DIEL OSCILLATIONS IN THE PHOTOSYNTHESIS-IRRADIANCE RELATIONSHIP OF A PLANKTONIC MARINE DIATOM1

The amplitude of diel oscillations in photosynthesis as a function of irradiance varied with the growth phase in a marine phytoplankton species. The common centric diatom (Bacillariophyta), Ditylum brightwellii (West) Grun., showed strong periodicity in the photosynthesis-irradiance (P-I) relationship, which damped progressively from early to late exponential and stationary phase. These findings suggest that short-term temporal characteristics of phytoplankton production depend on factors which affect growth, and that the amplitude is most enhanced at maximal growth rates likely to be encountered in the natural environment.     

FLOW CYTOMETRIC ANALYSIS OF SPERMATOGENESIS IN THE DIATOM THALASSIOSIRA WEISSFLOGII (BACILLARIOPHYCEAE)1

Flow cytometry was used to detect and quantify sexual differentiation in the centric diatom Thalassiosira weissflogii (Grun.). Size (light scatter), chlorophyll, protein and DNA contents were measured for each cell throughout the process of differentiation. Male gametes were small round cells characterized by one complement of DNA and a lower protein and chlorophyll content than vegetative cells. Male gamete formation was induced by a long period of darkness (2 days) followed by a transfer to continuous light. Up to 30% of the initial cell population produced male gametes which appeared in the culture 14 h after release from darkness. Male gamete production was also detected in exponentially growing cultures in continuous light, but to a much smaller degree.     

FLOW CYTOMETRIC ANALYSIS OF SPERMATOGENESIS IN THE DIATOM THALASSIOSIRA WEISSFLOGII (BACILLARIOPHYCEAE)1

Flow cytometry was used to detect and quantify sexual differentiation in the centric diatom Thalassiosira weissflogii (Grun.). Size (light scatter), chlorophyll, protein and DNA contents were measured for each cell throughout the process of differentiation. Male gametes were small round cells characterized by one complement of DNA and a lower protein and chlorophyll content than vegetative cells. Male gamete formation was induced by a long period of darkness (2 days) followed by a transfer to continuous light. Up to 30% of the initial cell population produced male gametes which appeared in the culture 14 h after release from darkness. Male gamete production was also detected in exponentially growing cultures in continuous light, but to a much smaller degree.     

FILAMENT FORMATION IN THE DIATOM MELOSIRA GRANULATA1

The frequency and position of separation valves in filaments of Melosira granulata were observed both in laboratory cultures and in natural habitats. The numerical relationship between mean filament length in a population and the proportion of separation valves produced (the separation valve index-SVI) has also been determined. The data indicate that filament length is controlled through the production of separation valves and that the SVI gives an indication of the mean filament length. Two series of cell divisions (the first producing the separation valves) are necessary before separation of the filament can occur. Paris of separation valves occur predominantly towards the middle of the filament. We suggest that the SVI may prove to be a useful indicator of mean filament length, and hence of ecological change, even when the filaments are no longer intact, as, for example, in sediment core samples and on permanent slides of acid-cleaned collections.     

THE INLAND CHAETOCEROS (BACILLARIOPHYCEAE) SPECIES OF NORTH AMERICA1

Five taxa of Chaetoceros occur in inland waters of North America. These most commonly occur in waters with elevated total dissolved solids in arid regions of the western United States and Canada. Chaetoceros amanita Cleve-Euler is characterized by consistently forming relatively long chains of cells and having very spinose primary resting spore valves. Chaetoceros elmorei Boyer also forms long chains of cells which are connected by evident valvar processes; spores are nearly always smooth. Chaetoceros muelleri Lemm. may form short chains with processes between sibling valves, but also produces solitary cells lacking processes. Chaetoceros muelleri var. subsalsum (Lemm.)Johansen et Rushforth is similar to the nominate but never produces cells with Processes. Both of the C. muelleri varieties produce spores with smooth primary valves. Chaetoceros simplex Ostenfeld is characterized by a noncolonial habit, cells lacking processes and the production of resting spores with warty to some what spinose primary valves.     

INTERACTIVE EFFECTS OF COPPER AND SILICIC ACID ON RESTING SPORE FORMATION AND VIABILITY IN A MARINE DIATOM1

The toxic effects of copper on resting spore formation and viability in the marine diatom Chaetoceros protuberans Lauder were determined both with and without silicic acid added to the medium. With silicic acid available, partial inhibition of resting spore formation occurred only at the highest cupric ion activity (pCu 8.6), while the percentage of cells forming spores at pCu's 10.2 and 11.3 was nearly the same as in the controls. Without silicic acid added to the medium, sporulation was completely inhibited at pCu 8.6 and greatly inhibited, at pCu 10.2. At pCu 11.3 and in the controls, the rate of spore formation was less than 50%. The results indicate that the inhibition of resting spore formation by copper is related to the concentration of silicic acid available to cells of C protuberans. This is consistent with previous studies which show that copper toxicity during vegetative growth involves interference with silicification in diatoms and is a Junction of the silicic acid concentration of the medium. Viable resting spores of C. protuberans were still present in cultures following exposure to elevated copper concentrations during a 100-day incubation period. This indicates that resting spores can serve to enhance diatom survival in areas polluted by heavy metals.     

INTERACTIVE EFFECTS OF COPPER AND SILICIC ACID ON RESTING SPORE FORMATION AND VIABILITY IN A MARINE DIATOM1

The toxic effects of copper on resting spore formation and viability in the marine diatom Chaetoceros protuberans Lauder were determined both with and without silicic acid added to the medium. With silicic acid available, partial inhibition of resting spore formation occurred only at the highest cupric ion activity (pCu 8.6), while the percentage of cells forming spores at pCu's 10.2 and 11.3 was nearly the same as in the controls. Without silicic acid added to the medium, sporulation was completely inhibited at pCu 8.6 and greatly inhibited at pCu 10.2. At pCu 11.3 and in the controls, the rate of spore formation was less than 50%. The results indicate that the inhibition of resting spore formation by copper is related to the concentration of silicic acid available to cells of C. protuberans. This is consistent with previous studies which show that copper toxicity during vegetative growth involves interference with silicification in diatoms and is a function of the silicic acid concentration of the medium. Viable resting spores of C. protuberans were still present in cultures following exposure to elevated copper concentrations during a 100-day incubation period. This indicates that resting spores can serve to enhance diatom survival in areas polluted by heavy metals.     

BLOOMS OF SURF-ZONE DIATOMS ALONG THE COAST OF THE OLYMPIC PENINSULA,WASHINGTON. VI. DAILY PERIODICITY PHENOMENA ASSOCIATED WITH CHAETOCEROS ARMATUM IN ITS NATURAL HABITAT1,2

Microscope observations of chains of Chaetoceros T. armatum T. West, collected, from its natural habitat, at Copalis Beach, Washington, have been made. A regular daily periodicity in the occurrence of cell division stages and length of the chains was found on each occasion (November, February, April, May, August). During the night, short chains of 3 cells predominated. In early morning hours, average chain lengths increased due to cell elongation, and nuclear and protoplast division occurred. This correlated with the time of day when cell masses of C. armatum first appeared at the surface of the water. In the late afternoon, the new silica valves of the daughter protoplasts were formed. Formation of special bristle-bearing end valves effected the separation of long chains into shorter chains composed predominantly of 3 cells. (Cultured material was used to verify some of these stages.) This correlated with the time of day when cell masses disappeared from the water surface and dispersed.     

并基角毛藻若干分类学疑问的初步探讨

并基角毛藻是硅藻门角毛藻属的典型种类, 以“相邻角毛基部并行融合”作为标志性特征, 但该特征是否稳定, 及如何赋予其分类学价值?尚存较多争议。研究以符合现存并基角毛藻分类标准的藻株作为目标藻株, 采用毛细管显微操作技术从我国沿海代表水域分离藻株, 建立了目标藻株的单克隆培养株系, 利用光学显微镜、扫描电镜和透射电镜分别对其形态学特征进行观察, 同时还扩增了核糖体大亚基编码基因的D1-D3区序列, 并构建系统进化树。综合分析形态学特征和分子系统学数据, 初步得到以下结论: (1)具有相同遗传特征的并基角毛藻株系在“相邻角毛基部并行融合”特征上具有不稳定性, 即“相邻角毛基部并行融合”不能作为并基角毛藻种类的标志性特征; (2)并基角毛藻和洛氏角毛藻作为相似种类, 均显示出较高的物种多样性, 高于目前的认知, 预示着或许存在隐形种或拟隐形种; (3)推测并基角毛藻单胞变型只是并基角毛藻原种的一个生活史阶段, 是并基角毛藻的同种异名, 不宜继续保留其独立的分类学地位。     

PHYSIOLOGICAL VARIABILITY WITHIN TEN STRAINS OF CHAETOCEROS MUELLERI (BACILLARIOPHYCEAE)1

The effects of temperature, ionic composition, and conductivity on growth rates of ten strains of Chaetoceros muelleri Lemmerman (mostly var. subsalsum Johan. & Rushf.) were studied. Lipid content of stressed and unstressed cells and fatty acid composition were also determined. Considerable physiological variability was observed in the ten strains, although principal components analysis of physiological data indicated that all strains fell into one of two major groups: C. muelleri (var. muelleri and var. subsalsum) and an undescribed Chaetoceros species morphologically close to C. muelleri var. subsalsum. A high degree of agreement was found among morphological, physiological, and biochemical data sets, indicating that physiological and biochemical data may be helpful in making taxonomic decisions in diatoms, particularly in taxa with few morphological characters. We also conclude that nonmorphological characters such as those employed in the present study can be used to test phylogenetic hypotheses formulated from traditional morphological data.     

DIEL CHANGES IN THE COMPOSITION OF PHOTOSYNTHETIC PIGMENTS AND CELLULAR CARBON AND NITROGEN IN CHATTONELLA ANTIQUA (RAPHIDOPHYCEAE)1

An axenic clonal culture of Chattonella antiqua (Hada) Ono was grown on a 12: 12 h LD cycle in a laboratory culture tank containing 1 m³ of f/2 medium. Diel changes in mean cell volume, cellular carbon (carbon content per cell), C/N ratio, cellular Chl a, Chl a/c ratio and carotenoid composition were observed. Mean cell volume and cellular C, N and pigments increased during the light period as a result of photosynthesis and decreased with increase of cell concentration by phased cell division during the dark period. These changes indicated that carbon assimilation and pigment synthesis occurred together during the light period. However, the patterns of increase were not the same since different diel patterns were also found in the ratios of C/N and chl a/c. Photosynthetic pigments were analyzed by reversed-phase high-performance liquid chromatography with ion-pairing solution. This analysis showed that the dominant carotenoids in C. antiqua were fucoxanthin, violaxanthin and β-carotene. Diel patterns of Chls a and c were similar to that of fucoxanthin but different from those of violaxanthin and β-carotene. The cellular contents of Chl a, fucoxanthin and carbon increased in a parallel manner during the light period. On the other hand, the increase of violaxanthin was restricted to only a few hours at the beginning of the light period during cell division cycles.     

SEASONAL PATTERN OF DIEL PERIODICITY IN PHOTOSYNTHESIS BY POLAR PHYTOPLANKTON: SPECIES-SPECIFIC RESPONSES1

The diel patterns of light-saturated and light-limited photosynthesis were measured for three diatom species in McMurdo Sound, Antarctica during the transition from late austral winter to summer. Maximum photosynthetic capacity occurred around mid-day during September, when there was a well defined light/dark cycle, and progressively shifted to about midnight by late october when irradiance was continuous. There was a concomitant shift in minimum photosynthetic capacity from midnight to midday. Rates of light-saturated and -limited photosynthesis covaried, and the magnitude of seasonal and diel changes in photosynthetic characteristics were similar. The linear relationship between light-saturated and -limited photosynthesis suggests that the shapes of the photosynthesis-irradiance curves remained relatively constant over the day and througout the season. The unique diel patterns of photosynthesis of these polar phytoplankton appear to be a response to the persistently low, yet continuous irradiance of the polar summer.     

OBSERVATIONS ON THE SURVIVAL AND GERMINATION OF RESTING SPORES OF THREE CHAETOCEROS (BACILLARIOPHYCEAE) SPECIES1,2

Resting spores (hypnospores) of Chaetoceros diadema (Ehrenberg) Gran, Chaetoceros vanheurckii Gran, and Chaetoceros didymus Ehrenberg were collected from a large plastic enclosure moored in Saanich Inlet, B.C., Canada. The effects of combinations of temperature and irradiance on the germination of these resting spores were investigated. Nutrient uptake, carbon fixation, and changes in the photosynthetic capacity of the germinating spores were also examined. Resting spores germinated optimally at combinations of temperature and irradiance similar to those in the environment during sporulation. They did not germinate at irradiances 1.3 μEin m^?2 s^?1 or temperatures >25.3° C. Nitrate, phosphate and silicate were taken up after the resting spores had germinated and resumed vegetative growth. Chlorophyll a fluorescence in vivo, and the DCMU-induced increase in in vivo fluorescence also increased after the resting spores had germinated. Resting spores began to fix carbon as soon as they were placed in light. Spores remained viable for at least 645 d. The length of time between first exposure to light and germination did not change during this period; however, the percentage of viable resting spores decreased markedly. None of the Chaetoceros spores germinated after 737 d of storage at 2–4° C in darkness.     

CELL DIVISION AND MORPHOGENESIS OF THE CENTRIC DIATOM CHAETOCEROS DECIPIENS (BACILLARIOPHYCEAE) I. LIVING CELLS

Like other diatoms, living cells of Chaetoceros decipiens Cleve expand lengthwise before they divide. During prophase, the nucleolus disappears in about 30 s. The spindle is very small but anaphase chromosome separation can be seen. Following rapid cleavage, the protoplasts contract, plasmolyzing slightly and transforming the cleavage furrow into a lens-shaped opening between daughter cells. During valve initiation, the surface of the furrow is molded slightly into the shape of the mature valve face. Then daughter cells expand further, becoming fully turgid as they open the slots in the girdle bands through which the setae will grow. Soon, delicate protrusions push through the girdle bands and develop into the setae, which are very sensitive: any disturbance will immediately stop their steady growth. Healthy setae display soft, mobile tips and tiny organelles (mitochondria) actively move along the lumen. Their curvature and uniform diameter is controlled during growth with exquisite precision, and in optimal conditions, they can become very long. At their initiation, cells appear fully turgid; however, many cells soon become slightly plasmolyzed during seta growth. This observation strongly suggests that turgor pressure cannot be responsible for driving extension; the possible mechanism is discussed in the following paper.     

VALVE MORPHOGENESIS AND THE MICROTUBULE CENTER IN THREE SPECIES OF THE DIATOM NITZSCHIA1

The relationship of cell organelles to valve morphogenesis was investigated in three species of Nitzschia. One, N. sigmoidea (Nitzsch) W. Sm., showed consistent ability to generate both nitzschioid and hantzschioid symmetry in daughter cells following cytokinesis; the other two maintained nitzschioid symmetry stably. From previous work with Hantzschia, a certain sequence of events could be anticipated in the cytoplasm. In two significant areas–the behavior of the Microtubule Center (MC) and its microtubule (MT) system, and the central origin of the silicalemma–not only were the results unexpected, but the three species showed fundamental differences among themselves. In N. sigmoidea, the silicalemma (and the future raphe region) arises centrally on the cleavage furrow, and after some lateral expansion, the silicalemmas and their associated organelles move in opposite directions in daughter cells, so that the raphe and the raphe canals end up along the girdle side of the cell as expected. However, the MCs never become associated with their silicalemma, remaining throughout near the girdle bands. In N. sigma (Kütz) W. Sm., the silicalemmas arise centrally and after lateral growth, move in opposite directions to generate nitzschioid symmetry. In this case, the MCs move to the vicinity of but never close to the silicalemmas, and follow them distantly during their lateral movement. In N. tryblionella Hantzsch, the new silicalemmas arise opposite one another, on one side of the daughter cells; each MC soon moves very close to its silicalemma, and remains thus through most of valve morphogenesis. Later, only one silicalemma/MC complex moves laterally, establishing the nitzschioid symmetry in both daughter cells. In all three species, as in Hantzschia, linear arrays of mitochondria aligned along MTs occupy the forming raphe canal, and microfilaments line the outer edge of the expanding silicalemma. The fibulae (the wall struts arching across the raphe canal) in Hantzschia always grow from the valve surface to the girdle surface of the forming valves. In these three Nitzschiae, this invariably happens in only one daughter cell of any pair; in the other, all the fibulae grow from the girdle surface to the valve surface. An explanation of these variations is proposed: that the morphogenetic machinery of Nitzschia and Hantzschia have a common origin, with present Nitzschiae having undergone considerable diversification at the intracellular level, causing the unstable cell symmetry exhibited by several modern species. Perhaps a taxonomic distinction between Hantzschia and Nitzschia lies in whether the morphogenetic machinery associated with valve morphogenesis moves laterally in the same or in opposite directions.     

ROLE OF LIGHT AND THE CELL CYCLE ON THE INDUCTION OF SPERMATOGENESIS IN A CENTRIC DIATOM1

The centric diatom, Thalassiosira weissflogii Grun., can be induced to undergo spermatogenesis by exposing cells maintained at saturating levels of continuous light to either dim light or darkness. Using flow cytometry to determine the relative DNA and chlorophyll content per cell, the number of cells within a population that responded to and induction signal was measured. From 0 to over 90% of a population differentiated into male gametes depending upon both the induction trigger and the population examined, regardless of the average cell size of the population. Through the use of synchromized cultures, we demonstrated that responsiveness to an induction trigger was a function of cell cycle stage; cells in early G₁ were not yet committed to complete mitosis and were induced to form male gametes, whereas cells further along in their cell cycle were unresponsive to these same cues. A simple model combining the influence of light on the mitotic cell cycle and on the induction of spermatogenesis is proposed to explain the observed diversity in population responses to changes in light conditions.     

DIATOM MINERALIZATION OF SILICIC ACID IV. KINETICS OF SOLUBLE Si POOL FORMATION IN EXPONENTIALLY GROWING AND SYNCHRONIZED NAVICULA PELLICULOSA1

Silicic acid taken up from the growth medium by Navicula pelliculosa (Bréb.) Hilse was shown to enter at least two compartments: i) soluble pools; ii) insoluble fraction comprised predominantly of the silica frustule. Soluble Si pools were extracted by a variety of agents from cells uniformly labeled for ten generations in medium containing ⁶⁸Ge-Si(OH)₄. 100 C water soluble and 0 C perchloric acid (PCA) soluble Si pools of 680 mM Si·l^?1 and 490 mM Si·l^?1 cell water represented 13 and 9%, respectively, of total, cell Si in exponential growth phase cells. Uniformly labeled cells synchronized by the combined synchronization technique accumulate at the cell cycle stage where silica frustule development is initiated. These cells contain water and PCA soluble pools of 10 nmol Si·10⁶ cells^?1 and-8.8 nmol Si·10⁶ cells^?1, respectively. On addition of Si(OH)₄, a rapid uptake ensues allowing the Si pool to expand 2.5-fold, apparently to provide precursors of the silica frustule.     

TAXONOMY AND FRUSTULAR STRUCTURE OF THE MARINE CENTRIC DIATOM PARALIA SULCATA1

Light and electron microscopy were used to investigate the complex structure of the frustule of Paralia sulcata (Ehrenb.) Cleve. Rimoportulae are reported for the first time in this diatom and two types of linking processes are described. The ease with which the cingulum is lost is explained with regard to its attachment to the valve. Two kinds of heterovalvy were observed and the taxonomic significance of one of these is discussed. The validity of Heiberg's genus Paralia is confirmed and a type slide of the species is designated.