ASCOPHYLLUM NODOSUM (PHAEOPHYTA) IN AXENIC CULTURE AND ITS RESPONSE TO THE ENDOPHYTIC FUNGUS MYCOSPHAERELLA ASCOPHYLLI AND EPIPHYTIC BACTERIA1

Ascophyllum nodosum (L.) Le Jol. was freed from bacteria and the endophytic fungus Mycosphaerella ascophylli Cotton by repeated treatment with chlorine solutions and grown in artificial seawater. Two types of axenic culture of different origin were obtained. Type 1 was developed from apices of A. nodosum collected in the sea. Type 2 was from plants which developed from adventitious embryos on rhizoids formed by type 1. This is the first time A. nodosum has been cultivated axenically. Growth of the axenic alga was increased by IAA, 21P and zeatin. Without external growth regulators some strains of the axenic alga deteriorated within a year; others developed a filamentous habit. Sulfur in a reduced state also stimulated growth. Addition of either glucose, mannose or mannitol to the medium caused the formation of calluslike layers of loosely packed colorless cells under the epidermis of the thalli and the epidermis was sloughed off. No increase in thallus length was noticed. Mycosphaerella ascophylli in axenic culture did not excude any substances stimulating growth of the alga, but that does not exclude an influence of the fungus on the alga in vivo. The fungus, when growing within the alga, seemed to have little influence on algal morphology. A bacterized but fungus-free A. nodosum was cultivated in an artificial seawater for 8 years. In the bacteria-free alga, the fungus protruded from the epidermis and evidently utilized the alga as a carbon source. The bacteria thus seem much more important than the fungus for normal growth of the Ascophyllum plant.     

Estimating growth and mortality rates from size data

Summary A method is presented for estimating rates of individual growth and population mortality utilizing average individual size at two times during a year. The model assumes a constant rate of mortality, Brody-Bertalanffy growth, a stationary age distribution, and recruitment confined to one month each year. A hypothetical example is presented to show the interrelationships of the growth and mortality constants, size at recruitment, asymptotic size and average individual size. Three examples are presented using data from the literature: Flathead sole (Hippoglossoides elassodon), a sea urchin(Echinus esculentus), and the crown-of-thorns starfish(Acanthaster planci). The method appears to be a means of obtaining reasonable approximations of growth and mortality rates for a variety of organisms.     

Marine antifoulants from bifurcaria bifurcata (phaeophyceae,cystoseiraceae) and other brown macroalgae

In this study, the antifouling activity of a series of extracts and linear diterpenes isolated from Bifurcaria bifurcata (Velley) Ross, a common brown alga of the Atlantic shores of Europe, and derivatives of these compounds was investigated. Antifouling assays with crude extracts from other brown algae, found abundantly along the coast of South Africa (Bifurcaria brassicaeformis, Bifurcariopsis capensis), the Atlantic shores of Europe (Halidrys siliquosa) and the coast of Mediterranean sea (Cladostephus verticillatus, Halopteris scoparia), are also reported. The fractions were tested in laboratory assays against representative species of the major groups of fouling organisms, viz. bacteria, fungi, diatoms, spores and zygotes of macroalgae and the blue mussel Mytilus edulis. Several components showed promising levels of activity. The high, albeit variable, level of antifouling activity suggests a potential for novel active ingredients in antifouling preparations.     

Proto-oncogene homologous sequences in the sea urchin genome

Using probes specific for several oncogenes/proto-oncogenes we have performed gel blot hybridization analyses of genomic DNA isolated from the sea urchinStrongylocentrotus droebachiensis. Probes prepared from v-erbB, v-myc, c-myb and v-fps were found to hybridize with discrete fragments of HindIII digested genomic DNA. In contrast, probes prepared from v-abl, v-fos, v-sis, v-src, and v-mos either hybridized with multiple fragments, indicating non-specific binding, or failed to hybridize at all above background levels. These results clearly demonstrate the presence of proto-oncogene homologous sequences in the sea urchin genome.     

TRANSFER OF PRODUCTS BETWEEN EPIPHYTIC MARINE ALGAE AND HOST PLANTS1,2

The red alga Smithora naiadum is normally found only as an epiphyle on the sea grasses Phyllospadix scouleri and Zostera marina. I used ³²P and ¹⁴CO₂ to examine the chemical communication between host and alga. Both ³²P and the product of ¹⁴CO₂ light fixation moved from the host to the alga. Reverse movement between host and epiphyte was also demonstrated. Part of this transfer occurred through the plant and part occurred by leakage from the host into the medium and subsequent uptake by the alga. Although plants were initially labeled in the light, transfer of ¹⁴C was light independent. Transfer of ¹⁴C-labeled products between host and epiphyte was also shown for Punctaria orbiculata and Z. marina; for Microdadia coulteri on Grateloupia doryphora, and between Gonimophyllum skottsbergii and Botryoglossum ruprechtiana. Epiphyte-host associations do not require a penetrating rhizoid for an exchange of the isotopes tested. By their proximity alone, epiphytic flora are apparently capable l exchanging products before these are diluted by the sea.     

Calcium-activated neutral proteinase in gametes of the sea urchinStrongylocentrotus droebachiensis

By Chromatographic separation of cytosol fraction of homogenates from gonads of females and males of green sea urchinStrongylocentrotus droebachiensis using an AcA-34 ultragel, protein fractions with mol. mass of 123 and 81 kDa are revealed, which have activity of calciumactivated proteinases. Dynamics of activity of the calcium-activated proteinases is studied during annual sexual cycles of the sea urchin. It is shown that the maximal activity of the studied enzyme in females and males of the sea urchin is present at the IV stage of gonad maturation, at the period of their trophic growth which is characterized by formation of stores of nutritient substances necessary for the subsequent development of embryos.     

Calcium-activated neutral proteinase in gametes of the sea urchinStrongylocentrotus droebachiensis

By Chromatographic separation of cytosol fraction of homogenates from gonads of females and males of green sea urchinStrongylocentrotus droebachiensis using an AcA-34 ultragel, protein fractions with mol. mass of 123 and 81 kDa are revealed, which have activity of calcium-activated proteinases. Dynamics of activity of the calcium-activated proteinases is studied during annual sexual cycles of the sea urchin. It is shown that the maximal activity of the studied enzyme in females and males of the sea urchin is present at the IV stage of gonad maturation, at the period of their trophic growth which is characterized by formation of stores of nutritient substances necessary for the subsequent development of embryos.     

Cover

ON THE COVER: Underwater photo showing luxurious growth of the obligately epizoic red alga Melanothamnus maniticola Woodworth, Frankovich & Freshwater sp. nov. on a West Indian Manatee (Trichechus manatus). This species has developmentally different rhizoids that are unique within Melanothamnus and may be an evolutionary adaptation for anchoring the thalli to manatee epidermis. Photo courtesy of Cathy Beck, USGS.     

Patterns of nocturnal movement of the long-spined sea urchin<Emphasis Type="Italic"> Diadema antillarum</Emphasis> (Philippi) in Gran Canaria (the Canary Islands,central East Atlantic Ocean)

We tagged individuals of the sea urchin Diadema antillarum (Philippi) around the island of Gran Canaria (The Canary Islands) during winter 2001–2002 using a new technique, consisting of the insertion of a hook fastened to a fishing line into the aboral pole (periproctal membrane). This allowed individual identification of tagged sea urchins. The goals were: (1) to quantify nocturnal movements and the homing behaviour of this echinoid on shallow rocky bottoms, and (2) to assess short term spatial and temporal variability of these movements. Tagged sea urchins displayed clear homing behaviour. The mean distance travelled at night was 3.7±1.2 m (range 1.0–5.1 m). Mean speed of nocturnal movement was 33±26 cm h^–1 (range 5–110 cm h^–1). We observed greater movement at midnight than at the beginning and the end of the night.Communicated by H.-D. Franke     

ATPase activity in plasmalemma-rich vesicles isolated by aqueous two-phase partitioning from Vicia faba mesophyll and epidermis: Characterization and influence of abscisic acid and fusicoccin

Plasmalemma-rich microsomal vesicles were prepared from whole leaf and acid-washed epidermal tissue of Vicia faba L. cv. Osnabrücker Markt by aqueous two-phase partitioning in dextran T-500 and polyethylenglycol 1350 aqueous phases. These vesicles were tightly sealed and predominantly right-side out, and contained a K⁺ -stimulated, mg²⁺-dependent and vanadate-sensitive ATPase. The enzyme from both tissues exhibited nearly identical properties: pH optimum 6.4, K_m for ATP 0.60 mM(whole leaf) and 0.67 mM (epidermis). V_max -480 nmol (mg protein)¹ min¹ (whole leaf) and 510 nmol (mg protein)¹ min¹ (epidermis), I₅₀ (Na₃,VO₄) 7.5 μM (whole leaf) and 15 μM (epidermis). The enzyme was not inhibited by NO₃(50 mM)or sodium azide (I mM). DCCD (20 μM) reduced enzyme activity to 50% (whole leaf) and 58% (epidermis), gramicidin S (20 μM) to 36% (whole leaf) and 41%(epidermis). Ca²⁺ inhibited the ATPase [I₅₀, C²⁺: 0.5 mM(whole leaf) and 0.8 mM(epidermis)]. Ca²⁺ inhibited the ATPase [I₅₀, C²⁺ 0.5 mM(whole leaf) und 0.8 (epidermis)]. The vanadate-sensitive ATPase from whole leaf and epidermal tissue was slightly but significantly stimulated by fusicoccin (FC) at a concentration (0.13 μM) promoting stomatal opening. The stimulation was not seen in the solubilized ATPase. Stomata of the cultivar used here were insensitive lo (±)ABA up to 2 μM level which is effective in most other cultivars and species. Likewise, at this concentration no effect of ABA on the activity of the epidermal ATPase was observed. The data are discussed with respect to the interaction of FC and ABA with the ATPase.     

Studies on high altitude saxicolous-Cyanophyta XI - On the morphology of Petalonema borzii sp. nov.

Abstract

The morphology of a new species of Petalonema Berk collected under a saxicolous habitat at 2900 m above sea level in the Singalila Range, Eastern Himalayas, during the Botanical Expedition of May 1974, was critically studied and named as Petalonema borzii sp. nov. The same alga was collected again on 28 Nov. 1982 from the Singalila Range at an altitude of 2500 m above sea level.     

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.     

Feeding strategy of the sacoglossan opisthobranch <Emphasis Type="Italic">Oxynoe olivacea</Emphasis> on the tropical green alga <Emphasis Type="Italic">Caulerpa taxifolia</Emphasis>

The feeding behaviour of the shelled sacoglossan Oxynoe olivacea was investigated to better understand the role and importance of this species in influencing encroachments of the alien alga Caulerpa taxifolia in the Mediterranean sea. We tested whether this slug preferred, as preliminary field observations suggested, an aggregative feeding behaviour and which part of the algal thallus, phylloid vs rhizoid, it preferred. Results showed that O. olivacea fed in groups and actively selected phylloid. This outcome poses important questions regarding the possibility that this species, fragmenting the alga thallus, could enhance dispersion and regeneration of C. taxifolia.     

Lipid peroxidation in embryos and larvae of the sea urchinStrongylocentrotus intermedius

Lipid peroxidation (LP) and glutathione content were studied at different developmental stages of the sea urchinStrongylocentrotus intermedius: egg cell, fertilization, 4 blastomers, blastula, hatching, gastrula, prism, pluteus. A high rate of LP in the total membrane fraction of sea urchin embryos and larvae at the stages from the egg cell to hatching was observed at enzymatic and nonenzymatic activation of LP. The LP rate was significantly reduced at the gastrula stage and at subsequent stages, there was practically no further development of the process. The glutathione concentration remained unchanged at different stages. The alterations in LP seem to reflect participation of free radicals in regulation of individual development.     

Influence of temperature and salinity on embryonic development ofParacentrotus lividus (Lmk, 1816)

The combined effects of temperature and salinity on early development of the sea urchinParacentrotus lividus (Lmk, 1816) are reported. The optimal temperature-salinity combinations for development are 18 °–20 °C and 34–35‰; there is a significant temperature-salinity interaction. The optimal conditions found in the experiments are above the mean yearly values for the sampled population's environment (North Adriatic Sea), being more similar to those of the Tyrrhenian Sea. These results suggest that embryonic tolerances to temperature and salinity are under genetic and not environmental control.     

Studies on high altitude saxicolous Cyanophyta IV. « Hyphomorpha borzii », sp. nov., a new alga from India

Abstract

The morphology of a new species of Hyphomorpha, Borzi, collected from 3,300 metres above sea level underneath a rock in Singalila Range along Eastern Himalayas, has been studied and named as Hyphomorpha borzii. The relationship, affinities and taxonomy of the alga have been critically discussed at the end.     

Effect of caulerpenyne,a toxin extracted from Caulerpa taxifolia on mechanisms regulating intracellular pH in sea urchin eggs and sea bream hepatocytes

The proliferation of the green marine alga Caulerpa taxifolia in the Mediterranean led us to investigate the toxic effects on marine organisms of caulerpenyne (Cyn), the major secondary metabolite synthesized by the alga. This study was performed on sea urchin eggs (Paracentrotus lividus) and isolated hepatocytes from the sea bream (Sparus aurata), in which accumulation of the toxins by metabolic processes may be of significance. Cyn provoked an acidification of seawater containing both unfertilized and fertilized eggs, as revealed by a titrable efflux of protons. The pHi in unfertilized eggs continuously increased in the presence of Cyn, whereas there was a biphasic response in both fertilized eggs and isolated hepatocytes, with a decrease of the pHi followed by recovery to the initial value. Cyn inhibited the accumulation of ¹⁴C-methylamine in acidic granules present in the cortical zone of sea urchin eggs. Dicyclocarbodiimide (DCCD), a well-known H⁺-ATPase inhibitor, provoked a similar inhibition. Both molecules increased pH in the acidic compartments of isolated bream hepatocytes. These results suggest that Cyn inhibits intracellular sequestration of protons and thus liberates protons into the cell cytoplasm from which they leak toward the extracellular medium. © 1997 John Wiley & Sons, Inc.     

Toxic Prorocentrum lima induces abnormal behaviour in juvenile sea bass

Juveniles of the European sea bass Dicentrarchus labrax were exposed to both cell-free medium and whole cell cultures of the dinoflagellate Prorocentrum lima strain PL2V. Fish were also fed a commercial fish diet in tanks containing live P. lima, and Artemia that had ingested the alga. Fish exposed to the cell-free medium and to whole cell cultures were stressed and behaved abnormally when compared to the behaviour of control fish, fish in normal seawater. Stress-related behaviours included hyperactivities (jumps, fast let-right turns, surface swims, etc), poor feeding reflexes and abstinence from feeding. Fish that directly ingested the alga or that ingested Artemia containing the alga died. Histological studies revealed that gills and liver of treated fish were impacted, as opposed to the normal conditions of same tissues in control fish. The diseased organs could have been responsible for the abnormal behaviours and death of treated fish. The aquaculture and ecological implications of the results are discussed.     

Effect of climatic variation on the relative abundance of the red alga Gelidium robustum in Baja California Sur, Mexico

The red alga Gelidium robustum is important,because of its commercial exploitation in Mexico as araw material for the agar industry, providing 10% ofthe world production of agarophytes. In recent years,its annual harvest in Mexico has shownobvious,variations partly because of an increasedharvesting effort, but also because of environmentalchanges. An analysis is presented of the effect thatinterannual variability of the sea surfacetemperature, wind speed, and upwelling index had onthe relative abundance of this alga from 1980 to 1990.The results indicate a close relation betweenenvironmental fluctuations and the relative abundanceof this species. The response of G. robustum tothe different environmental conditions has not alwaysbeen equal. During El Niño 1982–84, the seasurface temperature was the most important factor andthere was a high negative correlation with therelative abundance. A lag period of three monthsshowed a positive correlation with upwelling index andwind speed. Under normal conditions and during LaNiña, the relative abundance of the alga showed apositive correlation with the sea surface temperature. For the upwelling index and wind speed, therelationship was similar to that during El Niño.     

Effects of free amino acids on the photosynthetic carbon metabolism of symbiotic dinoflagellates

Synthetic host factor (SHF) was used in parallel with crude cnidarian’s host factor (CHF) to investigate their effects on both photosynthetic carbon metabolism and the first step of lipid synthesis in the symbiotic dinoflagellates Symbiodium purchrorum of the sea anemone Aiptasia pulchella. Several species have been studied, namely, sea anemone A. pulchella, reef coral Pocillopora damicornis, and green alga Chlamydomonas reinhardii. Both short-term and long-term experiments with radioactive carbon have shown a higher rate of the alga ¹⁴C photoaccumulation with host factor(s) than of dinoflagellates located in artificial sea water (ASW) alone. In dinoflagellates incubated with both ASW and CHF, ¹⁴C-labeled glycerol was detectable after 15 s of alga illumination. In dinoflagellates isolated from P. damicornis and incubated in CHF and SHF and in dinoflagellates isolated from A. pulchella and incubated in CHF, a higher percentage of ¹⁴C was found in the glycerol as compared to the ASW trial. At the same time, in ASW trial the radioactive label was primarily located in ethanol-soluble lipid fraction. Similar results were observed when dinoflagellates isolated from P. damicornis were incubated with aspartate or glutamate. But there was no effect with taurine, serine, valine, glycine, or lysine. C. reinhardii, incubated in salt-free CHF, partitioned a greater percent of ¹⁴C into the glycerol and less into the ethanol-soluble lipids as compared to the corresponding control incubations. The amount of ¹⁴C in neutral and polar lipids was identical in that in A. pulchella dinoflagellates incubated in ASW or CHF. The arrays of neutral ¹⁴C-lipids produced under both ASW alone and ASW with CHF conditions, and over time were not significantly distinguishable. Host factors appeared to provide an optimum environment to sustain maximum metabolic efficiency. A biochemical model, based on the quantitative and qualitative assessment of carbon pathways in dinoflagellates incubated both in host factor and sea water alone, is presented. Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 2, pp. 195–208. The text was submitted by the authors in English.