Observations on the valve structure of marine species of the diatom genus Cocconeis Ehr.

The valve structure of three marine diatom species, Cocconeis molesta var. crucifera, C. dirupta and C. pellucida was studied by transmission and scanning electron microscopy. In all Cocconeis examined, the areolae are occluded by hymenes located near the internal openings in the raphid valve and near the internal openings in the araphid valves.     

Morphology of the marine epiphytic diatom Cocconeis heteroidea (Bacillariophyceae)

The morphology and fine valve structure of the marine epiphytic diatom Cocconeis heteroidea Hantzsch have been investigated. The entire frustule, including the internal and external structure of the raphid valve (RV) and araphid valve (AV), and the complete cingulum, are described using light microscopy and scanning and transmission electron microscopy, using a bleaching method. The strongly sigmoid raphe terminates in elongate hooked helictoglossae internally. The hymenes, with perforations arranged in a centric array, are located near the internal openings of the areolae in the RV. The striae in the AV consist of alveoli occluded by hymenes, that have perforations arranged in a parallel array and are located near the outer surface. The complete cingulum of AV consists of three open bands without fimbriae: a valvocopula, a copula with a ligula and a pleura with a small ligula. The RV has only a valvocopula which is open type and not fimbriate.     

Change in striation density and systematics ofCocconeis scutellum var.Ornata (Bacillariophyceae)

Cocconeis scutellum var.ornata Grun. from three localities of Japan was studied. The striation density in 10 μm showed a marked tendency to increase with the decrease of the valve length in both raphe and rapheless valves, and this tendency did not vary with locality or environmental condition. The striation densities of rapheless valves were 4–6 in 10 μm for a valve length of 40μm, 4–6.5 for 30 μm, 6–9 for 20μm and 6.5–11 for 15μm. Those of raphe valves were 10–11 in 10μm for a valve length of 40μm, 10–12 for 30μm, 11–14.5 for 20μm and 12.5–17 for 15μm. According to the range of changing value in striation density obtained by the present study,C. scutellum var.schmidti Frenguelli andC. japonica Schmidt are identical withC. scutellum var.ornata. Dedicated to Prof. Munenao Kurogi on the occasion of his academic retirement. Culture experiment in the present study was undertaken at the Institute of Algological Research, Faculty of Science, Hokkaido University at Muroran.     

Influence of diet on sex differentiation of Hippolyte inermis Leach (Decapoda: Natantia) in the field

The gut contents of the shrimp Hippolyte inermis were investigated for 1 year along a depth transect through a seagrass bed. Besides size, sex and weight of all individuals were recorded. The diets of immature and adult individuals were compared to detect any influence of food on sex development, since previous investigations indicated a correlation of the life cycle of this protandric species with the abundance of algal food in the environment, and laboratory experiments demonstrated the effect of diatoms of the genus Cocconeis on the direct development of females. Results indicated that the shrimp is an opportunistic herbivore, able to feed on both plant and animal items, with a preference for macroalgae and diatoms present on the leaves of Posidonia oceanica. Small females, deriving from direct differentiation, had a diet significantly different from that of males. The difference was due to a larger abundance of microalgae in the guts of young females. The influence of microalgal food on the sex reversal mechanism of this species, previously detected through laboratory experiments, was demonstrated to control the life cycle of H. inermis in the field.     

Metabolite profiling and assessment of metabolome compartmentation of soybean leaves using non-aqueous fractionation and GC-MS analysis

In the present study, non-aqueous fractionation (NAQF) and GC-MS were used to obtain a spatially resolved view of metabolism in mature leaves of soybean (Glycine max Merr.). NAQF of lyophilized soybean leaves was performed using CCl₄-n-heptane and ultracentrifugation that yielded a gradient comprised of six fractions. Chlorophyll content, and marker enzyme activities, phosphoenolpyruvate carboxylase (PEPC) and α-mannosidase, were utilized as stroma, cytosol and vacuole markers, respectively. GC-MS analyses of each fraction resulted in the identification of around 100 different metabolites. The distribution of these identified compounds showed a decreasing order from the vacuole to cytosol to chloroplast stroma. In other words, a greater number of identified compounds were found in the vacuole when compared to the cytosol or stroma. Levels of sugars, organic acids and fatty acids showed greater relative abundances in the vacuole with 50, 55, and 50% of the respective pools. A greater relative abundance of amino acids was observed in the cytosol where 45% of the total of amino acids content was recorded. The relatively large pool of sugars and phenolic acids in the vacuole compartment implies high levels of starch metabolism and phenylpropanoid biosynthesis. The low amino acids pool, on the other hand, suggests low nitrogen accumulation in the leaves of soybean. Hierarchical cluster analysis on the most abundant metabolites revealed three clusters containing 10, 20, and 2 of the 32 selected metabolites. The data were discussed in term of NAQF and GC-MS analysis of soybean mature leaves, and also in term of distribution and compartmentation of metabolites at subcellular levels.     

Metabolite profiling of saponins in <Emphasis Type="Italic">Balanites</Emphasis><Emphasis Type="Italic">aegyptiaca</Emphasis> plant tissues using LC (RI)-ESI/MS and MALDI-TOF/MS

Saponins are a major family of secondary metabolites which consist of a sugar moiety glycosidically linked to a hydrophobic aglycone (sapogenin). In recent years the interest in saponins has increased significantly because of their diverse properties as natural detergents and foaming agents, their cardiac, immunostimulating, and anti-cancer activity, as well as other health promoting functions. This study deals with metabolitic analysis of saponins from methanolic extracts of fruit mesocarp (ME), seed kernel (KE) and root (RE) of Balanites aegyptiaca (L.) Del. (desert date) plant grown in Israel using LC (RI)-ESI/MS and MALDI-TOF/MS. The structural assignment was carried out by fragmentation experiments of LC (RI)-ESI/MS and literature data. The study has revealed that, all together, twenty-four furostanol saponins were found in ME, KE and RE. Of these, four saponins are found only in ME, five only in KE and six only in RE. Diosgenin was found to be the sole aglycone in all the saponins. The smallest saponin (MW 740 Da) was found with two sugar units (glucose) and the largest saponin (MW 1678 Da) was found with eight sugar units (5 glucose, 2 rhamnose and 1 xylose) attached to diosgenin. The results suggest that MALDI-TOF/MS with positive ion mode is particularly effective for determining the metabolites of saponins in B. aegyptiaca plant tissues. MALDI-TOF/MS not only verified the results of the LC (RI)-ESI/MS, but also identified additional saponins that are now systematically organized in a database of B. aegyptiaca saponins. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Effect of ageing on survival of benthic diatom propagules

A comparison of viable benthic diatom propagules based on the observations recorded immediately and after 5 years of ageing at 5 °C is presented. The number of viable benthic diatom propagules decreased with ageing. However, they exhibited an apparently longer lag phase. Although diatoms belonging to the genera Amphora, Navicula and Thalassiosira were dominant during immediate observation, only Amphora and Navicula survived the ageing process. The non-viability of Thalassiosira indicates that ageing for five years was beyond its critical period of survival. The other diatom genera that survived the ageing process were Odontella and Grammatophora.     

Inorganic-carbon uptake by the marine diatom Phaeodactylum tricornutum

Air-grown cells of the marine diatom Phaeodactylum tricornutum showed only 10% of the carbonic-anhydrase activity of air-grown Chlamydomonas reinhardtii. Measurement of carbonic-anhydrase activity using intact cells and cell extracts showed all activity was intracellular in Phaeodactylum. Photosynthetic oxygen evolution at constant inorganic-carbon concentration but varying pH showed that exogenous CO₂ was poorly utilized by the cells. Sodium ions increased the affinity of Phaeodactylum for HCO ₃ ^- and even at high HCO ₃ ^- concentrations sodium ions enhanced HCO ₃ ^- utilization. The internal inorganic-carbon pool (HCO ₃ ^- +CO₂] was measured using a silicone-oil-layer centrifugal filtering technique. The internal [HCO ₃ ^- +CO₂] concentration never exceeded 15% of the external [HCO ₃ ^- +CO₂] concentration even at the lowest external concentrations tested. It is concluded that an internal accumulation of inorganic carbon relative to the external medium does not occur in P. tricornutum.Abbreviation Hepes 4-(2-hydroxyethyl)-1-piperazineethane-sulfonic acid     

Analysis of the peptide carrier in the scutellum of barley embryos by photoaffinity labelling

The preparation of a phenylalanine analogue containing an azido group and its incorporation into dipeptides is described. Peptides modified in this way are taken up into barley (Hordeum vulgare L.) scutella via the previously characterized peptide-transport system. Photoactivation of modified peptides in the presence of isolated scutella resulted in irreversible inhibition of peptide uptake in a concentration-dependent manner. Transport of other solutes which share a common mechanism of energy coupling, but which are transported via distinct carriers, was not inhibited after photo-derivatization of scutella with the modified peptides. Derivatization of isolated scutellar tissue with a ¹⁴C-labelled peptide analogue, resulted in incorporation of label into two proteins of M_r = 54000 and 41000. Scutellar tissue from early-germinating seeds, which do not show active peptide uptake, did not incorporate label into these polypeptides. It is concluded that these proteins are components of the barley peptide-transport system.Abbreviations Ala alanine - Gly glycine - PAGE polyacrylamide gel electrophoresis - Phe phenylalanine - Pro proline - SDS sodium dodecyl sulphate This work was supported by a grant from the Agricultural and Food Research Council.     

Analysis of variants affecting the catalase developmental program in maize scutellum

Summary The catalase of maize scutella is coded for by two loci, Cat1 and Cat2, which are differentially expressed in this tissue during early seedling growth. Two variant lines have been previously identified in which the developmental program for the expression of the Cat2 structural gene in the scutellum has been altered. Line R6–67 exhibits higher than normal levels of CAT-2 catalase in this tissue after four days of postgerminative growth. This phenotype is controlled by a temporal regulatory gene designated Car1. Line A16 exhibits a CAT-2 null phenotype. Further analysis of Car1 verifies the initial indication that it is trans-acting and exhibits strict tissue (scutellum) specificity. A screen of other available inbred lines uncovered eight additional catalase high-activity lines. All eight lines exhibit significantly higher than normal levels of CAT-2 protein. Two of these lines have been shown to be regulated by Car1 as in R6–67. Another line (A338) uncovered during the screen exhibits a null phenotype for CAT-2 protein and resembles A16. Catalase activity levels are low in the scutellum and no CAT-2 CRM (cross-reacting material) is present in the tissues of this line. Also, unlike most maize lines, CAT-2 cannot be induced in the leaf tissue of A338 upon exposure to light. Finally, a single line (A337), demonstrating a novel catalase developmental program, was identified.     

Nitrogen fixation by the benthic freshwater cyanobacterium Lyngbya wollei

The ability of the benthic cyanobacterium Lyngbya wollei to fix nitrogen was studied using field samples and axenic cultures. L. wollei was collected and isolated from Lake Okeechobee, Florida, where it forms extensive mats. Rates of acetylene reduction up to 39.1 nmol mg dry wt⁻¹ h⁻¹ were observed for field samples. The maximum observed rate of acetylene reduction in axenic laboratory cultures was 200 nmol mg dry wt⁻¹ h⁻¹. Aerobic conditions limited nitrogen fixation activity, but dark/light cycles promoted the development of activity. Reduced oxygen levels appeared to be required for the development of significant levels of nitrogenase activity. The level of irradiance also had a significant impact on the level of activity. The potential significance of nitrogen fixation to Lyngbya production is discussed.     

The benthic invertebrates of the Salton Sea: distribution and seasonal dynamics

The Salton Sea, California's largest inland water body, is an athalassic saline lake with an invertebrate fauna dominated by marine species. The distribution and seasonal dynamics of the benthic macroinvertebrate populations of the Salton Sea were investigated during 1999 in the first survey of the benthos since 1956. Invertebrates were sampled from sediments at depths of 2–12 m, shallow water rocky substrates, and littoral barnacle shell substrates. The macroinvertebrates of the Salton Sea consist of a few invasive, euryhaline species, several of which thrive on different substrates. The principal infaunal organisms are the polychaetes Neanthes succinea Frey & Leuckart and Streblospio benedicti Webster, and the oligochaetes Thalassodrilides gurwitschi Cook, T. belli Hrabe, and an enchytraeid. All but Neanthes are new records for the Sea. Benthic crustacean species are the amphipods Gammarus mucronatus Say, Corophium louisianum Shoemaker, and the barnacle Balanus amphitrite Darwin. Neanthes succinea is the dominant infaunal species on the Sea bottom at depths of 2–12 m. Area-weighted estimates of N. succinea standing stock in September and November 1999 were two orders of magnitude lower than biomass estimated in the same months in 1956. During 1999, population density varied spatially and temporally. Abundance declined greatly in offshore sediments at depths >2 m during spring and summer due to decreasing oxygen levels at the sediment surface, eventually resulting in the absence of Neanthes from all offshore sites >2 m between July and November. In contrast, on shoreline rocky substrate, Neanthes persisted year round, and biomass density increased nearly one order of magnitude between January and November. The rocky shoreline had the highest numbers of invertebrates per unit area, exceeding those reported by other published sources for Neanthes, Gammarus mucronatus, Corophium louisianum, and Balanus amphitrite in marine coastal habitats. The rocky shoreline habitat is highly productive, and is an important refuge during periods of seasonal anoxia for Neanthes and for the other invertebrates that also serve as prey for fish and birds.     

Production potential of eicosapentaenoic acid by the diatom Nitzschia laevis

To investigate the production potential of eicosapentaenoic acid (EPA) by the diatom Nitzschia laevis, the growth characteristics and fatty acid composition of the cells were studied under photoautotrophic, mixotrophic and heterotrophic conditions of growth. The specific growth rate and maximum biomass concentration were respectively 0.466 d^–1 and 2.27 g l^–1 for mixotrophic culture, 0.344 d^–1 and 2.04 g l^–1 for heterotrophic culture, and 0.167 d^–1 and 0.5 g l^–1 for photoautotrophic culture, respectively. As for EPA production, the yield and productivity were respectively 52.32 mg l^–1 and 10.46 mg l^–1 d for mixotrophic culture, 35.08 mg l^–1 and 6.37 mg l^–1 d for heterotrophic culture, and 6.78 mg l^–1 and 3.39 mg l^–1 d for photoautotrophic culture, respectively. Results suggest that mixotrophic culture is the most suitable growth mode for the production of EPA by the diatom Nitzschia laevis. The results are useful for the development of a cost-effective fermentation process for EPA production by Nitzschia laevis.     

Effect of Lawsonia inermis on tumor expression induced by Dalton’s lymphoma ascites in Swiss albino mice

The purpose of this study was investigating experimentally the possible antitumor effect of ethanol extract of root of Lawsonia inermis against Dalton’s lymphoma ascites (DLA) bearing mice.Mice were administered with L. inermis at a dosage of 180 mg/kg of body weight for 15 days after 24 h of DLA inoculation. The ethanolic root extract of L. inermis reverted the increased number of the WBC count, platelets and lymphocytes and decreased the number of the RBC count, hemoglobin content and monocytes. The effect of root extract of L. inermis also increased the pathophysiological marker enzyme, lipid profile and decreased the enzymic and non enzymic antioxidants. A histopathological result shows the loss of liver hepatocytes and kidney architecture in DLA bearing mice. However, mice treatment with L. inermis extract improves the liver and kidney function and rearranges more or less normal architecture. The present work indicates that the ethanol extract of L. inermis exhibited significant antitumor activity.     

Convergence of a cryptic saddle pattern in benthic freshwater fishes

Synopsis Many North American stream fishes have a similar color pattern of four dark saddles against a light background. An interesting feature of the pattern, in addition to its widespread taxonomic distribution, is its consistent configuration. The interval between the first and second saddle is usually the largest, and the last (third) interval is the smallest. All saddled North American freshwater fishes live on uneven, rocky substrates, and nearly all live in flowing water. It is hypothesized that these fishes achieve crypsis through disruptive coloration; the light spaces between the saddles mimic rocks and the dark saddles appear as shadows or gaps between rocks. Saddles are spaced unevenly because rocks in streams are a mixture of sizes; a fish that mimics a series of rocks of similar sizes is more conspicuous than one that mimics rocks of different sizes. The placement of saddles was measured on five North American species. In four of five North American species measured (a sculpin and three darters), the longest spaces are towards the head where the body is also the widest, this is thought to enhance crypsis because pieces of gravel tend to be round or square. In the madtom, the saddle pattern tends more towards even spacing. The madtom may not rely on camouflage to the same extent as other species examined because of decreased predation pressure associated with being nocturnal and possessing sharp spines and venom glands.     

Acclimation of the diatom Stephanodiscus neoastraea and the cyanobacterium Planktothrix agardhii to simulated natural light fluctuations

Functional and structural characteristics of the photosynthetic apparatus were studied in the diatom Stephanodiscus neoastraea and the cyanobacterium Planktothrix agardhii which were grown semi-continuously under constant irradiance or under simulated natural light fluctuations. The light fluctuations consisted of 24 oscillations of exponentially increasing and decreasing irradiance over a 12-h light period. Maximum irradiance was 1100 μmol photons m⁻² s⁻¹ with the ratio of maximum to minimum intensities being 100, simulating Langmuir circulations with a ratio of euphotic to mixing depth of 1. S. neoastraea acclimated to the light fluctuations by doubling the number and halving the size of photosynthetic units (PS II) while the amount of chlorophylls and carotenoids remained unchanged. The chlorophyll-specific maximum photosynthetic rate was enhanced while the slope of the photosynthesis versus irradiance curves was not influenced by the light fluctuations. Acclimation of P. agardhii was mainly characterized by an increase in chlorophyll content. Both photosystems showed only little changes in number and size. Maximum photosynthetic rate, saturating irradiance and initial slope of the photosynthesis versus irradiance curves did not vary. Although both high and low light were contained in the fluctuating light, an analogy to low or high light acclimation was not found for the diatom nor for the cyanobacterium acclimated to light fluctuations. We suggest that the acclimation to fluctuating light is a response type outside the known scheme of low and high light acclimation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Uptake of free amino acids by the diatom,Melosira mediocris

Individual ¹⁴C-labelled amino acids are rapidly removed from dilute solution in artificial sea water (0.2 mol 1^–1) by suspensions of Meliosira medocris. The rate of disappearance of radioactivity corresponds closely to removal of primary amines as determined by measurement of the rate of decrease of fluorescamine-positive material. Net removal of naturally occurring free amino acids from the sea water habitat from which the alga was isolated is demonstrated using high performance liquid chromatography. Removal of amino acids from natural sources makes a significant contribution to the carbon requirements of the alga as well as supplying significant amounts of amino nitrogen.