of all nutrients from liquid medium (double distilled water), presence of 'heavy' metals (1, 25 ppm Fe, Cu, Zn, Ni, Co, Hg) or organic substances in liquid medium (25, 250 ppm 2,4-D, captan, urea, DDT, thiourea). This feature of the alga (i.e. reduction in speed and percentage of gliding filaments depending upon severity of stress conditions) may thus be suggested to be used in assessing water quality.     

Mucilage secretion and the movements of blue-green algae

Summary Recent discoveries of ultrastructures which might be involved in the gliding movements of blue-green algae have been reviewed, and in the light of these discoveries the role of mucilage secretion in movement has been reconsidered. The formation and behaviour of mucilage rings in filaments ofAnabaena cylindrica is described. The behaviour of the mucilage rings indicates that each cell has an autonomous gliding mechanism which is capable of immediate reversal, and that the gliding mechanism is probably located over the whole surface, rather than at the ends, of the cells. It follows that if mucilage secretion is the cause of movement it must take place over the whole surface of the cell: but if the ends of the cell are the sites of mucilage secretion, as seems likely, then gliding movement must be performed by some other process.A rather remarkable clumping phenomenon is described which takes place in dense suspensions ofAnabaena. It results from the gliding movements of randomly orientated filaments made mutually adhesive by the mucilage which surrounds them.     

The influence of daylength,light intensity and temperature on the growth rates of planktonic blue-green algae

The in vitro growth rates under continuous light of the four dominant blue-green algae in Lough Neagh, Anabaena flos-aquae Bréb., Aphanizomenon flos-aquae Ralfs fa. gracile Lemm., Oscillatoria agardhii Gom. and Oscillatoria redekei van Goor were slower than in situ rates from Lough Neagh that had been corrected for hours of light received by the algae. However, by culturing on a 6: 18 light-dark cycle in vitro growth rates were obtained that were similar to the in situ rates. Under continuous light small species showed the fastest growth with Oscillatoria redekei the dominant species. However, this pattern was almost completely reversed under the light-dark cycle with Oscillatoria redekei only exhibiting the fastest growth rate under low light conditions. This observation showed agreement with Lough Neagh field data which showed that Oscillatoria redekei reached its maximum crop in April while the other three species were dominant during the summer months. Compared to the generally assumed high thermal tendency of blue-green algae the temperature maxima of the four species were low. No growth was observed at 35°C for any species while Anabaena flos-aquae was severely inhibited at 25°C.     

Diel Migrations of Microorganisms within a Benthic, Hypersaline Mat Community

We studied the diel migrations of several species of microorganisms in a hypersaline, layered microbial mat. The migrations were quantified by repeated coring of the mat with glass capillary tubes. The resulting minicores were microscopically analyzed by using bright-field and epifluorescence (visible and infrared) microscopy to determine depths of coherent layers and were later dissected to determine direct microscopic counts of microorganisms. Microelectrode measurements of oxygen concentration, fiber optic microprobe measurements of light penetration within the mat, and incident irradiance measurements accompanied the minicore sampling. In addition, pigment content, photosynthesis and irradiance responses, the capacity for anoxygenic photosynthesis, and gliding speeds were determined for the migrating cyanobacteria. Heavily pigmented Oscillatoria sp. and Spirulina cf. subsalsa migrated downward into the mat during the early morning and remained deep until dusk, when upward migration occurred. The mean depth of the migration (not more than 0.4 to 0.5 mm) was directly correlated with the incident irradiance over the mat surface. We estimated that light intensity at the upper boundary of the migrating cyanobacteria was attenuated to such an extent that photoinhibition was effectively avoided but that intensities which saturated photosynthesis were maintained through most of the daylight hours. Light was a cue of paramount importance in triggering and modulating the migration of the cyanobacteria, even though the migrating phenomenon could not be explained solely in terms of a light response. We failed to detect diel migration patterns for other cyanobacterial species and filamentous anoxyphotobacteria. The sulfide-oxidizing bacterium Beggiatoa sp. migrated as a band that followed low oxygen concentrations within the mat during daylight hours. During the nighttime, part of this population migrated toward the mat surface, but a significant proportion remained deep.