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.     

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.     

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 .     

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.     

Monterey Bay phytoplankton I. Seasonal cycles of phytoplankton assemblages

Netplankton collections from shallow-water stations in MontereyBay, California were examined for 1976 and 1977. The predominatingphytoplankton forms were neritic diatom species. Recurrent speciesgroups were identified using Fager's regroup analysis and theirdistributions related to hydrographic seasons. Neritic groupswere relatively more abundant during the Davidson Current andupweiling periods; however, many of the species were presentthroughout much of the year. The appearance of oceanic groupsand species was seasonal and associated with advective events. One neritic species group was of overwhelming numerical importancebut within the group conspicuous changes in relative abundanceof phytoplankton populations occurred seasonally among speciesfrom this group. This group appears to be composed of residentspecies and may be locally retained as vegetative cells or benthicresting stages. Species cycles in Monterey Bay may be regardedas largely the result of suc-cessional changes or cycles ofautochthonous species, while species introduced by adjectiveprocesses appear to be of minor importance. The study has suggested that the presence of species in a ‘seedpopulation’, either as vegetative cells or benthic restingstages, may be important as the growth rate of the species inunderstanding and predicting composition and relative abundancein coastal phytoplankton assemblages.     

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     

Increase in Si:N drawdown ratio due to resting spore formation by spring bloom-forming diatoms under Fe- and N-limited conditions in the Oyashio region

Resting spore formation and Si:N drawdown ratios were investigated under iron (Fe)- and nitrogen (N)-limited conditions using a unialgal culture of Thalassiosira nordenskioeldii and natural phytoplankton assemblages during the spring bloom in the Oyashio region. In the unialgal culture of T. nordenskioeldii, 20% and 100% of the cells formed resting spores under Fe- and N-limited conditions, respectively. The Si:N drawdown ratios were 2- and 14-fold higher in Fe- and N-limited conditions, respectively, compared to Fe- and N-sufficient conditions. At the start of the natural phytoplankton incubation, 18 among 47 identified diatom species were known resting spore-forming species. Approximately 15 common diatom species formed resting spores under Fe- and N-limited conditions. During the natural phytoplankton incubation, the percentage of the resting spores increased with time under both Fe- and N-limited conditions, reaching 25% and 40% of total diatom abundance, respectively. The Si:N drawdown ratios significantly increased with an increase in the contribution of resting spores in both the unialgal culture and natural phytoplankton incubations. These results suggest that if the bloom dominated by neritic, resting spore-forming diatom species decline by either Fe- or N-depletion, Si may be utilized preferentially to N in the upper mixed layer due to the formation of heavily silicified resting spores.     

In vitro formation of resting spores by the insect pathogenic fungus Entomophaga maimaiga

Field-collected resting spores (azygospores) of the fungal pathogen of Lymantria dispar (gypsy moth), Entomophaga maimaiga, have been used to release this biological control agent in areas where this pathogen is not established. We have found that E. maimaiga can produce resting spores in vitro using Grace's insect tissue culture medium (95%) plus fetal bovine serum (5%). The majority of spores become mature between 7 and 21 days after cultures are initiated. Spore production varies by fungal isolate; of 38 isolates tested, 10 produced no resting spores while 7 produced >1000 resting spores/ml. Resting spore production was not affected when isolates were mixed. Glycerol (used for fungal storage), trehalose, and selected amino acids each inhibited resting spore formation. Fetal bovine serum was required for spore production but the presence of >5% yielded lower resting spore densities. A large surface area:volume ratio (12.5 cm(2):ml versus 4.2 cm(2):ml) was required for abundant formation of resting spores. At present, resting spores have only been produced in small volumes with a maximum of 3 x 10(4) resting spores/ml.     

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。     

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。     

Entomophthora muscae resting spore formation in vivo in the host Delia radicum

The formation in vivo of Entomophthora muscae resting spores was investigated in the host, Delia radicum (cabbage root fly), by analysis of field data on the seasonal occurrence of E. muscae resting spores over 4 years. E. muscae resting spores in D. radicum were spherical with an average diameter of 39.4 microm, and the average numbers produced were estimated at 5.7 x 10(4) resting spores/female cadaver. Resting spores were found only after midsummer in D. radicum and almost exclusively in females. The proportion of infected females with resting spores was negatively correlated with average weekly day length after midsummer. We did not detect any significant year effect; thus, the results support the hypothesis that the photoperiod is the most important abiotic factor controlling E. muscae resting spore formation in D. radicum.     

Storage of Resting Spores of the Gypsy Moth Fungal Pathogen, Entomophaga maimaiga

The fungal pathogen, Entomophaga maimaiga causes epizootics in populations of the important North American forest defoliator gypsy moth ( Lymantria dispar ). Increasing use of this fungus for biological control is dependent on our ability to produce and manipulate the long-lived overwintering resting spores (azygospores). E. maimaiga resting spores undergo obligate dormancy before germination so we investigated conditions required for survival during dormancy as well as the dynamics of subsequent germination. After formation in the field during summer, resting spores were stored under various moisture levels, temperatures, and with and without soil in the laboratory and field. The following spring, for samples maintained in the field, germination was greatest among resting spores stored in plastic bags containing either moistened paper towels or sterile soil. Resting spores did not require light during storage to subsequently germinate. In the laboratory, only resting spores maintained with either sterile or unsterilized soil at 4°C (but not at 20 or -20°C) germinated the following spring, but at a much lower percentage than most field treatments. To further investigate the effects of relative humidity (RH) during storage, field-collected resting spores were placed at a range of humidities at 4°C. After 9.5 months, resting spore germination was highest at 58% RH and no resting spores stored at 88 or 100% RH germinated. To evaluate the dynamics of infections initiated by resting spores after storage, gypsy moth larvae were exposed to soil containing resting spores that had been collected in the field and stored at 4°C for varying lengths of time. No differences in infection occurred among larvae exposed to fall-collected soil samples stored at 4oC over the winter, versus soil samples collected from the same location the following spring. Springcollected resting spores stored at 4°C did not go into secondary dormancy. At the time that cold storage of soil containing resting spores began in spring, infection among exposed larvae was initiated within a few days after bringing the soil to 15°C. This same pattern was also found for spring-collected resting spore-bearing soil that was assayed after cold storage for 2-7 months. However, after 31-32 months in cold storage, infections started 14-18 days after soil was brought to 15°C, indicating a delay in resting spore activity after prolonged cold storage.     

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.     

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.     

GENETIC VARIABILITY WITHIN A POPULATION AND BETWEEN DIPLOID/HAPLOID TISSUE OF MACROCYSTIS PYRIFERA (PHAEOPHYCEAE)1

Multi-locus DNA fingerprints using an M13 probe were obtained for eight individuals of giant kelp Macrocystis pyrifera (L.) C. Ag. collected from Monterey Bay, California. For each individual, DNA was extracted from a diploid blade and from ca. 10⁹ haploid spores that were released from four to Jive sporophylls. Viable or swimming spores from one individual were pooled and referred to as a spore group. A total of 34 bands (4–19 kb) was detected in DNA fingerprints from the eight blades and eight spore groups, with individual blade or spore groups exhibiting 7–18 bands (mean = 12.6). One band (4.5 kb) was present in all 16 samples. Eight bands were detected in 11–14 of the 16 samples. Similarity indices were calculated for all pairwise comparisons of fingerprint bands among all possible combinations of blades and spore groups. Mean similarity indices for the eight blades (0.51, SE = 0.032) and spore groups (0.56, SE = 0.031) were significantly lower than for the eight comparisons of the blade and spore groups from a single individual (0.86, SE = 0.052). The data indicate that DNA fingerprints can be used to measure genetic variation within populations of M. pyrifera because variation of DNA fingerprints associated with meiotic products (spores) of a given individual is small relative to variation observed among individuals within the population. Additionally, fingerprint variation between diploid vegetative tissue and haploid meiotic products may be a measure of genetic change due to recombination or DNA turnover mechanisms.     

Diversity,abundance and expression of nitrite reductase (nirK)-like genes in marine thaumarchaea

Ammonia-oxidizing archaea (AOA) are widespread and abundant in aquatic and terrestrial habitats and appear to have a significant impact on the global nitrogen cycle. Like the ammonia-oxidizing bacteria, AOA encode a gene homologous to copper-containing nitrite reductases (nirK), which has been studied very little to date. In this study, the diversity, abundance and expression of thaumarchaeal nirK genes from coastal and marine environments were investigated using two mutually excluding primer pairs, which amplify the nirK variants designated as AnirKa and AnirKb. Only the AnirKa variant could be detected in sediment samples from San Francisco Bay and these sequences grouped with the nirK from Candidatus Nitrosopumilus maritimus and Candidatus Nitrosoarchaeum limnia. The two nirK variants had contrasting distributions in the water column in Monterey Bay and the California Current. AnirKa was more abundant in the epi- to mesopelagic Monterey Bay water column, whereas AnirKb was more abundant in the meso- to bathypelagic California Current water. The abundance and community composition of AnirKb, but not AnirKa, followed that of thaumarchaeal amoA, suggesting that either AnirKa is not exclusively associated with AOA or that commonly used amoA primers may be missing a significant fraction of AOA diversity in the epipelagic. Interestingly, thaumarchaeal nirK was expressed 10–100-fold more than amoA in Monterey Bay. Overall, this study provides valuable new insights into the distribution, diversity, abundance and expression of this alternative molecular marker for AOA in the ocean.     

The effect of eight fungicides on germination of Conidiobolus obscurus resting spores

Conidiobolus obscurus resting spore germination was greater than 60% after 21 days incubation in 700 ppm concentrations of Rhodiasoufre, Pelt 44, Dithane M-45, Plantvax, and Saprol. No germ tubes were produced under the same treatment with Bavistine, Quin 20, or Benlate at the same dosage. Germination in distilled water was 85%. After only 24 hr contact with Quin 20 (700 ppm) followed by a distilled water wash, germination was not reduced. A higher ability to penetrate the spore wall may account for greater fungicide activity. Resting spores were found to be more resistant to certain fungicides than conidia; however, no general rule could be applied.     

Detection and quantification of airborne conidia of Fusarium circinatum, the causal agent of pine pitch canker, from two California sites by using a real-time PCR approach combined with a simple spore trapping method

Pinus radiata (Monterey pine), a tree native to coastal California and Mexico, is widely planted worldwide for timber production. A major threat to Monterey pine plantations is the fungal disease pine pitch canker, caused by Fusarium circinatum (Hypocreales). We present a novel trapping approach using filter paper in combination with a rapid molecular method to detect the presence of inoculum in the air. The assay is also useful for diagnosing the presence of the pathogen on plants. The test is based on the F. circinatum specific primer pair CIRC1A-CIRC4A, which amplifies a 360-bp DNA fragment in the intergenic spacer region of the nuclear ribosomal operon. Real-time PCR was used to calculate the number of fungal spores present in each reaction mixture by comparing the threshold cycle (Ct) of unknown spore samples to the Ct values of standards with known amounts of F. circinatum spores. The filter paper method allows prolonged and more sensitive spore sampling in the field compared to traditional traps using petri dishes filled with selective medium. A field test at two sites in coastal California infested with pine pitch canker was carried out during the summer and fall of 2002. Spore counts were in the range of ca. 1 x 10(3) to ca. 7 x 10(5)/m(2), with the highest spore counts in the fall, suggesting a seasonal fluctuation.     

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.     

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.