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G protein and diacylglycerol regulate metamorphosis of planktonic molluscan larvae.

The regulatory guanine nucleotide binding protein (G protein) activators cholera toxin and the GTP analog 5-guanylyl imidophosphate, the second messenger diacylglycerol, and certain diamino acids all facilitate (amplify) the settlement and metamorphic responses of planktonic larvae of Haliotis rufescens (marine mollusc) to morphogenetic chemical stimuli. In contrast, the G protein-inhibiting analog guanosine 5'-O-[beta-thio]diphosphate inhibits facilitation by L-alpha,beta-diaminopropionic acid but does not block facilitation by diacylglycerol. Diacylglycerol, cholera toxin, and the guanine nucleotide analogs alone neither induce the settlement and metamorphosis of the larvae nor do they inhibit induction of metamorphosis by gamma-aminobutyric acid. These results thus establish the existence of separate regulatory and inductive pathways controlling larval metamorphosis in response to two classes of exogenous chemical signals from the environment. The regulatory pathway, operating independently through a G protein-diacylglycerol cascade apparently controlled by facilitating diamino acids in the water column, can amplify the larval responsiveness to inducers of metamorphosis. This mechanism may have adaptive significance in the recognition and selection of favorable habitats for metamorphosis of the larvae. Similar regulatory pathways, based on exogenous control of a G protein-diacylglycerol cascade, may govern responsiveness to stimuli in other sensory and developmental systems.

Journal Article↗

A new rubisco-like protein coexists with a photosynthetic rubisco in the planktonic cyanobacteria Microcystis.

Two genes encoding proteins related to large subunits of Rubisco were identified in the genome of the planktonic cyanobacterium Microcystis aeruginosa PCC 7806 that forms water blooms worldwide. The rbcL(I) gene belongs to the form I subfamily typically encountered in cyanobacteria, green algae, and land plants. The second and newly discovered gene is of the form IV subfamily and widespread in the Microcystis genus. In M. aeruginosa PCC 7806 cells, the expression of both rbcL(I) and rbcL(IV) is sulfur-dependent. The purified recombinant RbcL(IV) overexpressed in Escherichia coli cells did not display CO(2) fixation activity but catalyzed enolization of 2,3-diketo-5-methylthiopentyl-1-phosphate, and the rbcL(IV) gene rescued a Bacillus subtilis MtnW-deficient mutant. Therefore, the Microcystis RbcL(IV) protein functions both in vitro and in vivo and might be involved in a methionine salvage pathway. Despite variations in the amino acid sequences, RbcL(IV) shares structural similarities with all members of the Rubisco superfamily. Invariant amino acids within the catalytic site may thus represent the minimal set for enolization, whereas variations, especially located in loop 6, may account for the limitation of the catalytic reaction to enolization. Even at low protein concentrations in vitro, the recombinant RbcL(IV) assembles spontaneously into dimers, the minimal unit required for Rubisco forms I-III activity. The discovery of the coexistence of RbcL(I) and RbcL(IV) in cyanobacteria, the ancestors of chloroplasts, enlightens episodes of the chaotic evolutionary history of the Rubiscos, a protein family of major importance for life on Earth.

Amino Acid Sequence↗

Laser impact on marine planktonic diatoms: an experimental study using a flow cytometry system.

A flow cytometry system was used to evaluate the impact of pulsed laser irradiations from an Nd:YAG laser on two marine coastal water diatoms, Chaetoceros gracilis and Skeletonema costatum. Three flow speeds, i.e. 9, 18 and 27 ml min-1 and three laser fluences, i.e. 0.025, 0.05 and 0.1 J cm-2 pulse-1 were tested during this study. The reduction in cell density and chlorophyll a (chl a) concentrations were monitored by reference to non-irradiated samples as controls. Upon irradiation, the cell density and the chl a concentrations became reduced significantly compared to the control (one way ANOVA p < 0.001 for the cell density in both the species and p < 0.05 for chl a concentrations in both species). A maximum mortality of 0.77 log10 (about 83%) for C. gracilis and 0.68 log10 (about 78%) for S. costatum was observed at 9 ml min-1 flow speed and 0.1 J cm-2 laser fluence. The maximum reduction observed in the chl a concentration was about 26% (control 0.413 and sample 0.306 mg ml-1) for C. gracilis and 27% (control 0.222 and sample 0.16 mg ml-1) for S. costatum, when the flow rate was 9 ml min-1 and the fluence 0.1 J cm-2. In general, mortality increased with an increase in the laser fluence. The results thus show if the cooling water is laser-irradiated to mitigate biofouling, this could result in significant damage to the planktonic flora of the flowing seawater system, which in turn might reduce algal biofilm formation on industrially important structures. The reduction in the chl a concentration showed that the laser irradiations also could result in a significant reduction in the primary productivity of the cooling water.

Analysis of Variance↗

Chlorophyll Turnover in Skeletonema costatum, a Marine Plankton Diatom.

[(3)H]- and delta-[(14)C]Aminolevulinic acids were incorporated into the chlorophylls of Skeletonema costatum, a marine plankton diatom. In the stationary phase of growth, the tetrapyrrole-based pigments reached steady-state labeling after 10 hours. Under conditions of exponential cell division and chlorophyll accumulation, (3)H was rapidly lost from the labeled chlorophylls and was replaced with (14)C derived from delta-[4-(14)C]aminolevulinic acid. The kinetics of isotope dilution suggests recycling of tetrapyrrole precursors and/or two pigment pools, containing both chlorophyll a and chlorophyllide c, one which turns over rapidly (10 hours) and another which turns over more slowly (100 hours). Calculation of turnover times varied from 3 to 10 hours for chlorophyll a and from 7 to 26 hours for chlorophyllide c. The data suggest the dynamics of chlorophyll metabolism in S. costatum and explain the diatom's ability to undergo light-shade adaptation within a generation time.

Journal Article↗

Interactions between planktonic microalgae and protozoan grazers.

For an algal bloom to develop, the growth rate of the bloom-forming species must exceed the sum of all loss processes. Among these loss processes, grazing is generally believed to be one of the more important factors. Based on numerous field studies, it is now recognized that microzooplankton are dominant consumers of phytoplankton in both open ocean and coastal waters. Heterotrophic protists, a major component of microzooplankton communities, constitute a vast complex of diverse feeding strategies and behavior which allow them access to even the larger phytoplankton species. A number of laboratory studies have shown the capability of different protistan species to feed and grow on bloom-forming algal species. Because of short generation times, their ability for fast reaction to short-term variation in food conditions enables phagotrophic protists to fulfill the function of a heterotrophic buffer, which might balance the flow of matter in case of phytoplankton blooms. The importance of grazing as a control of microalgae becomes most apparent by its failure; if community grazing controls initial stages of bloom development, there simply is no bloom. However, if a certain algal species is difficult to graze, e.g. due to specific defense mechanisms, reduced grazing pressure will certainly favor bloom development. The present contribution will provide a general overview on the interactions between planktonic microalgae and protozoan grazers with special emphasis on species-specific interactions and algal defense strategies against protozoan grazers.

Animals↗

Urotricha psenneri n. sp. and Amphileptus piger (Vuxanovici, 1962) n. comb., two planktonic ciliates (Protozoa, Ciliophora) from an oligotrophic lake in Austria.

Two euplanktonic ciliates, Urotricha psenneri n. sp. (Prostomatida) and Amphileptus piger (Vuxanovici, 1962) n. comb. (Pleurostomatida), were discovered in the surface plankton of the oligotrophic Lake Traunsee in Austria. Their morphology and infraciliature were studied in live cells as well as in specimens impregnated with protargol and silver nitrate. Urotricha psenneri is a small urotrichid, less than 50 microm length and with a single caudal cilium. It is unique in having (i) a massive oral basket projecting as a conspicuous bulge with cylindrical microfibrillar annulus and (ii) a curved brosse row 1 in the broad, barren circumoral area. Amphileptus piger (Vuxanovici, 1962) is about 55 x 13 microm in vivo, has two macronuclear nodules with a single micronucleus in between in the posterior body half, has a single contractile vacuole with a terminal excretory pore, and few, but thick and thus highly conspicuous extrusomes. The amphileptid ciliary pattern (spica) is difficult to recognise due to the widely spaced basal bodies.

Animals↗

Comparison of pure cultures and natural assemblages of planktonic photosynthetic sulfur bacteria by low molecular mass RNA fingerprinting.

Pure cultures of phototrophic sulfur bacteria were compared to natural populations that bloom in karstic lakes by electrophoretic analysis of low molecular mass RNA molecules (lmwRNA) and microscopy. Similarities between dominant community members, field isolates and reference strains were established by comparing the lmwRNA band patterns through dendrograms produced with Euclidean distances and the average linkage clustering method, by a single, quick, one-step method. The fingerprinting analysis had three objectives: (i) to compare microbial assemblages from different geographical locations, (ii) to compare those organisms which grow in pure culture to those forming planktonic blooms and (iii) to give a preliminary view of the identity of the predominant community members. The lmwRNA analysis yielded a number of clusters consistent with the microscopic observations and allowed rapid comparison of the microbial communities without the need to isolate their individual components. The fingerprints of natural communities were different from most of the laboratory strains tested. Purple sulfur bacteria responsible for the blooms analyzed in karstic lakes were more closely related to the Thiocystis group than to the classical strains extensively studied in the laboratory.

Journal Article↗

Planktonic foraminifera of the California Current reflect 20th-century warming.

It is currently unclear whether observed pelagic ecosystem responses to ocean warming, such as a mid-1970s change in the eastern North Pacific, depart from typical ocean variability. We report variations in planktonic foraminifera from varved sediments off southern California spanning the past 1400 years. Increasing abundances of tropical/subtropical species throughout the 20th century reflect a warming trend superimposed on decadal-scale fluctuations. Decreasing abundances of temperate/subpolar species in the late 20th century indicate a deep, penetrative warming not observed in previous centuries. These results imply that 20th-century warming, apparently anthropogenic, has already affected lower trophic levels of the California Current.

Animals↗

Fish vision and the detection of planktonic prey.

Planktivorous sunfish of various sizes were studied to ascertain whether growth-related changes in the retina are related to the ability to capture small planktonic crustaceans. Behaviorally, the larger fish detected and captured crustaceans that subtended smaller visual angles. Histological examination of the retinas revealed that the distance between cones, measured in minutes of visual angle, decreased as the animals grew, suggesting that the larger retinas could resolve smaller objects. These correlated behavioral and anatomical results suggest that improved visual resolution contributes to improved predation. This finding provides a selective advantage for the continuous retinal growth noted in many fish.

Animals↗

Patchiness in the distribution of planktonic heterotrophic bacteria in lakes.

By using conventional dilution plating procedures, the heterotrophic planktonic bacteria in Lake Washington were found to be distributed in small-scale (<1.0 ml) and large-scale (>1 m in horizontal distance) patches. Recommendations are made on sampling and enumeration procedures to minimize the effect of patchiness on obtaining an accurate estimate of bacterial numbers.

Journal Article↗

Microcomputer-assisted biomass determination of plankton bacteria on scanning electron micrographs.

Although biovolume is a better measure of biomass than is cell number, biovolumes have rarely been measured because their evaluation is extremely time-consuming. We developed a microcomputer system that assists cell size measurements on images of filtered plankton: scanning electron micrograph negatives were projected on a digitizer field, bacterial length and width were marked by a cursor, and coordinates were directly transferred to an MOS 6502 microcomputer (KIM 1). The dialogue program BABI organized and controlled the digitizer measurements in cooperation with the user, enabled corrections, and printed out results with 95% confidence limits and sample description. The time for scanning electron micrograph preparation was reduced to 15 min (quick transfer to Freon 113 during filtration and air drying). Altogether, this biovolume determination took about 2.5 h for confidence limits of +/-15%. Examples are given for applications of the method: (i) comparison of 10 lakes (with specific activities for glucose uptake and for heterotrophic CO(2) fixation); (ii) ranges of biomass parameters in one lake; (iii) diurnal cycles (with synchronizing effects, uptake of algal exudates, and calculation of daily growth). This method is discussed in relation to other biomass methods (epifluorescent microscopy, lipopolysaccharide technique, frequency of dividing cells) and the problem of biovolume-to-carbon conversions.

Journal Article↗

Sensitivity of an oligotrophic lake planktonic bacterial community to oxygen stress.

Dissolved oxygen at approximately four times normal saturation (42 mg liter) inhibited the growth and metabolism of summer planktonic bacteria in the surface water of alpine oligotrophic Mountain Lake (Giles County, Va.). Data were derived from growth of CFU on membrane filters, d-[U-C]glucose incorporation into the extractable lipid of these CFU, and respiration and assimilation of d-[U-C]glucose by lake water samples. Statistically significant (alpha < 0.05) differences were not detected in either CFU or C incorporation in lipid when superoxide dismutase (30 U ml) or catalase (130 U ml) was added to the medium. Thus, exogenous oxygen by-products apparently are not responsible for the observed inhibition of growth and metabolism.

Journal Article↗

Influence of cyanobacterial hyperscum on heterotrophic activity of planktonic bacteria in a hypertrophic lake.

The response of the planktonic heterotrophic bacterial community to the buildup and breakdown of a semipermanent, crusted, floating cyanobacterial mat, or hyperscum, that covered 1 to 2 ha was studied in a hypertrophic lake (Hartbeespoort Dam, South Africa). The initial response of bacteria in the main basin to the release of dissolved organic carbon (DOC) from the hyperscum 1 km away was an increase in activity per cell from 35 x 10 to 153 x 10 mug of C cell h for total cell counts, while activity per cell for metabolically active cells increased from 19 x 10 to 85 x 10 mug of C cell h. No major population growth occurred at this stage. Later, with the continuous supply of DOC from the hyperscum, total bacterial numbers increased from 6.6 x 10 to 20 x 10 cells ml, while the activity per cell declined. Metabolically active bacteria followed the same trend. Shorter-term DOC increases caused only increases in bacterial activity per cell. The data from Hartbeespoort Dam demonstrate an interesting and little-documented mechanism by which aquatic bacteria respond to increased DOC concentration and which may be universal for aquatic systems.

Journal Article↗

Temperature characteristics of photosynthetic and heterotrophic activities: seasonal variations in temperate microbial plankton.

The seasonal variation in temperature characteristics of photosynthetic and heterotrophic activities in the microbial plankton of Bedford Basin, Nova Scotia, was investigated. Measurements were made of the photosynthetic uptake of [C]bicarbonate and its incorporation into cellular protein as well as the heterotrophic uptake of H-labeled amino acids and their incorporation into cellular protein. Activity-temperature curves were analyzed objectively by nonlinear estimation of parameters from various mathematical models. Over the seasonal cycle, the cardinal temperatures and a parameter formally equivalent to the thermodynamic enthalpy of activation for most of the four processes measured were positively correlated with the water temperature. The temperature sensitivity of metabolic activity (i.e., change in activity per unit change in temperature) was indexed by the tangent to the activity-temperature curves. When this index was expressed in dimensionless form by normalization to the scaling factor of the activity-temperature curves, the resulting relative temperature sensitivity, evaluated at the prevailing temperature, proved to be statistically invariant throughout the year. During the height of the spring bloom, the water temperature (-0.3 degrees C) was not so low as to inhibit metabolic activity of either the phytoplankton or the bacterioplankton. The evidence suggests that heterotrophic utilization of products is not suppressed during the spring phytoplankton bloom.

Journal Article↗

Production rate of planktonic bacteria in the north basin of lake biwa, Japan.

Vertical and seasonal variations in the cell number and production rate of planktonic bacteria were investigated at a pelagic site (water depth, ca. 72 m) of the north basin of Lake Biwa during April to October 1986. The [methyl-H]thymidine uptake rate into a cold trichloroacetic acid-insoluble fraction and the frequency of dividing cells (FDCs) were measured for each sample as indices of the bacterial production rate. The seasonal data of bacterial number, thymidine uptake rate, and bacterial growth rate based on the FDCs were correlated with one another (rank correlation analysis, P < 0.05). These bacterial variables were not correlated positively with the chlorophyll a concentration. Vertically, the maxima of both bacterial number and the thymidine uptake rate were found in the euphotic zone. The direct counting of bacteria and the measurements of thymidine uptake rate combined with the size-fractionation method revealed that more than 90% of the bacterial biomass and production rate were attributed to unattached bacteria throughout the investigation period. The carbon flux estimates of bacterial production were less certain due to the variability of the conversion factor for the thymidine uptake method and that of the calibration for the FDC method, but even when the conservative range of bacterial net production rate was used (5 to 60 mug of carbon per liter per day), it can be suggested that bacterial net production in the investigated area was a significant fraction (ca. 30%) of the level of the primary production rate in the same water basin.

Journal Article↗

Effects of fixation on cell volume of marine planktonic protozoa.

The effects of fixation on the cell volume of marine heterotrophic nanoflagellates and planktonic ciliates were investigated. Decreases in cell volume depended on the combination of the protozoan taxa and the particular fixative. For a particular fixative and protozoan species, degree of shrinkage was independent of physiological state. The volume of fixed cells was found to be approximately 20 to 55% lower than the cell volume of live organisms. For the heterotrophic microflagellates, the fixatives ranked, in order of decreasing effect on cell volume, as glutaraldehyde, formaldehyde, acid Lugol's solution, and modified van der Veer solution. With oligotrichous ciliates and a tintinnid ciliate, formaldehyde caused less shrinkage than glutaraldehyde or acid Lugol's solution. With the aldehyde fixatives, the microflagellates were found to shrink more than the ciliates. Differential effects of fixation on cell volumes may result in an underestimation of the biomass of certain protozoan taxa in natural samples.

Journal Article↗

Identification of Vibrio splendidus as a Member of the Planktonic Luminous Bacteria from the Persian Gulf and Kuwait Region with luxA Probes.

Hybridization probes specific for the luxA genes of four groups of luminous bacteria were used to screen luminous isolates obtained from the Persian Gulf, near Al Khiran, Kuwait Nine of these isolates were identified as Vibrio harveyi, a commonly encountered planktonic isolate, while three others showed no hybridization to any of the four probes (V. harveyi, Vibrio fischeri, Photobacterium phosphoreum, or Photobacterium leiognathi) under high-stringency conditions. Polymerase chain reaction amplification was used to prepare a luxA probe against one of these isolates, K-1, and this probe was screened under high-stringency conditions against a collection of DNAs from luminous bacteria; it was found to hybridize specifically to the DNA of the species Vibrio splendidus. A probe prepared against the type strain of V. splendidus (ATCC 33369) was tested against the collection of luminous bacterial DNA preparations and against the Kuwait isolates and was found to hybridize only against the type strain and the three unidentified Kuwait isolates. Extensive taxonomic analysis by standard methods confirmed the identification of the 13 isolates.

Journal Article↗

Iron-Stimulated N(2) Fixation and Growth in Natural and Cultured Populations of the Planktonic Marine Cyanobacteria Trichodesmium spp.

In light of recent proposals that iron (Fe) availability may play an important role in controlling oceanic primary production and nutrient flux, its regulatory impact on N(2) fixation and production dynamics was investigated in the widespread and biogeochemically important diazotrophic, planktonic cyanobacteria Trichodesmium spp. Fe additions, as FeCl(3) and EDTA-chelated FeCl(3), enhanced N(2) fixation (nitrogenase activity), photosynthesis (CO(2) fixation), and growth (chlorophyll a production) in both naturally occurring and cultured (on unenriched oligotrophic seawater) Trichodesmium populations. Maximum enhancement of these processes occurred under FeEDTA-amended conditions. On occasions, EDTA alone led to enhancement. No evidence for previously proposed molybdenum or phosphorus limitation was found. Our findings geographically extend support for Fe limitation of N(2) fixation and primary production to tropical and subtropical oligotrophic ocean waters often characterized by Trichodesmium blooms.

Journal Article↗