CLONAL VARIATION IN PHOTOTAXIS AND SETTLEMENT BEHAVIORS OF HINCKSIA IRREGULARIS (PHAEOPHYCEAE) SPORES1

Hincksia irregularis (Kützing) Amsler isolates from North Carolina were used to assess clonal variations in spore swimming behavior and settlement responses within a population. These behaviors were also compared with those previously reported from a Florida panhandle clone to assess differences between geographically distinct populations. Minor variation between morphologies, reproductive responses, and settlement intervals were observed between all clones. Spores from all North Carolina clones exhibited positive phototaxis, but there was significant variation in the lower threshold irradiances that could elicit the behavior. In contrast, spores from the previously studied Florida panhandle clone exhibited negative phototaxis. Hincksia irregularis spore settlement responses to hydrophobic and hydrophilic surfaces were consistent for all clones currently and previously tested. Interactive effects of light and charge on settlement were evident in spores from North Carolina isolates, but these effects differed from the Florida panhandle isolate. Settlement preference and swimming behavior differences may have a considerable effect on an individual's ecological fitness and placement within the marine environment. However, the perceived relevance of these differences needs to be validated further in light of previously reported genetic, physiological, and behavioral variations observed in other algal clones that have called into question traditional assessments of individuality.     

NUTRIENTS DO NOT INFLUENCE SWIMMING BEHAVIOR OR SETTLEMENT RATES OF ECTOCARPUS SILICULOSUS (PHAEOPHYCEAE) SPORES

Spores newly released from plurilocular sporangia of Ectocarpus siliculosus (Dillw.) Lyngb. sporophytes were assayed for chemotaxis to nutrients and for settlement stimulation by nutrients. To enable these measurements with relatively small volumes and numbers of released spores, we developed a computer-assisted motion-analysis assay for spore chemotaxis and verified the results with a more standard, capillary tube chemotaxis assay. The presence of a nutrient gradient did not influence the swimming behavior of E. siliculosus spores in the motion-analysis assay, and likewise no chemotactic effect was measured in the capillary tube assay. Microplate settlement assays previously utilized with bacteria and invertebrates were adapted for use with algal spores. E. siliculosus spores settled at higher rates on a hydrophobic plastic surface than on surfaces with either positively or negatively charged hydrophilic coatings. Nutrient mixtures had no effect on the rate of spore settlement on hydrophobic surfaces.     

A NOVEL BIOFOULING ASSAY USING COMPUTER-ASSISTED MOTION ANALYSIS OF HINCKSIA IRREGULARIS SPORE SWIMMING

Chloroplasts in heterokont algae probably originated from a red algal endosymbiont which was engulfed and retained by a eukaryotic host, and are surrounded by four envelope membranes. The outermost of these membranes is called chloroplast ER (CER) and usually connects with the nuclear envelope. This information, however, is based mainly on studies on single-plastid heterokont algae. In multi-plastid heterokont algae, it is still unclear whether CER is continuous with the nuclear envelope. Since nuclear-encoded chloroplast proteins are synthesized by ribosomes on the ER membrane, clarifying the ER-CER structure in the heterokont algae is important in order to know the targeting pathway of those proteins. We did a detailed ultrastructural observation of endomembrane systems in a multi-plastid heterokont alga: Heterosigma akashiwo , and confirmed that the CER membrane was continuous with the ER membrane. However, unlike the CER membranes in other heterokont algae, it seemed to have very few ribosome attached. We also performed experiments for protein targeting into canine microsomes using a precursor for a nuclear-encoded chloroplast protein, a fucoxanthin-chlorophyll protein (FCP), of H. akashiwo , to see if the protein is targeted to the ER. It demonstrated that the precursor has a functional signal sequence for ER targeting, and is co-translationally translocated into the microsomes. Based on these data, we propose a hypothesis that, in H. akashiwo , nuclear-encoded chloroplast protein precursors that have been co-translationally inserted into the ER lumen are sorted in the ER and transported to the chloroplasts through the ER.     

LIGHT BOUNDARIES AND THE COUPLED EFFECTS OF SURFACE HYDROPHOBICITY AND LIGHT ON SPORE SETTLEMENT IN THE BROWN ALGA HINCKSIA IRREGULARIS (PHAEOPHYCEAE) 1

We examined the effects of light and surface hydrophobicity individually and in tandem on Hincksia irregularis (Kützing) Amsler spore settlement. Hincksia irregularis spores were determined to be negatively phototactic by the use of computer‐assisted motion analysis. Spore settlement was significantly influenced by surface hydrophobicity and by light, individually and in tandem. Experiments conducted using culture plates modified to reduce well edge artifacts revealed significantly higher settlement on hydrophobic surfaces and in dark environments when compared with negatively charged surfaces and lighted environments. Experimental light/dark boundaries elicited distinct spore settlement responses, with spores displaying dissimilar settlement patterns on plates with different surface hydrophobicities. The results of this study indicate H. irregularis spores possess the capacity for complex responses to their environment. These complex responses may influence dispersal and aid spores in the detection of suitable settlement locations in marine microenvironments.     

SURFACE INTERACTIONS OF THE EPIPHYTIC MACROALGA HINCKSIA MITCHELLIAE (PHAEOPHYCEAE) WITH THE SHOALGRASS HALODULE WRIGHTII (CYMODOCEACEAE)1

Meadows of Halodule wrightii (Cymodoceaceae) underwent a decline in a tidal flat located at Paranaguá Bay (Parana, SE Brazil). This decline appeared to be related to an overgrowth of the epiphytic macroalga Hincksia mitchelliae (Harv.) P. C. Silva (Phaeophyceae). In order to characterize the type of epiphytism between the alga and its plant host, we compared two samples from the beginning and end of the algal overgrowth via electron and optical microscopes. The investigation revealed that at both sampling periods, there was an epiphytism of type II, which is due to an infection of epiphytes strongly attached to the surface of the host but not associated to any apparent direct host‐tissue damage. The presence of plasmodesmata between the cells of Hincksia only in the late stage of the host–epiphyte interaction indicated a change in the vegetative organization of Hincksia in relation to its host to improve nutrient absorption and distribution through the epiphyte cells. This is the first report on plasmodesmata in H. mitchelliae. The proposed mechanisms with which the algal epiphytes lead seagrasses to death are shadowing by adhesion on Halodule surface and disruption of its osmoregulatory system. Our findings have implications for the conservation and management strategies of seagrass ecosystems.     

A METHOD FOR THE QUANTITATIVE AND QUALITATIVE DETERMINATION OF PLANKTONIC DIATOMS

SUMMARY

A method for the quantitative and qualitative determination of planktonic diatoms was developed. The method uses the Utermöhl counting technique (in which an inverted microscope is employed) as a basis but also involves the calculation of the relative density of each species in the association (as determined with the ordinary light microscope). A combination of the results thus obtained ensures that even the smallest diatoms (2,5 μm) are accounted for. These often constitute a significant proportion of the population, but are often overlooked or may not even be resolved using Utermöhl's method.     

CRYOANALYTICAL STUDIES OF FREEZING DAMAGE AND RECOVERY IN FUCUS VESICULOSUS (PHAEOPHYCEAE)

Receptacles of Fucus vesiculosus L. were examined using cryoscanning electron microscopy of rapidly frozen, hydrated tissue to avoid extraction and collapse of the extracellular matrix (ECM) and provide new structural information. The pore of the conceptacle was occluded by the tightly appressed tips of the paraphyses. These were embedded in copious ECM within the conceptacle. The ECM inside the conceptacle and in the medulla differed markedly in texture and with respect to the distribution of S and Cl, as revealed by energy-dispersive X-ray microanalysis. High levels of S were restricted to the conceptacle's ECM, whereas the ECM in the medulla was always S poor and Cl rich. Receptacles frozen naturally at low tide during the winter of 1998 showed extensive damage in the medulla, and this likely accounts for the cessation of reproduction during the winter on the Maine shore. The damaged receptacles recovered and resumed growth during the spring, and they released gametes by May. Abscission of the overwintered receptacles was observed in June to July, coincident with the formation of new receptacles at the tips of newly grown portions of the fronds.     

PHOTOSYNTHETIC PHYSIOLOGY AND CHEMICAL COMPOSITION OF SPORES OF THE KELPS MACROCYSTIS PYRIFERA,NEREOCYSTIS LUETKEANA,LAMINARIA FARLOWII,AND PTERYGOPHORA CALIFORNICA (PHAEOPHYCEAE)1

Recently released spores of the kelps Macrocystis pyrifera (L.) C. Ag., Nereocystis luetkeana (Mert.) Post. and Rupr., Laminaria farlowii Setch., and Pterygophora californica Rupr. had different levels of net photosynthesis. Spore-specific photosynthesis–irradiance relationships were similar in many respects for M. pyrifera, N. luetkeana, and L. farlowii spores. All three species had low rates of net light-saturated photosynthesis. In contrast, spores of P. californica had higher photosynthetic potential and overall net photosynthesis than the other three species. On a cell carbon basis, however, photosynthetic rates in N. luetkeana spores were similar to those of P. californica spores and higher than those of M. pyrifera spores. Chlorophyll a content of spores varied 10-fold among species. The rank order of significant differences in chlorophyll a content was P. californica > L. farlowii > N. luetkeana > M. pyrifera. As a result, chlorophyll-specific measurements suggest M. pyrifera and N. luetkeana spores had much higher quantum efficiency and photosynthetic potential than either P. californica or L. farlowii spores. Maternal carbon and nitrogen investment significantly differed in spores of M. pyrifera, N. luetkeana, and P. californica with P. californica > M. pyrifera > N. luetkeana. Carbon content in spores of each of these three species increased by about 30% during 12 h of saturating irradiance. We suggest that the photosynthetic capabilities of and maternal investment in spores may be related to the spore as a unit of dispersal, to the reproductive ecology of the parental sporophytic stages, and to the growth and physiology of the germling gametophyte stages.     

EXPERIMENTAL MODIFICATION OF THE MORPHOGENETIC BEHAVIOR OF THE ISOLATED SUB-APICAL CELL OF THE APEX OF SPHACELARIA CIRROSA (PHAEOPHYCEAE)1

Experimental isolation of the terminal cells of the apex of Sphacelaria cirrosa (Roth) C. Agardh shows that the isolated apical cell retains a self-maintained mode of functioning. The isolated sub-apical cell, however, gives rise to a newly-formed axis after undergoing a different type of developmental sequence. An experimental scheme allowing one to defer isolation of the sub-apical cell after killing the apical cell shows that the maintenance, for a few hours only, of connections with subjacent cells is sufficient to induce an apical type mode of functioning in the sub-apical cell. The sequential analysis of the first cytological events in this phenomenon allows one to relate this modification of the morphogenetic program to a functional and polarized arrangement of organelles within the cell reconstituting the apical. The morphogenetic program is characterized by a migration of the nucleus to a distal position prior to an asymmetrical mitosis characteristic of the apical mode of functioning.     

QUANTITATIVE AND QUALITATIVE DIFFERENCES IN PLANT RESPONSE TO THE GIBBERELLINS

Wittwer , S. H., and M. J. Bukovac . (Michigan State U., E. Lansing.) Quantitative and qualitative differences in plant response to the gibberellins. Amer. Jour. Bot. 49(5): 524–529. Illus. 1962.—The comparative biological activities of gibberellins A₁ through A₉ were evaluated, over a wide concentration range and in several test systems. All gibberellins were effective in promoting stem elongation of dwarf peas (Pisum sativum), and, with the exception of A₈, epicotyl growth in Phaseolus vulgaris. Elongation of Cucumis sativus seedlings was strikingly greater with A₄, A₇, and A₉ than with the other gibberellins. With mutant dwarfs of Zea mays, A₅ and A₉ were the most active gibberellins for d₃ and d₅, and relatively ineffective compared to A₃ on d₁. Gibberellins A₂, A₇, and A₈ were less effective than A₃ on all dwarfs. Qualitative and quantitative differences among the gibberellins were noted on seedstalk elongation and flowering of Lactuca sativa, with A₃ the most active followed by A₁, A₇, A₄, and A₉. No flowering or seedstalk elongation occurred with A₂, A₆ or A₈. Parthenocarpic fruit growth in Lycopersicon esculentum was a function of dosage with all gibberellins. At the lowest levels, A₅ and A₇ were the most active, while at the highest levels all gibberellins with the exception of A₈ were equally effective. The results suggest a high degree of species and response specificity among the known fungal and higher plant gibberellins, and demonstrate the importance of utilizing a wide spectrum of plant responses and dosage levels in the biological assay of plant extracts for native gibberellins.     

QUANTITATIVE AND QUALITATIVE ANALYSES OF PROTEIN SYNTHESIS DURING HEAT SHOCK IN THE MARINE DIATOM NITZSCHIA ALBA (BACILLARIOPHYCEAE)1

Protein synthesis in the diatom Nitzschia alba Lewin and Lewin was drastically altered when the cells were incubated at a supraoptimal temperaeture. Quantitatively, the overall protein synthesis was greatly suppressed as indicated by teh rate of [³⁵S] methionine incorporation. The extent of suppression of protein synthesis was proportional to the severity of the heat-shock treatment which was a combination of elevated temperature and treatment duration. The in viro synthesized proteins were also qualitativelty anlayzed by two-dimensional gel electrophoresis. Dependeing on the treatment condition, a set of heat-shock proteins (HSPs) were induced. They were best detected when the cells were subjected to shocks of 35°C for 60 min or 40°C for 10 min followed by a 60 min labelling at 30°C. The results revealed 16 HSps which had moluecular weights ranging from 24–94 kD and isoelectric points ranging from 5.50–7.10. Some of the HSps were identical in molelcular weights but with differeentr isoelectric points. The induction and accumulation of HSPs in Nitzschia alba were transitory. Prologned heat-shock treatments resulted in a complete cessation of protein syntehsis and no further induction of HSPs. In all aspects, the heat shock response of diatoms was similar to that in higher plants such as soybean, maize and tobacco but differenet from most animal systems.     

QUANTITATIVE GENETICS OF SPRINT RUNNING SPEED AND SWIMMING ENDURANCE IN LABORATORY HOUSE MICE (MUS DOMESTICUS)

We tested the hypothesis that locomotor speed and endurance show a negative genetic correlation using a genetically variable laboratory strain of house mice (Hsd:ICR: Mus domesticus). A negative genetic correlation would qualify as an evolutionary “constraint,” because both aspects of locomotor performance are generally expected to be under positive directional selection in wild populations. We also tested whether speed or endurance showed any genetic correlation with body mass. For all traits, residuals from multiple regression equations were computed to remove effects of possible confounding variables such as age at testing, measurement block, observer, and sex. Estimates of quantitative genetic parameters were then obtained using Shaw's (1987) restricted maximum-likelihood programs, modified to account for our breeding design, which incorporated cross-fostering. Both speed and endurance were measured on two consecutive trial days, and both were repeatable. We initially analyzed performances on each trial day and the maximal value. For endurance, the three estimates of narrow-sense heritabilities ranged from 0.17 to 0.33 (full ADCE model), and some were statistically significantly different from zero using likelihood ratio tests. The heritability estimate for sprint speed measured on trial day 1 was 0.17, but negative for all other measures. Moreover, the additive genetic covariance between speeds measured on the two days was near zero, indicating that the two measures are to some extent different traits. The additive genetic covariance between speed on trial day 1 and any of the four measures of endurance was negative, large, and always statistically significant. None of the measures of speed or endurance was significantly genetically correlated with body mass. Thus, we predict that artificial selection for increased locomotor speed in these mice would result in a decrease in endurance, but no change in body mass. Such experiments could lead to a better understanding of the physiological mechanisms leading to trade-offs in aspects of locomotor abilities.     

THE SANITATION OF FISH BOXES. I. THE QUANTITATIVE AND QUALITATIVE BACTERIOLOGY OF COMMERCIAL WOODEN FISH BOXES

SUMMARY: The cleaning efficiency of hosing wooden market fish boxes with cold water, the usual commercial practice, was very low, even when high pressure jets were used. After washing, the internal surfaces could still carry up to 20 × 10⁶ bacteria/cm². Even prolonged steaming failed to sterilize the surfaces. Coryneform organisms accounted for over 50% of the total bacterial flora, although these have usually been reported as present in only low proportions on both fresh and spoiling fish. The major subsidiary groups were species of Achromobacter and Pseudomonas , constituting about 18% and 14% of the population respectively. A few micrococci and flavobacteria were also present.
The conditions of counting, i.e. whether roll tubes or Petri dishes were used, whether incubation was at 0°, 20° or 37°, and whether the media were based on sea or tap water, could markedly influence both the magnitude of the count and the proportions of the different types of bacteria which could be isolated.     

THE NORMAL INTESTINAL FLORA OF THE PIG. III. QUALITATIVE STUDIES OF LACTOBACILLI AND STREPTOCOCCI

SUMMARY: Streptococci and lactobacilli were isolated from the faeces and caecal contents of experimental pigs receiving various diets. Fifty-seven strains of streptococci and forty-one (representative of 220) lactobacilli were examined physiologically and serologically. Lactobacillus acidophilus and L. fermenti predominated among the lactobacilli, while the largest group of streptococci resembled the 'unclassified' strains of Barnes & Ingram (1955) and Barnes, Ingram & Ingram (1956) from bacon factory premises, pig faeces and canned hams.     

THE INFLUENCE OF pH AND LICHEN METABOLITES (VULPINIC ACID AND (+) USNIC ACID) ON THE GROWTH OF THE LICHEN PHOTOBIONTTREBOUXIA IRREGULARIS

The quantitative response to two lichen metabolites, vulpinic acid and (+) usnic acid, on biomass production by the photobiontTrebouxia irregularis, isolated from the lichenCladina mitis, was determined experimentally. The presence of usnic acid resulted in inhibition of photobiont growth, and the effect depended on the pH of the culture medium. The concentration effect of usnic acid was observed. The application of vulpinic acid almost completely inhibited the growth ofT. irregularisand no significant differences were found among samples at different medium pH values.