THE APICAL MERISTEM OF SPLACHNIDIUM RUGOSUM (PHAEOPHYTA)1

The meristem of Splachnidium rugosum consists of a central apical cell surrounded by a region of actively dividing cells, many of which bear hairs. Conceptacle initials are scattered throughout the surface layer of the meristematic region. Conceptacle initials and apical hairs differentiate adjacent to the apical cell. The apical cell and the conceptacle initials are distinctive, pear-shaped cells possessing similar cytological features that are consistent with significant metabolic activity. They have a nucleus surrounded by dictyosomes, a stellate chloroplast, mitochondria, and numerous vesicles and physodes. When the apical cell is damaged as a result of experimental manipulation, growth ceases. It is inferred that the apical cell controls cell division in the meristematic region and also the differentiation of conceptacle initials and apical hairs. The apical meristems of Splachnidium and species of the Fucales have several important features in common, including the growth-regulatory role of the apical cell and the process of conceptacle initiation. The taxa may possibly have a common evolutionary origin. The problematic and unresolved taxonomic status of Splachnidium is discussed.     

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.     

ULTRASTRUCTURE OF VEGETATIVE AND DIVIDING CELLS OF THE UNICELLULAR RED ALGAE RHODELLA VIOLACEA AND RHODELLA MACULATA1

Rhodella violacea (Kornmann) Wehrmeyer and Rhodella maculata Evans were investigated for ultrastructural details of vegetative and dividing cells. Rhodella violacea has a nuclear projection into the pyrenoid similar to that found in R. maculata, although the nuclear projection in R. maculata traverses a starch-lined area before contacting the pyrenoid. Unlike most, red algae, the two Rhodella species lack a peripheral encircling thylakoid in the chloroplast and have dictyosomes associated solely with endoplasmic reticulum (ER) instead of with both mitochondria and ER. Both species also have a well-developed peripheral system of ER connected to the plasmalemma by tubules, a situation found only in red algal unicells, Cell division was studied primarily in R. violacea; a less thorough examination of R. maculata showed no essential differences. Both have small, double-ringed, nucleus-associated organ files (NAOs) surrounded by moderately electron-dense material, metaphase–anaphase polar gaps in the nuclear envelope, absence of perinuclear ER. and short interzonal spindles. This pattern of mitosis is similar in most respects to that reported in the unicell Flintiella. Following mitosis, microtubules extend from the region of each NAO to its associated nucleus and to the undivided pyrenoid. The NAOs appear to apply tension to the nuclei and the pyrenoid and may be the mechanism for ensuring equal partitioning of both organdies. Two different forms of pyrenoid-nucleus association occur during mitosis. Nuclear projections into the pyrenoid, prevalent during interphase and early stages of mitosis, recede at metaphase. Then, the pyrenoid extends protrusions into the nuclear polar areas, forming a cup that partially surrounds the nucleus. Cell division and vegetative characters confirm the close taxonomic affinity of these two species of Rhodella and support their separation from the genus Porphyridium.     

DARK CARBON FIXATION STUDIES ON THE INTERTIDAL MACROALGA ASCOPHYLLUM NODOSUM (PHAEOPHYTA) 1

The characteristics of dark carbon fixation by Ascophyllum nodosum were investigated. In longitudinal profile the maximum rates of dark and light dependent fixation are found at the apex. The use of Michaelis-Menten kinetics did not suitably describe the relationship between the uptake rate in the dark and the total inorganic carbon concentration. Dark fixation was saturated at a total inorganic carbon concentration [TIC] of 2.5 mM. The use of the Hill-Whittingham equation to describe the uptake curve indicates that the process is diffusion limited. Comparisons of dark fixation at high (8.0) and low (5.2) pH suggest that bicarbonate ions are used as a source of inorganic carbon. The transfer of ¹⁴C, fixed in the dark, from the ethanol soluble to the insoluble fraction was relatively slow irrespective of the light treatment during the chase period. Ascophyllum nodosum displays a small diel fluctuation in the pH of aqueous extracts and titratable acidity similar to that displayed by CAM plants. The significance of dark fixation to the overall carbon budget is discussed.     

SEQUENCE AND PHYLOGENY OF THE PSAB GENE OF PYLAIELLA LITTORALIS(PHAEOPHYTA)1

The psaB gene codes for one of two highly conserved P700 chlorophyll a apoproteins of photosystem I. This gene was cloned from the brown alga, Pylaiella littoralis (L.) Kjellm., and its primary sequence was determined. The inferred amino acid sequence of the P. littoralis protein was compared to homologous sequences from land plants, green algae, and a cyanobacterium. The psaB protein sequence is very conserved in all the examined taxa, and an unrooted phylogenetic tree, generated from a distance matrix, shows that the P. littoralis gene is closer to that of the cyanobacterium Synechocossus sp. PCC 7002 than are those of green algae, land plants, and Euglena gracilis.     

TAXOL REDUCES THE RATE OF CYTOKINESIS IN PtK1CELLS

Mitotic PtK₁cells were treated both during mid-anaphase and at furrow initiation with the potent microtubule (MT) stabilizing agent, taxol, to determine the role of MTs in the rate of cytokinetic events. Rates of cytokinesis (μm/min) were measured by changes in furrow diameter. Incubation of PtK₁cells during mid-anaphase with 5 μg/ml taxol slows the rate of cytokinesis by an average of 43%. Instead of furrow initiation to midbody formation taking an average of 10.7 min (1.6 μm/min), furrowing to midbody formation was completed in an average of 19.0 min (0.9 μm/min), which does not include the 7-min period between taxol application in mid-anaphase and furrow initiation. Application of 5 μg/ml taxol to cells at furrow initiation had a reduced effect on decreasing the rate of cytokinesis and midbody formation; furrowing to midbody formation took an average of 14.6 min (1.2 μm/min). These data suggest that delays in the rate of cytokinesis is dependent on the mitotic stage at which taxol is applied. Ultrastructural analysis shows that taxol treatment of anaphase cells prevents midbody formation during early G₁, yet MT number and organization in the furrowed region is not significantly altered from untreated cells. There is little change in the organization and amount of contractile ring microfilaments, yet filaments are also found parallel to midbody MTs. Our results may be explained by the fact that taxol tends to stabilize MTs which probably affects the rate at which they depolymerize in the terminal phases of cytokinesis. Reduction in depolymerization rates of a stable population of MTs could serve to regulate the rate of cytokinesis.     

FIELD AND CULTURE STUDIES OF A POPULATION OF ENDARACHNE BINGHAMIAE (PHAEOPHYTA) FROM SOUTHERN CALIFORNIA1

Abundances of the erect, blade phase of Endarachne binghamiae J. Ag. (Scytosiphonales, Phaeophyta) varied seasonally at a southern California rocky intertidal site. Blade cover and density were much greater in the fall through early spring; blades were mostly absent from quadrats during the summer. Blade abundances were negatively correlated with both seasonal variations in seawater temperature and photoperiod. Laboratory culture studies failed to provide evidence for sexual reproduction. The life history appears to be of the “direct” type with plurangia-produced zooids germinating into crustose disks. Most disks developed erect blade clusters under spring/fall (17° C) and winter (13° C) temperatures over the range of natural photoperiods employed (14:10, 12:12, 10:14 h LD). In contrast, cultures held under the summer temperature (21° C) produced almost entirely crustose growths regardless of photoperiod. Similar results were obtained for cultures grown at 100 and 200 μE · m^?2· s^?1. E. binghamiae blades were fertile throughout the year and produced viable zooids indicating that reproductive seasonality did not influence the seasonal pattern of blade abundance. Culture and field studies suggest that the initiation of new erect blade clusters from crustose disks is confined to the cooler months of the year (winter and spring). The summer reduction or absence of E. binghamiae blades appears to be due to increased mortality rates and temperature constraints on the development of new erect bladed thalli. Hypothetical causes of mortality are desiccation stress, sand burial, increased grazing activity and a genetically-based short life span.     

TRANSLOCATION OF IODINE IN LAMINARIA SACCHARINA(PHAEOPHYTA)1

Long-distance translocation of ¹²⁵I in Laminaria saccharina (L.) Lamour. followed a “source to sink” pattern. When the source of ¹²⁵I was placed on the distal mature part of the blade, the translocation was unidirectional, basipetal and directed towards the meristematic region at the blade-stipe junction. When the source was placed directly at the meristem there was no movement of label distal to the meristem. The velocity of¹²⁵I transport ranged from 2 to 3.5 cm · h^?1. The anion I^? seemed to be the only species of¹²⁵I transported. An assay of iodine content in different parts of L. saccharina plant showed much higher levels of iodine in the meristem, stipe and holdfast than in the blade. This distribution concurs well with the pattern of I^? translocation.     

BIOMECHANICS AND ALGINIC ACID COMPOSITION DURING HYDRODYNAMIC ADAPTATION BY EGREGIA MENZIESII (PHAEOPHYTA) JUVENILES1

To investigate the possible link between cell wall alginic acid composition and tissue mechanics, juveniles of Egregia menziesii (Turn.) Aresch. were grown under controlled conditions in an outdoor flowing seawater system under three different force regimes. After 6–10 weeks of growth, tissue strength, breaking strain, modulus, toughness, work of fracture, and the percentage of polymannuronate, polyguluronate, and alternating sequences in the cell wall alginic acid were examined. The force regime had significant effects on all mechanical indices except toughness. Juveniles grown under high energy conditions (water velocity = 1.2 m · s^?1) were about two times stronger, two times stiffer, and had a 1.5 times greater work of fracture than those from low energy conditions (<1 cm ·^?1). Treatment effects on thallus strength and modulus were predicted from alginic acid composition data to test for the importance of this cell wall material in whole plant adaptation to hydrodynamic stress. However, the prediction that differences in alginic acid composition were responsible for differences in tissue mechanical properties was inconsistent with observations. Therefore, the hypothesis that alginates play a central role in structural adaptation could not be accepted.     

EFFECTS OF WOUNDING BY THE HERBIVOROUS SNAILS LITTORINA SITKANA AND L. SCUTULATA (MOLLUSCA) ON GROWTH AND REPRODUCTION OF THE INTERTIDAL ALGA FUCUS DISTICHUS (PHAEOPHYTA)1

Small (3–7 cm long) Fucus distichus ssp. edentatus (de la Pyl.) Pow. Plants were tagged at three sites in which densities of the herbivorous snails Littorina sitkana and L. scutulata ranged from 367–4690 animals · M^-2. From April–August 1986, the growth rate, degree of wounding, and reproductive status of individual thalli were monitored at 2–4 week intervals. Grazer-inflicted damage to the thalli varied within and among sites. Mean growth rates at the site with low densities of littorines were about twice those at the site with intermediate densities and about four times those at the high density site. At the site with high densities of littorines, F. distichus growth rates were negatively correlated with the degree to which the plants were wounded. There appeared to be no correlation of grazer density with F. distichus survivorship. Thalli at the site with few herbivores tended to reproduce earlier and at a larger size than did those at the other two sites. In all three areas, only thalli that had received little damage from herbivores became reproductive. By lowering growth rates and delaying reproduction in F. distichus, grazing by littorine snails can potentially cause variation in reproductive output among individual thalli.     

FINE STRUCTURE AND HISTOCHEMISTRY OF VESICLE CELLS OF THE RED ALGA ANTITHAMNION DEFECTUM (CERAMIACEAE)1

The ultrastructure and histochemistry of the refractile, vesiculate cells (“blasenzellen,”“cellules secretrices,”“gland cells”) of Antithamnion defectum Kylin were examined. The refringent vacuolar contents disclosed two components of differing density: an electron opaque, proteinaceous matrix material surrounding cores of irregularly shaped, less opaque material. The cores contain less protein and more unknown material than the matrix. Part or all of the vacuolar material is synthesized by abundant rough endoplasmic reticulum (ER) and deposited in smooth surfaced cisternae that swell to form vesicles. Mitochondria are usually associated with stacks of the swelling cisternae. The vesicles enlarge by continued deposition of synthesized material and coalescence with other vesicles. All vesicles eventually coalesce to form the mature vacuole. A crystalline array of fibrils develops in the cytoplasm during later stages of vacuole enlargement. The crystal contains a sulfated, acidic polysaccharidic material. The chloroplasts, if present, and nucleus degenerate at vacuole maturity. Active release of the vacuolar material does not occur, and organelles for extracellular secretion are not present. Structural evidence suggests a storage, rather than secretory, function for the cells.     

MITOSIS IN THE OCTAFLAGELLATE PRASINOPHYTE,PYRAMIMONAS AMYLIFERA (CHLOROPHYTA)1

Cell division is described in the octaflagellate prasinophyte Pyramimonas amylifera Conrad and is compared in related genera. Basal bodies replicate at preprophase and move toward the poles. Cells remain motile throughout division. The nuclear envelope disperses and chromosomes begin to condense at prophase. Pairs of multilayered kinetochores are evident on the chromosomes of the metaphase plate. Spindle microtubules extending from the region of the basal bodies and rhizoplasts attach to the kinetochores or extend from pole to pole. Numerous vesicles and ribosomes have entered the nuclear region and the incipient cleavage furrow invaginates. The chromosomes move toward the poles at anaphase leaving a broad interzonal spindle between the two chromosomal plates. The nuclear envelope reforms first around the chromatin on the side adjacent to the spindle poles and later on the interzonal side. The cleavage furrow progresses into the interzonal spindle at telophase. By late telophase the nucleoli have reformed and the chromosomes have decondensed. The interzonal spindle has not been observed late in telophase. As the cleavage furrow nears completion the cells begin to twist and contort, ultimately separating the two cells.     

KARYOLOGICAL STUDIES OF THREE SPECIES OF SCYTOSIPHONACEAE (PHAEOPHYTA) FROM COASTAL NORTH CAROLINA1

Chromosome numbers for three species of Scytosiphonaceae from the warm temperate coast of North Carolina are presented. Petalonia fascia (O. F. Mueller) Kuntze, P. zosterifolia (Reinke) Kuntze, and Scytosiphon lomentaria (Lyngbye) C. Agardh were all found to have twenty-two chromosomes in both macro- and microthallus stages. The uniformity of chromosome numbers and morphology for all three taxa suggest a close genetic relationship and possible common ancestry. Evidence of synchronous mitotic divisions is presented for the first time in the Phaeophyta. Both field collected and culture material of P. fascia and P. zosterifolia exhibited a 2–4 h post-sunset peak in nuclear divisions. In actively growing macrothalli, up to 8% of the observed cells were found in some stage of mitosis. Data suggest that mitosis and the entire sequence of cytokinesis require approximately 2 h.     

EFFECT OF HIGH IRRADIANCE ON RECRUITMENT OF THE GIANT KELP MACROCYSTIS (PHAEOPHYTA) IN SHALLOW WATER1

Laboratory and field experiments were done hi Still-water Cove, Carmel Bay, California, and Monterey Harbor, California, to determine the effect of photosynthetically active radiation (PAR) on the shallow (upper) limit of giant kelp, Macrocystis pyrifera (L.) C. Agardh. At shallow depths, M. pyrifera did not recruit or grow to macroscopic size from gametophytes or embryonic sporophytes transplanted to vertical buoy lines; sharp decreases in PAR with depth coincided with observed recruitment and sporophyte distributions. Shade manipulations indicated that settlement of M. pyrifera zoospores was decreased, but not prohibited, by high PAR. Postsettlement stages (gametophytes and embryonic sporophytes), however, survived only under shade. These results suggest that high PAR can inhibit the recruitment of M. pyrifera to shallow water by killing its postsettlement stages; whether or not ultraviolet (UV) radiation also inhibits recruitment was not tested. In either case, however, it appears that high irradiance (PAR and/or UV) regulates the shallow limit of M. pyrifera prior to temperature and desiccation stresses inherent to intertidal regions. In an additional experiment, recruitment or growth of transplanted gametophytes or embryonic sporophytes of Macrocystis integrifolia Bory also did not occur at shallow depths, suggesting that this shallow water species accesses high irradiance regions via a method other than sexual reproduction.     

EFFECTS OF MICROTUBULE INHIBITORS ON PRONUCLEAR MIGRATION AND EMBRYOGENESIS IN FUCUS DISTICHUS(PHAEOPHYTA)1

Pronuclear migration in Fucus distichus spp. edentatus (de la Pyl.) Powell is blocked by incubation of fertilized eggs in colchicine (1 mg/ml) and Nocodazole (2 μg/ ml). Rhizoids form prior to decondensation of the sperm chromatin in eggs in which pronuclear fusion is blocked. This occurs during continuous colchicine incubation as well as in eggs recovering from a short treatment with either drug following fertilization. During recovery of the cells, the sperm and egg chromosomes condense, and the sperm chromosomes migrate toward the egg pronucleus. The delay in migration following removal of colchicine is as much as 24 h and is even slower following removal of Nocodazole. The egg chromosomes form a metaphase plate in treated cells while the sperm chromosomes are still distant in the cytoplasm. This suggests that egg centrioles are important in the mitotic division of the zygote, not sperm centrioles. The effect of colchicine treatment on the mitotic plane and cytokinesis is also discussed.     

THE OCCURRENCE OF FLAGELLATED EGGS IN LAMINARIA ANGUSTATA (PHAEOPHYTA,LAMINARIALES)1

Eggs of laminaria angustata Kjellman were shown to have two flagella. Compared with flagella of other phaeophycean swarmers, those of Laminaria eggs have several unique characters such as lack of mastigonemes, widely spaced basal bodies and no flagellar rootlets. The flagella abscise during egg liberation.     

AN ELECTRON MICROSCOPE STUDY OF GAMETE RELEASE AND SETTLING IN THE COMPLANATE FORM OF SCYTOSIPHON (SCYTOSIPHONACEAE,PHAEOPHYTA)1

This paper describes the ultrastructural characteristics of gametes and their liberation from the gametangia in Scytosiphon sp., a brown alga showing only slight sexual differentiation. Both male and female gametes are released initially into a central cavity which forms in the gametangial sorus by the extensive dissolution of the internal cell walls. Scytosiphon sp. gametes possess structural features in common with the zoids of other species of brown algae. Gamete fine structure is discussed in relation to cell function. After release from the gametangial sorus, female gametes can be distinguished from males by the presence of a large number of Golgi-derived vesicles with electron dense cores. It is possible that these vesicles contain the sex attractant compound. When gametes settle they become spherical, the flagella and eyespot are withdrawn into the cell and adhesive material, apparently originating from the activity of the Golgi body, appears on the surface of the cell.     

OCCURRENCE OF GLYCOLATE OXIDASE AND HYDROXYPYRUVATE REDUCTASE IN EGREGIA MENZIESII (PHAEOPHYTA)1

Glycolate oxidase and hydroxypyruvate reductase, two key enzymes of glycolate metabolism, are present in the brown alga Egregia menziesii (Turn.) Aresch. The pH optimum and Km for the partially purified glycolate oxidase are similar to the enzyme from higher plants, Charophyta and Xanthophyta, whereas the substrate specificity is different from the higher plant enzyme.     

STUDIES ON THE FINE STRUCTURE OF ULTRACENTRIFUGED SPINAL GANGLION CELLS

The following structures were observed in electron micrographs of the mouse spinal ganglion cells: Nissl bodies composed of both aggregated rough-type, largely oriented, membranes of the endoplasmic reticulum and discrete particles; short rodlike mitochondria with well-developed transverse, obliquely or longitudinally arranged cristae, and a relatively typical Golgi complex. The components of ultracentrifuged ganglion cells (400,000 times gravity for 20 minutes) are stratified, the layers appearing in the order of their decreasing density as follows: (1) A microsomal or ergastoplasmic layer which may be further divided into three sublayers without sharp boundaries, namely, a discrete particle layer, a layer of discrete particles and highly distorted membranes of the endoplasmic reticulum, and a layer composed of relatively intact, but stretched membranes of the endoplasmic reticulum and discrete particles. (2) Mitochondria constitute a relatively broad layer. They are sometimes stretched; however, they retain most of their fine structure. The stratified nucleus is found within the mitochondrial layer. (3) A relatively wide layer of tightly packed vesicles. (4) At the centripetal end, resting against the cell membrane, are a few lipid vacuoles. A comparison is made between the ultrastructure of the stratified layers in situ and those described by others in differentially ultracentrifuged homogenates.