The explosive diversification of the diatom genus Chaetoceros across the Eocene/Oligocene and Oligocene/Miocene boundaries in the Norwegian Sea

Eocene to middle Miocene stratigraphic changes in species richness, abundance and valve size of Chaetoceros resting spores in the Norwegian Sea (DSDP Site 338) were investigated in order to understand past productivity and paleoenvironmental changes in upwelling regions. As a result, drastic resting spore events were recognized in a 6 myr interval across the Eocene/Oligocene boundary (EO Event), the Oligocene/Miocene boundary (OM Event) and in the early middle Miocene (emM Event). The EO Event was characterized by explosive diversification at both the morpho-generic and specific levels, an increase in abundance, and a decrease in valve size from the upper Eocene through the lowest Oligocene. The OM Event was defined by a two-fold increase in species richness. During the emM Event spore abundance decreased rapidly, and species richness and valve size decreased gradually. These changes may indicate changes in the nutrient supply, especially in upwelling regions. The increased species richness suggests a change from a stable water column with a constant nutrient supply in the Eocene to an unstable one with a sporadic nutrient supply by increased vertical mixing in the Oligocene, based on evaluation of the ecologic differences between dinoflagellate cysts and Chaetoceros resting spores. The role of main primary producer might have switched from dinoflagellates and/or nannoplankton in the Eocene to diatoms, especially Chaetoceros, in the Oligocene in the Norwegian Sea. Increased resting spore species richness during the OM Event may show that environmental changes such as global cooling and nutrient mixing led to a diversification of the spore producing genus Chaetoceros. The emM Event might have been affected by changes in paleoceanographic conditions, perhaps a decrease in nutrient supply. This study presents the first paleoceanographic analysis using not only the total resting spore abundance but also the abundances of individual species, and establishes the value of spore taxonomy and diatom analysis including spores.     

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.     

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     

Honey-coloured,sessile Endogone spores: II. Changes in fine structure during spore development

Summary The fine structure of honey-coloured, sessile Endogone spores is described from initiation of the mother spore to dormancy of the resting spore. Three unusual organelles occur viz. pigment granules, large crystals and selfduplicating bacteria-like organisms. The first two are very numerous, and are specifically associated with spore formation. The pigment granules are involved in the deposition of the honey-coloured wall, and change into myelin-like figures when cytoplasm moves from the mother into the resting spore. The crystals, whose function is not known, are most conspicuous just before the resting spore reaches dormancy. The bacteria-like organisms, which may be actinomycete spores living symbiotically in the fungus, multiphy greatly as the spore enters dormancy. The dormant spore contains very little cytoplasm compressed into a fine network between very large polygonal oil globules and large round bodies thought to contain a storage polysaccharide.     

SPORE FORMATION IN THE LIFE CYCLES OF THE DIATOMS CHAETOCEROS DIADEMA AND LEPTOCYLINDRUS DANICUS1

A nitrogen limitation technique elicited the entire life cycle of the marine centric diatoms Chaetoceros diadema (Ehr.) Gran and Leptocylindrus danicus Cleve. In C. diadema the sexual cycle followed the same pattern as in the previously investigated C. didymus. Sexuality took place in narrow diameter cells, only at 2 and 5° C, and was seldom seen. Resting spore formation took place in cells of all sizes and at all temperatures at which the species grew vegetatively (2–15° C). The L. danicus life cycle is probably unique among diatoms. Nitrogen depletion induced sexuality in the entire culture at 10 and 15° C if the cell diameter was narrow (3–8 μm). Auxospore formation was followed by resting spore formation directly within the auxospore. In C. diadema, as in most centric diatoms, resting spores are not an obligate part of the life cycle, but they are in L. danicus. Resting spore formation is a versatile adaptive response in C. diadema, depending only on nitrogen depletion, although promoted by low temperatures. In L. danicus the linkage to the sexual process sharply limits conditions under which resting spores can form.     

CotG controls spore surface formation in response to the temperature of growth in Bacillus subtilis

Bacterial spores of the Bacillus genus are ubiquitous in nature and are commonly isolated from a variety of diverse environments. Such wide distribution mainly reflects the spore resistance properties but some Bacillus species can grow/sporulate in at least some of the environments where they have been originally isolated. Growing and sporulating at different conditions is known to affect the structure and the resistance properties of the produced spore. In B. subtilis the temperature of growth and sporulation has been shown to influence the structure of the spore surface throughout the action of a sporulation-specific and heat-labile kinase CotH. Here we report that CotG, an abundant component of the B. subtilis spore surface and a substrate of the CotH kinase, assembles around the forming spore but also accumulates in the mother cell cytoplasm where it forms aggregates with at least two other coat components. Our data suggest that the thermo-regulator CotH contributes to the switch between the coat of 25°C and that of 42°C spores by controlling the phosphorylation levels of CotG that, in turn, regulates the assembly of at least two other coat components.     

Diversity in the section Compressa of the genus Chaetoceros (Bacillariophyceae), with description of two new species from Chinese warm waters

Monoclonal strains of Chaetoceros species were collected from Chinese warm waters. Vegetative cells and resting spores were examined using light microscopy as well as scanning and transmission electron microscopy. Combined morphological and DNA sequence data from the hypervariable D1–D3 region of the nuclear ribosomal large subunit showed the presence of two new species within the section Compressa, herein described as Chaetoceros bifurcatus sp. nov. and C. millipedarius sp. nov. Both species possessed features typical of the section Compressa, but C. bifurcatus was characterized by the heavy intercalary setae fusing and extending together for a distance before diverging. The heavy setae were not visually contorted, lacking poroids and spines, in contrast with how the section Compressa has been defined. Chaetoceros millipedarius was characterized by a horizontal stagger of ordinary intercalary setae when the chain was seen in broad girdle view, making the chain look like a millipede. The two new species were supported by molecular phylogenetic analyses with C. bifurcatus sister to C. contortus var. ornatus, while C. millipedarius was sister to C. contortus var. contortus. Based on the morphological features exhibited in C. bifurcatus, the diagnosis of the section Compressa was emended to exclude “contorted” heavy setae and compressed valve faces.     

Mutations specific to spore maturation in the asexual fruiting body of dicytostelium discoideum

Three mutations affecting spore maturation in the asexual fruiting body of Dictyostelium discoideum are assigned to a new locus, sprJ, on linkage group IV. Strains carrying mutations at the sprJ locus do not form mature spores, yet the cell patterning (spore, stalk and disc cell ratios) is apparently normal. These mutations will be useful to delineate branch points between the cell patterning and spore maturation pathways. There are some unusual features of the sprJ-containing mutants. In particular each of the parent strains of the three mutants has incomplete spore maturation as determined by colony-forming ability after heat shocking at 45°C. A mutation allowing growth in the presence of benlate (600 μg/ml), benA351, is mapped to linkage group I.     

Resting Spore Dormancy and Infectivity Characteristics of the Potato Powdery Scab Pathogen Spongospora subterranea

The soil‐borne potato pathogen Spongospora subterranea persists in soil as sporosori, which are aggregates of resting spores. Resting spores may germinate in the presence of plant or environmental stimuli, but direct evidence for resting spore dormancy is limited. A soilless tomato bait plant bioassay and microscopic examination were used to examine features of S. subterranea resting spore dormancy and infectivity. Dried sporosori inocula prepared from tuber lesions and root galls were infective after both short‐ and long‐term storage (1 week to 5 years for tuber lesions and 1 week to 1 year for root galls) with both young and mature root galls inocula showing infectivity. This demonstrated that a proportion of all S. subterranea resting spores regardless of maturity exhibit characteristics of stimuli‐responsive dormancy, germinating under the stimulatory conditions of the bait host plant bioassay. However, evidence for constitutive dormancy within the resting spore population was also provided as incubation of sporosorus inoculum in a germination‐stimulating environment did not fully exhaust germination potential even after 2.4 years. We conclude that S. subterranea sporosori contain both exogenous (stimuli‐responsive) and constitutively dormant resting spores, which enables successful host infection by germination in response to plant stimuli and long‐term persistence in the soil.     

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.     

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.     

Variation of upwelling/oceanic conditions during the latest Pleistocene through Holocene off the central Peruvian coast: A diatom record

Diatoms are well preserved in partly laminated Holocene and late Pleistocene sediments on the upper continental slope of central Peru. Accumulation rates of diatom species previously recognized as tropical (oceanic, warm water) and subtropical (neritic, cooler water, upwelling) changed markedly downcore in Kasten cores obtained at 11°15′S, 12°58′S, and 13°37′S latitude. Subtropical species (e.g., Thalassiosira eccentrica “group”, Thalassiothrix frauenfeldii, Chaetoceros spp. resting spores, Delphineis spp.) predominated in late Pleistocene sediments of the two southern cores. A mix of tropical and subtropical diatom assemblages occurred in latest Pleistocene sediments, while tropical species (e.g., Coscinodiscus nodulifer/radiatus, Pseudoeunotia doliolus, Thalassionema nitzschioides var. parva) predominated in the late Holocene sediments. In the northern core latest Pleistocene sediments also contained a mix of tropical and subtropical species. Similarly, late Holocene sediments contained predominately tropical species of diatoms. Neoglacial periods (200–400 yr B.P., 2000–2700 yr B.P.), however, were sometimes characterized by a greater abundance of subtropical species and punctuated periodically with blooms of Skeletonema costatum and Rhizosolenia shrubsolei. It is proposed that during times of Southern Hemisphere cooling, Peruvian coastal waters north of 12°S were subjected to episodic mixing with warmer water masses before being overwhelmed by tropical waters that may have either passed southward across a weakened Equatorial Front or arose from intensified offshore countercurrents. Coastal waters south of 12°S latitude were probably unaffected by the tropical influence. Rather, these waters may have undergone intensified upwelling during the last Glacial epoch.     

Test method development to evaluate hot,humid air decontamination of materials contaminated with Bacillus anthracis ∆Sterne and B. thuringiensis Al Hakam spores

Aims

To develop test methods and evaluate the survival of Bacillus anthracis ?Sterne and Bacillus thuringiensis Al Hakam spores after exposure to hot, humid air.

Methods and Results

Spores (>7 logs) of both strains were dried on six different test materials. Response surface methodology was employed to identify the limits of spore survival at optimal test combinations of temperature (60, 68, 77°C), relative humidity (60, 75, 90%) and time (1, 4, 7 days). No spores survived the harshest test run (77°C, 90% r.h., 7 days), while > 6·5 logs of spores survived the mildest test run (60°C, 60% r.h., 1 day). Spores of both strains inoculated on nylon webbing and polypropylene had greater survival rates at 68°C, 75% r.h., 4 days than spores on other materials. Electron microscopy showed no obvious physical damage to spores using hot, humid air, which contrasted with pH‐adjusted bleach decontamination.

Conclusions

Test methods were developed to show that hot, humid air effectively inactivates B. anthracis ?Sterne and B. thuringiensis Al Hakam spores with similar kinetics.

Significance and Impact of the Study

Hot, humid air is a potential alternative to conventional chemical decontamination.     

Two new species in the Chaetoceros socialis complex (Bacillariophyta): C. sporotruncatus and C. dichatoensis,and characterization of its relatives,C. radicans and C. cinctus

The diatom genus Chaetoceros is one of the most abundant and diverse phytoplankton in marine and brackish waters worldwide. Within this genus, Chaetoceros socialis has been cited as one of the most common species. However, recent studies from different geographic areas have shown the presence of pseudo‐cryptic diversity within the C. socialis complex. Members of this complex are characterized by curved chains (primary colonies) aggregating into globular clusters, where one of the four setae of each cell curves toward the center of the cluster and the other three orient outwards. New light and electron microscopy observations as well as molecular data on marine planktonic diatoms from the coastal waters off Chile revealed the presence of two new species, Chaetoceros sporotruncatus sp. nov. and C. dichatoensis. sp. nov. belonging to the C. socialis complex. The two new species are similar to other members of the complex (i.e., C. socialis and C. gelidus) in the primary and secondary structure of the colony, the orientation pattern of the setae, and the valve ultrastructure. The only morphological characters that can be used to differentiate the species of this complex are aspects related to resting spore morphology. The two newly described species are closely related to each other and form a sister clade to C. gelidus in molecular phylogenies. We also provide a phylogenetic status along with the morphological characterization of C. radicans and C. cintus, which are genetically related to the C. socialis complex.     

Arbuscular mycorrhizal fungal diversity and species dominance in a temperate soil with long-term conventional and low-input cropping systems

The aim of this work was to study the effect of long-term contrasting cropping systems on the indigenous arbuscular mycorrhizal fungal (AMF) spore populations in the soil of a field experiment located in western Finland. Conventional and low-input cropping systems were compared, each with two nutrient management regimes. The conventional cropping system with a non-leguminous 6-year crop rotation (barley–barley–rye–oat–potato–oat) was fertilized at either full (rotation A) or half (rotation B) the recommended rate. In the low-input cropping system, plant residues were returned to the plots either as such (rotation C) or composted (rotation D). In the rotation of this system, 1 year with barley was replaced by clover, and oat was cultivated mixed with pea. Thus, the 6-year rotation was barley–red clover–rye–oat + pea–potato–oat + pea. Each rotation was replicated three times, starting the 6-year rotation in three different years, these being designated point 1, point 2, and point 3, respectively. In the low-input system, biotite and rock phosphate were used to compensate for K and P in the harvested yield, while animal manure was applied at the start only. After 13 years, rotation points 1 and 3 were studied. Barley was the standing crop in all plots of rotation point 1, while oat and oat + pea were grown in rotations C and D, respectively. AMF spores were studied by direct extraction and by trapping, sampled on 15 June and 15 August. In addition, a special assay was designed for isolation of fast colonizing, dominating AMF. The cropping system did not significantly affect AMF spore densities, although the low-input cropping system with composted plant residues had the highest density with 44 spores on average and the conventional system with full fertilization 24 spores per 100 cm³ soil in the autumn samples. Species richness was low in the experimental area. Five Glomus spp., one Acaulospora, and one Scutellospora were identified at the species level. In addition to these, three unidentified Glomus spp. were found. Species richness was not affected by cropping system, rotation point, or their interactions. The Shannon–Wiener index of AMF spore distributions was significantly higher in the fully fertilized than in the half-fertilized conventional plots. Glomus claroideum was the most commonly identified single species in the experimental area. It occurred in all the cropping systems and their various rotation points, representing about 30% of the total number of identified spores. In August, G. claroideum accounted for as much as 45–55% of the total numbers of spores identified in the conventional system with halved fertilization. In contrast, Glomus mosseae occurred more commonly in June (26%) than in August (9%). A bioassay using roots as inoculum for isolation and culture of dominating AMF was successfully developed and yielded only G. claroideum. This indicates a high probability of being able to more generally identify, isolate, and culture fast colonizing generalist AMF for use as inoculants in agriculture and horticulture.     

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.     

Observations on chytridiaceous parasites of Phanerogams

Summary A reexamination of Physoderma maculare Wallroth the type species of the genus, on Alisma, confirms Clinton's (1902) account of the production of an epibiotic stage from resting spore zoospores. The latter on mature host tissue may also give rise to the endobiotic stage which bears resting spores. On seedlings, however, resting spore zoospores produce only epibiotic sporangia. The fungus could not successfully infect Sagittaria, a closely related host.Contribution No. 1152     

Viability of phytoplankton resting stages in the sediments of a coastal Swedish fjord

Viable diatom and dinoflagellate resting stages were recovered from sediments in Koljö Fjord on the west coast of Sweden. To determine the maximum survival time of buried resting stages, samples from sediment depths down to 50?cm were incubated at temperatures of 3, 10 and 18?°C. Sediment cores were dated by ²¹⁰Pb and the age of samples containing viable resting stages was determined using the constant rate of supply model. Dilution cultures of surface sediments allowed semiquantitative estimates of the potential seed bank. Dinoflagellate cysts from species such as Diplopsalis sp., Gymnodinium nolleri, Oblea rotunda and Protoceratium reticulatum were viable down to 15?cm depth, or 37 years old. Spores and resting cells of the diatoms Chaetoceros spp., Detonula confervacea and Skeletonema costatum were viable to over 40?cm depth, and may have been buried for many decades. The seed bank of living resting stages in surficial sediments was found to be rich (c. 57000 diatom resting stages g^?1 wet weight and c. 200 dinoflagellate cysts g^?1 wet weight), and the percentage of viable resting stages was higher for spore- and cyst-forming species. The oxygen-deficient sediments in Koljö Fjord appear to be a natural conservator of cell viability, a condition not easily simulated in laboratory studies. These results are ecologically important since spores and cysts are a repository of genetic material able to repopulate waters if resuspended and exposed to suitable light, temperature and nutrients.     

Post-glacial seasonal diatom record of the Mertz Glacier Polynya,East Antarctica

An ultra-high-resolution post-glacial laminated sediment record from Mertz Ninnis Trough, East Antarctic Margin (EAM), has been analysed using SEM backscattered electron imagery, secondary electron imagery and quantitative diatom abundance. Laminations are classified using visually dominant diatom species and terrigenous content. Four biogenic diatom ooze laminae types, one diatom-bearing terrigenous lamina type and one diatom-bearing terrigenous sub-lamina type have been identified. Diatom ooze lamina types comprise near-monogeneric Hyalochaete Chaetoceros spp. resting spore laminae, laminae characterised by Corethron pennatum, laminae characterised by Rhizosolenia spp. and mixed diatom assemblage laminae. Diatom-bearing terrigenous lamina and sub-lamina types comprise mixed diatom assemblage terrigenous laminae and sub-laminae characterised by Porosira glacialis resting spores. Formation of each of these lamina types is controlled by seasonal changes in nutrients, oceanographic regimes and the Mertz Glacier Polynya dynamics.