Resting spore formation ofLeptocylindrus danicus (Bacillariophyceae) during short time-scale upwelling and its significance as predicted by a simple model

Resting spore formation during short time-scale upwelling and its significance were investigated in the field and by a simple theoretical model. Field observations of spore formation ofLeptocylindrus danicus were made off Izu Peninsula, Japan. A rapid increase in ratio of resting spore to vegetative cell numbers indicated thatL. danicus formed resting spores quickly as a response to nutrient depletion in the upwelled water, although only a very low number of resting spores was found in the upwelling. A simple model was constructed to investigate the possible advantages of spore formation during short time-scale upwelling. This showed that there is a critical time-scale for resting spore formation to be advantageous. The nutrient depletion period of the upwelling off Izu was shorter than the critical time-scale determined by the model. Rapid-sinking of resting spores may increase further the critical time-scale, unless spores return with upwelling water. For short time-scale upwelling, the vegetative cell may be better suited than the resting spore for enduring a short period of nutrient depletion. Contribution from Shimoda Marine Research Center, University of Tsukuba, No. 475.     

氮、硅限制对赤潮藻扁面角毛藻休眠孢子形成的影响

休眠孢子的形成对于赤潮藻种群的保存、延续以及分布扩散等均具有重要的意义。通过单因子营养限制研究氮、硅对赤潮藻扁面角毛藻（Chaetoceros compressus）休眠孢子形成的影响,结果表明：培养基中氮的初始浓度对休眠孢子的出现时间有一定影响。氮的初始浓度越低,休眠孢子出现的时间越早：反之,氮的初始浓度越高,休眠孢子出现的时间越晚。氮缺乏是硅藻形成休眠孢子的必需条件之一,当培养基中氮含量低于10μmol·L^-1时,扁面角毛藻可以形成休眠孢子。氮缺乏诱发的休眠孢子的形成需要大量的硅,当培养基中硅含量低于23μmol·L^-1时,即使氮缺乏,扁面角毛藻也几乎不再继续形成休眠孢子。这说明硅藻休眠孢子的形成不仅受氮浓度的影响,还与硅浓度有关。     

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.     

氮、硅限制对赤潮藻扁面角毛藻休眠孢子形成的影响

休眠孢子的形成对于赤潮藻种群的保存、延续以及分布扩散等均具有重要的意义。通过单因子营养限制研究氮、硅对赤潮藻扁面角毛藻 (Chaetoceros compressus )休眠孢子形成的影响, 结果表明: 培养基中氮的初始浓度对休眠孢子的出现时间有一定影响。氮的初始浓度越低, 休眠孢子出现的时间越早; 反之, 氮的初始浓度越高, 休眠孢子出现的时间越晚。氮缺乏是硅藻形成休眠孢子的必需条件之一, 当培养基中氮含量低于10 mmol.L^-1时, 扁面角毛藻可以形成休眠孢子。氮缺乏诱发的休眠孢子的形成需要大量的硅, 当培养基中硅含量低于23 mmol.L^-1时, 即使氮缺乏, 扁面角毛藻也几乎不再继续形成休眠孢子。这说明硅藻休眠孢子的形成不仅受氮浓度的影响, 还与硅浓度有关。     

STUDIES ON MARINE PLANKTON DIATOMS. II. RESTING SPORE MORPHOLOGY1

A morphological study of resting spores in five marine planktonic diatom species using electron microscopy indicates that Bacteriastrum delicatulum Cleve and Leptocylindrus danicus Cleve spores bear little resemblance, to their vegetative cells. Detonula confervacea (Cleve) Gran and Stephanopyxis turris (Grev. & Arn.) Ralfs spores have several features in common with their vegetative cells, and Rhizosolenia setigera Brightwell lies between the two extremes. The function of resting spores in relation to diatom life cycles is briefly discussed. Spore formation may be a primitive characteristic in the life cycle and may no longer have significant survival value for the species.     

“INTERNAL THECAE” OF EUNOTIA SOLEIROLII (BACILLARIOPHYCEAE): DEVELOPMENT,STRUCTURE AND FUNCTION AS RESTING SPORES1

The “double thecae” or “internal septa” of Eunotia soleirolii (Kütz.) Rabenh, are shown to represent the thecae of resting spores, as characterized by their physiology, as well as morphology. They differ from all resting spores of centric diatoms by the formation of both their valves as a result of unequal cell divisions; and, from the majority of centric spores by the presence of several girdle bands in both their thecae. Spore formation can be induced by high or low pH, high temperature (24 C), and iron, silica, phosphate or nitrate deficiencies, whereas low temperatures defer it. Spores do not germinate directly, but dormancy can be removed by dark treatments (–2 to 15 C) for a minimum of 4–5 wk. Longer dark treatments result in higher germination rates. At 15 C, a minimum of 2 mo is required and 4 mo is better. Heat treatments (27–42 C) are ineffective, but may shorten the dormancy-breaking subsequent cold period. Instances of secondary dormancy, as well as relative dormancy, were observed. Germination usually occurs in the light between 2 and 21 C. An equal division of the spore is followed by unequal divisions of both new cells with only the two resulting large cells being viable. The experiences in the laboratory aided the discovery of stages of spore germination in nature.     

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.     

Population dynamics of an ice-associated diatom, Thalassiosira australis Peragallo, under fast ice near Syowa Station, East Antarctica, during austral summer

A clear shift from vegetative cells to auxospores and resting spores in Thalassiosira australis was observed in the water column and sinking fluxes under the fast ice near Syowa Station in the austral summer of 2005/2006. This is the first report of the auxosporulation by T. australis in situ. Resting spores were also observed in the sediment even before new spore formation, suggesting that T. australis can overwinter in the sediment. Heterotrophic dinoflagellates ingested and digested vegetative cells and auxospores but did not digest resting spores, suggesting a high tolerance of resting spores to grazing by heterotrophic dinoflagellates. We discuss the possible life history and overwintering strategy that T. australis uses in an Antarctic coastal area to cope with the unpredictable timing of sea ice growth and decay.     

Dietary and Feeding Differences Between Sympatric Propithecus diadema diadema and Indri indri

We analyzed the dietary profiles and feeding behaviors of Propithecus diadema diadema and Indri indri in a community of animals that reside in midaltitude rain forest within Madagascar's Mantadia National Park. Propithecus diadema diadema ate a diverse mixture of fruits, seeds, flowers and young leaves, while the bulk of the diet of Indri indri consisted of young leaves, which resulted in significantly higher levels of fat and water-soluble carbohydrates in foods eaten by Propithecus diadema diadema. Fiber values of items eaten are high (54% NDF) for both species, though not significantly different between them. The preference for immature foliage by Indri indri suggests that their overall intake of fiber is greater than that of Propithecus diadema diadema, which had a high proportion of non-leaf material in their diet. We propose that differences in gut morphology between the two indriids contribute to their disparate diets. Levels of secondary compounds were high in certain food items, but overall they are also not significantly different between the two indriid diets. Propithecus diadema diadema exhibited a strong preference for 2 alkaloid-containing seed species, while no food of Indri indri contained alkaloids. In addition, Propithecus diadema diadema consumed a higher diversity of plant species on a daily/yearly basis, exhibited more feeding bouts on a daily basis, and their feeding bouts (on young leaves, fruit and flowers) are significantly shorter in duration than those of Indri indri. Furthermore, Propithecus diadema diadema had twice the number of geophagic episodes of Indri indri.     

Synchytrium solstitiale: reclassification based on the function and role of resting spores

Studies were made about resting spores of Synchytrium solstitiale, a chytrid that causes false rust disease of yellow starthistle (YST). During evaluation of this fungus for biological control of YST, a protocol for resting spore germination was developed. Details of resting spore germination and study of long-term survival of the fungus were documented. Resting spores from dried leaves germinated after incubating them on water agar at least 7 d at 10-15 C. Resting spores were viable after storage in air-dried leaves more than 2 y at room temperature, suggesting they have a role in off-season and long-term survival of the fungus. Each resting spore produced a single sorus that contained a single sporangium, which on germination released zoospores through a pore. YST inoculated with germinated resting spores developed symptoms typical of false rust disease. All spore forms of S. solstitiale have been found to be functional, and the life cycle of S. solstitiale has been completed under controlled laboratory and greenhouse conditions. Resting spore galls differ from sporangial galls both morphologically and biologically, and in comparison, each sporangial gall cleaves into several sori and each sorus produces 5-25 sporangia that rupture during release of zoospores. For this reason S. solstitiale should be reclassified as diheterogallic sensu Karling (Am J Bot 42:540-545). Because resting spores function as prosori and produce an external sorus, S. solstitiale is best placed in into the subgenus Exosynchytrium.     

Thalassiosira antarctica (Bacillariophyceae): Vegetative and resting stage ultrastructure of an ice-related marine diatom

Summary The ultrastructure of T. antarctica var. antarctica vegetative and resting stages are compared using light and transmission electron microscopy. Resting spores contain noticeably more lipid reserves than do vegetative cells. Numerous mitochondria and generally fewer numbers of other organelles are eliminated from spores into an abortive daughter cell when the spore formation division sequence is terminated. The remaining spore contents are a compact arrangement of organelles with lipid bodies predominating. These two stages are thus ultrastructurally distinct, and differences in their chemical composition can be manifested as cytological modifications.     

The role of temperature on in vivo resting spore formation of the aphid-specific pathogen Pandora nouryi (Zygomycota: Entomophthorales) under winter field conditions

Resting spore formation of Pandora nouryi within the host Myzus persicae was investigated in an open cabbage field during two winters: 2007–2008 and 2010–2011. Multivariate correlation analysis of field data revealed that weekly mean temperature was the main abiotic factor governing the probability of resting spore formation in vivo, followed by host density and mean day length, while the effect of season was insignificant. Moreover, the temperature effect on the proportions of resting spore producing cadavers in those aphids infected with P. nouryi was well fit by a logistic equation, and high proportions (0.37–0.95) were found on most of weekly sampling occasions with mean temperatures of less than 10°C. Thus, these resultant analyses support the hypothesis that temperature is the most important abiotic factor controlling P. nouryi resting spore formation in M. persicae under field situations.     

POPULATION DYNAMICS OF THE SPORE-FORMING DIATOM LEPTOCYLINDRUS DANICUS IN NARRAGANSETT BAY. RHODE ISLAND1

A field study to assess resting spore importance in long-and short-term population dynamics of Leptocylindrus danicus ( Ehr.) Gran was carried out, based on the observation that germinating spores produce only vegelative cells of maximum diameter. From May through November 1978 . L. danicus was present in lower Narragansett Bay, but resting spores were not found. May and June bioassays suggest that Bay water could not support sexuality (hence spore formation) in L. danicus during this period. Mean valve diameter decreased abruptly in early June. as did cell number. When mean valve diameter increased abruptly in August, with no resting spores seen, 70 additional clonal isolates showed that one clone had the "normal" sexual and spore-forming life cycle while 69 clones had an "alternate" life cycle lacking sperm and spore formation. Valvar fine structure of the two types differed with respect to the presence or absence of a subcentral valvar pore, and a new subspecific taxon is here distinguished: variety apora. Examination of the early June data indicates the replacement of one population by another. Populations of L. danicus may be sporadically introduced into Narragansett Bay from Gulf Stream rings .     

Resting spore formation in the antarctic diatoms Coscinodiscus furcatus Karsten and Thalassiosira australis Peragallo

Summary The resting spore morphology of the antarctic diatoms Coscinodiscus furcatus Karsten and Thalassiosira australis Peragallo are described. Both species form endogenous resting spores. The spore valve of C. furcatus differ from those of the vegetative cells primarily by (i) a greater convexity and (ii) a coarser and more distinctly fasciculated areolation. This resting spore is identical to the diatom traditionally identified as C. stellaris var. symbolophorus (Grunow) Jørgensen in the Antarctic. The resting spore of T. australis differs from the vegetative cells by (i) a lack of clusters of strutted processes in a modified ring on valve face, (ii) a coarser areolation and tangential rows of areolae and (iii) a narrower and more simply structured girdle. The resting spore valve of T. australis has been described as belonging to a separate species, Actinocyclus excentricus Peragallo.     

LIFE CYLE AND SEXUALITY OF THE FRESHWATER RAPHIDOPHYTE GONYOSTOMUM SEMEN (RAPHIDOPHYCEAE)1

Previously unknown aspects in the life cycle of the freshwater flagellate Gonyostomum semen (Ehrenb.) (Raphidophyceae) are described here. This species forms intense blooms in many northern temperate lakes, and has increased in abundance and frequency in northern Europe during the past decades. The proposed life cycle is based on observations of life cycle stages and transitions in cultures. Viable stages of the life cycle were individually isolated and monitored by time‐lapse photography. The most common processes undertaken by the isolated cells were: division, fusion followed by division, asexual cyst formation, and sexual cyst formation. Motile cells divided by two different processes. One lasted between 6 and 24 h and formed two cells with vegetative cell size and with or without the same shape. The second division process lasted between 10 and 20 min and formed two identical cells, half the size of the mother cell. Planozygotes formed by the fusion of hologametes subsequently underwent division into two cells. Asexual cyst‐like stages were spherical, devoid of a thick wall and red spot, and germinated in 24–48 h. Heterogamete pairs were isogamous, and formed an angle of 0–90° between each other. Planozygote and sexual cyst formation were identified within strains established from one vegetative cell. The identity of these strains, which was studied by an amplified fragment length polymorphism analysis, was correlated with the viability of the planozygote. Resting cyst germination was described using cysts collected in the field. The size and morphology of these cysts were comparable with those formed sexually in culture. The excystment rate was higher at 24°C than at 19 or 16°C, although the cell liberated during germination (germling) was only viable at 16°C. The placement of G. semen within the Raphidophyceae family was confirmed by sequence analysis of a segment of the 18S ribosomal DNA.     

Chaetoceros resting spores in the Gerlache Strait, Antarctic Peninsula

The formation of resting spores in diatoms is a common phenomenon in neritic environments. Here we report on resting spores of the genus Chaetoceros associated with a layer of increased chlorophyll fluorescence, at a depth of more than 200 m, north of Brabant Island and in Wilhelmina Bay, southeast coast of the Gerlache Strait (64°41.0′S, 62°0.5′W). Six species of Chaetoceros were identified by the morphology and size of the resting spores. Given that Chaetoceros spp., both in vegetative cells and as resting spores, are commonly found in Antarctic coastal surface waters, their location at depth could represent the pelagic “waiting” or “seeding” populations mentioned for other environments. Received: 25 November 1996 / Accepted: 16 November 1997     

Monterey Bay Phytoplankton. II. Resting Spore Cycles in Coastal Diatom Populations

The purpose of this study is to examine the role of "restingspores" in natural population cycles of temperature, neriticdiatoms. Resting spores are a conspicuous element of centricdiatom populations in most neritic and some deep-water environmentsof the world oceans. Until recently the spores have been accepted,almost axiomatically, as surviving as a seasonal benthic stagewhich reinoculate the water column at the onset of favorablegrowth conditions. Data to support this hypothesis, however,is limited and contradictory. In my study, I am examining theoccurrence of resting spores in relation to vegetative cellcycles and oceanographic conditions in Monterey Bay, California,a nearshore upwelling-dominated system. I have utilized samplesfrom the water column, from settling-tubes, and from sedimentcores to document the cycles. Resting spore cycles were observed for many neritic species,and spore cycles for abundant species were considered in detail.Resting spore formation was associated with low nitrate concentrationsin surface waters. After spore formation in the water columnspores were found in settling-tubes and in sediment samples.Spore formation was a frequent event and spores were often presentin the water column and in the sediments. Observations of spore cycles in the present study were consistentwith the prevailing idea that resting spores function as benthicresting stages in coastal populations. The pelagic retentionand/or dispersal in coastal circulation cells, however, alsoappeared to be highly likely. The study has demonstrated thepotential for resting spores to function as survival stagesin Monterey Bay and in other regions with similar hydrographicconditions. The present evidence regarding the role of restingspores suggests that resting spores may have a wide range offunctions, and a more inclusive (or alternate) resting sporehypothesis is suggested.     

Internal valves in populations of Meridion circulare from the A’er Mountain region of northeastern China: Implications for taxonomy and systematics

Abstract A population of Meridion circulare var. circulare (Greville) C.A. Agardh from Inner Mongolia was found to produce Innenschalen or internal spores. Examination of this population with light and scanning electron microscopy showed morphological differentiation between vegetative and spore morphologies. Vegetative valves typically bear costae and one rimoportula at the headpole. Spores lack costae and have two rimoportulae, one at the headpole and the other at the footpole. There is plasticity in the production of valve morphologies, and a variety of vegetative valve and spore combinations are evident. This population of M. circulare var. circulare has initial valves of over 90 μm in length, and all of the initial cells encountered are acostate and bear two rimoportulae. These observations suggest that either spores are the product of sexual reproduction, or that initial valves may be produced parthenogenetically in Meridion. Spores as products of the sexual process have not been reported in diatoms previously, and parthenogenesis in Meridion was reported previously but discounted in other published reports. The plasticity of valve morphologies expressed in M. circulare var. ciculare, between vegetative valves and spores (and back) across a short temporal period suggests that diatoms can alter their cell wall structure dramatically and quickly in response to external variables.     

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.