130 "Plantae" Emended Based on the Common History of the Primary Plastid Endosymbiosis

Travertine terraces have been deposited by calcareous hot springs in Yellowstone from as early as 365,000 years to the present. Most of these porous and non-porous CaCO3 rocks (old or new) contain a 1–2 mm thick greenish band about 1–3 mm below the upper surface. These bands are composed of cyanobacteria and, sometimes, unicellular green algae. Although some moisture may be retained for much of the year, all undergo freezing in winter and desiccation in summer. DGGE (denaturing gradient gel electrophoresis), with subsequent 16S rDNA sequence analyses of bands, has shown that relatively few phylotypes of cyanobacteria are present, but some occur in travertine of very different ages, indicating secondary establishment of the communities. Clonal cultures of predominant types have also been established and sequenced. All those tested are able to survive extreme desiccation. Preliminary sequence analyses of cultures show that some strains are nearly identical to known cyanobacterial strains while others show little similarity. One sequence is 100% identical to the cyanobacterium Cyanobium gracile. This cyanobacterium is known to be distributed worldwide in lakes and brackish seas, but not in a cryptoendolithic environment. Another sequence shows 99% identity to two cyanobacteria isolated from Antarctic freshwater ponds. Both Antarctic ponds and Yellowstone travertine are environments in which adaptations for desiccation and/or freezing tolerance could be crucial. The lack of ecological similarity among some of these strains indicates that genes other than 16S rDNA must be used for differentiation. These results will be discussed along with the ecology of travertine habitats.     

Characterisation of Antarctic cyanobacteria and comparison with New Zealand strains

Cyanobacterial mats are common in Antarctic lakes, ponds and on moist soils. The species comprising these mats have adapted to tolerate extreme conditions (e.g. high salinities and UV radiation, freezing and extended periods of darkness). In this study, cyanobacterial mats were collected from shallow melt-water ponds in Pyramid Trough in Southern Victoria Land, Antarctica. Eight strains were isolated and characterised by morphological and molecular (16S rRNA gene sequences) techniques and their fatty acid methyl ester (FAME) and lipid class profiles determined. These data were compared to parallel information obtained from cyanobacterial cultures isolated from New Zealand. In general, the morphological and molecular characterisation complemented each other, and the Antarctic strains identified belonged to the orders: Oscillatoriales (six), Nostocales (one) and Chroococcales (one). Two of the Antarctic strains (CYN67 and CYN68) showed low similarity (<96% 16S rRNA gene sequence) when compared to other cultured cyanobacteria. The fatty acid (FA) profiles from the Antarctic and New Zealand strains shared many similarities with palmitic (C16:0), stearic (C18:0) and oleic acid (C18:1n-9) most abundant. In contrast, the lipid class analysis differed among geographic locations with Antarctic strains containing higher amounts of hydrocarbons and eicosapentaenoic and hexadecatrienoic acids.     

Response of Cyanobacteria and Algae from Antarctic Wetland Habitats to Freezing and Desiccation Stress

Antarctic wetlands are characterized by the presence of liquid water during short austral summer. Filamentous cyanobacteria are often dominant there and are exposed to severe conditions, of which the changes in the desiccation–rehydration and freeze–thaw cycles are two of the most stressful. Vigor, after freezing and desiccation, was laboratory tested in cyanobacterial and algal strains from wetland habitats collected in maritime and continental Antarctica. Whereas minor sub-zero temperatures (−4°C), demonstrating summer diurnal freeze–thaws did not cause significant damage on either cyanobacteria or algae, low sub-zero temperatures (−40, −100, −196°C), demonstrating annual winter freeze, caused little harm to cyanobacteria, but was fatal for more than 50% of the population of algae. Freezing and desiccation tolerance of these strains was compared using multiregression methods: cyanobacteria from continental Antarctica were significantly more tolerant to low sub-zero temperatures than similar strains from maritime Antarctica (P = 0.026; F = 3.66); and cyanobacteria from seepages habitat were less tolerant to freezing and desiccation than cyanobacteria from other wetlands (P = 0.002; F = 5.69).     

Endolithic photosynthetic communities within ancient and recent travertine deposits in Yellowstone National Park

Molecular and culture based methods were used to survey endolithic, photosynthetic communities from hot spring-formed travertine rocks of various ages, ranging from<10 to greater than 300,000 years. Much of this travertine contained a 1-3-mm-thick greenish band composed mainly of cyanobacteria 1-5 mm below the rock surface. The travertine rocks experienced desiccation in summer and freezing in winter. A total of 83 environmental 16S rRNA gene sequences were obtained from clone libraries and denaturing gradient gel electrophoresis. Small subunit rRNA gene sequences and cell morphology were determined for 36 cyanobacterial culture isolates from these samples. Phylogenetic analysis showed that the 16S rRNA gene sequences fell into 15 distinct clusters, including several novel lineages of cyanobacteria.     

Formation of laminate travertines at bagno vignone,Italy

Travertine at Bagno Vignoni, Tuscany, is deposited from thermal CO₂‐rich water up‐welling from an artificial pool, Bagno Grande, which contains a thick benthic mat of cyanobacteria. The travertine mound, over 40 m in height, consists of calcite feather crystals and micrite. Over much of the mound, but particularly in the lower part, the travertine is finely banded, with pale bands of feather crystals alternating with dark bands containing micrite. Deposition rate measurements showed that the bands were formed daily, with the slightly narrower and darker bands formed at night. These nocturnal bands also contained wind‐borne dust. It is postulated that the dust caused heterogeneous nucleation of micrite and reduced the development of feather crystals. The deposition rate of travertine ranged from 127 to 348 μm/day and was highest at the top of the mound. Water analysis conducted over 27 h demonstrated a pronounced diurnal variation in the dissolved carbon dioxide of Bagno Grande, consistent with the photosynthetic activity of the benthic cyanobacteria. This variation could be traced to the base of the travertine mound but was not alone responsible for the higher deposition rate during the day. Water temperature on the mound also increased during the day and would have aided CO₂ evasion. Photosynthetic activity on the mound itself appeared to have far less influence on the carbon dioxide flux than Bagno Grande. The upper mound supported unlithified mats of the cyanobacteria Lyngbya (Phormidium) laminosum and Spirulina labyrinthiformis. These mats sometimes possessed fine travertine laminae resulting from cyanobacterium phototaxis and may have been responsible for some lamination in the upper mound. The CO₂ flux measurements indicated that overall, atmospheric evasion of carbon dioxide was more significant to travertine deposition than photosynthetic activity.     

EVOLUTIONARY RELATIONSHIPS OF CULTIVATED ANTARCTIC OSCILLATORIANS (CYANOBACTERIA)

The evolutionary relationships of cultivated psychrophilic and psychrotolerant polar oscillatorians were examined, based on small subunit rDNA sequences. Psychrophilic oscillatorians from the Antarctic were affiliated in one well-supported clade, which also includes two Arctic strains. Two of the Antarctic psychrophiles contain an 11-nucleotide insertion that is identical to, and previously only described in, an Arctic oscillatorian. The psychrotolerant phenotype, conversely, has arisen multiple times in the cyanobacterial lineage, and psychrotolerant strains are sometimes most closely related to organisms of temperate latitudes. These findings support the hypothesis that oscillatorians in both polar regions originated from more temperate species. Furthermore, morphological designations of these filamentous cyanobacteria often do not have phylogenetic significance.     

Molecular characterization of cyanobacterial diversity in a shallow eutrophic lake

We have studied the diversity of pelagic cyanobacteria in Lake Loosdrecht, The Netherlands, through recovery and analysis of small subunit ribosomal RNA gene sequences from lake samples and cyanobacterial isolates. We used an adapted protocol for specific amplification of cyanobacterial rDNA for denaturing gradient gel electrophoresis (DGGE) analysis. This protocol enabled direct comparison of cyanobacterial community profiles with overall bacterial profiles. The theoretical amplification specificity of the primers was supported by sequence analysis of DNA from excised DGGE bands. Sequences recovered from these bands, in addition to sequences obtained by polymerase chain reaction (PCR) and cloning from lake DNA as well as from cyanobacterial isolates from the lake, revealed a diverse consortium of cyanobacteria, among which are representatives of the genera Aphanizomenon, Planktothrix, Microcystis and Synechococcus. One numerically important and persistent cyanobacterium in the lake, Prochlorothrix hollandica, appeared to co-occur with an unknown but related species. However, the lake is dominated by filamentous species that originally have been termed 'Oscillatoria limnetica-like'. We show that this is a group of several related cyanobacteria, co-occurring in the lake, which belong to the Limnothrix/Pseudanabaena group. The available variation among the coexisting strains of this group can explain the persistent dominance of the group under severe viral pressure.     

Freezing and desiccation injury resistance in the filamentous green alga Klebsormidium from the Antarctic, Arctic and Slovakia

The freezing and desiccation tolerance of 12 Klebsormidium strains, isolated from various habitats (aeroterrestrial, terrestrial, and hydro-terrestrial) from distinct geographical regions (Antarctic — South Shetlands, King George Island, Arctic — Ellesmere Island, Svalbard, Central Europe — Slovakia) were studied. Each strain was exposed to several freezing (−4°C, −40°C, −196°C) and desiccation (+4°C and + 20°C) regimes, simulating both natural and semi-natural freeze-thaw and desiccation cycles. The level of resistance (or the survival capacity) was evaluated by chlorophyll a content, viability, and chlorophyll fluorescence evaluations. No statistical differences (Kruskal-Wallis tests) between strains originating from different regions were observed. All strains tested were highly resistant to both freezing and desiccation injuries. Freezing down to −196°C was the most harmful regime for all studied strains. Freezing at −4°C did not influence the survival of studied strains. Further, freezing down to −40°C (at a speed of 4°C/min) was not fatal for most of the strains. RDA analysis showed that certain Antarctic and Arctic strains did not survive desiccation at +4°C; however, freezing at −40°C, as well as desiccation at +20°C was not fatal to them. On the other hand, other strains from the Antarctic, the Arctic, and Central Europe (Slovakia) survived desiccation at temperatures of +4°C, and freezing down to −40°C. It appears that species of Klebsormidium which occupy an environment where both seasonal and diurnal variations of water availability prevail, are well adapted to freezing and desiccation injuries. Freezing and desiccation tolerance is not species-specific nor is the resilience only found in polar strains as it is also a feature of temperate strains. Presented at the International Symposium Biology and Taxonomy of Green Algae V, Smolenice, June 26–29, 2007, Slovakia. This paper is dedicated to the memory of the late Dr. Bohuslav Fott (1908–1976), Professor of Botany at the Charles University in Prague, to mark the centenary of his birth.     

Lithic cyanobacterial communities in the polyextreme Sahara Desert: implications for the search for the limits of life

The hyperarid Sahara Desert presents extreme and persistent dry conditions with a limited number of hours during which the moisture availability, temperature and light allow phototrophic growth. Some cyanobacteria can live in these hostile conditions by seeking refuge under (hypolithic) or inside (endolithic) rocks, by colonizing porous spaces (cryptoendoliths) or fissures in stones (chasmoendoliths). Chroococcidiopsis spp. have been reported as the dominant or even the only phototrophs in these hot desert lithic communities. However, the results of this study reveal the high diversity of and variability in cyanobacteria among the sampled habitats in the Sahara Desert. The chasmoendolithic samples presented high coccoid cyanobacteria abundances, although the dominant cyanobacteria were distinct among different locations. A high predominance of a newly described cyanobacterium, Pseudoacaryochloris sahariense, was found in hard, compact, and more opaque stones with cryptoendolithic colonization. On the other hand, the hypolithic samples were dominated by filamentous, non-heterocystous cyanobacteria. Thermophysiological bioassays confirmed desiccation and extreme temperature tolerance as drivers in the cyanobacterial community composition of these lithic niches. The results of the present study provide key factors for understanding life strategies under polyextreme environmental conditions. The isolated strains, especially the newly described cyanobacterium P. sahariense, might represent suitable microorganisms in astrobiology studies aimed at investigating the limits of life.     

Enrichment culture and microscopy conceal diverse thermophilic Synechococcus populations in a single hot spring microbial mat habitat.

Recent molecular studies have shown a great disparity between naturally occurring and cultivated microorganisms. We investigated the basis for disparity by studying thermophilic unicellular cyanobacteria whose morphologic simplicity suggested that a single cosmopolitan species exists in hot spring microbial mats worldwide. We found that partial 16S rRNA sequences for all thermophilic Synechococcus culture collection strains from diverse habitats are identical. Through oligonucleotide probe analysis and cultivation, we provide evidence that this species is strongly selected for in laboratory culture to the exclusion of many more-predominant cyanobacterial species coexisting in the Octopus Spring mat in Yellowstone National Park. The phylogenetic diversity among Octopus Spring cyanobacteria is of similar magnitude to that exhibited by all cyanobacteria so far investigated. We obtained axenic isolates of two predominant cyanobacterial species by diluting inocula prior to enrichment. One isolate has a 16S rRNA sequence we have not yet detected by cloning. The other has a 16S rRNA sequence identical to a new cloned sequence we report herein. This is the first cultivated species whose 16S rRNA sequence has been detected in this mat system by cloning. We infer that biodiversity within this community is linked to guild structure.     

Enrichment culture and microscopy conceal diverse thermophilic Synechococcus populations in a single hot spring microbial mat habitat

Recent molecular studies have shown a great disparity between naturally occurring and cultivated microorganisms. We investigated the basis for disparity by studying thermophilic unicellular cyanobacteria whose morphologic simplicity suggested that a single cosmopolitan species exists in hot spring microbial mats worldwide. We found that partial 16S rRNA sequences for all thermophilic Synechococcus culture collection strains from diverse habitats are identical. Through oligonucleotide probe analysis and cultivation, we provide evidence that this species is strongly selected for in laboratory culture to the exclusion of many more-predominant cyanobacterial species coexisting in the Octopus Spring mat in Yellowstone National Park. The phylogenetic diversity among Octopus Spring cyanobacteria is of similar magnitude to that exhibited by all cyanobacteria so far investigated. We obtained axenic isolates of two predominant cyanobacterial species by diluting inocula prior to enrichment. One isolate has a 16S rRNA sequence we have not yet detected by cloning. The other has a 16S rRNA sequence identical to a new cloned sequence we report herein. This is the first cultivated species whose 16S rRNA sequence has been detected in this mat system by cloning. We infer that biodiversity within this community is linked to guild structure.     

Phylogenetic relationships between geographically separate Phormidium cyanobacteria: is there a link between north and south polar regions?

We present a phytogeographical comparison between polar (Arctic and Antarctic) and non-polar strains of the cyanobacterial genus Phormidium, which plays a key role in Arctic and Antarctic ecosystems as primary producer. A total of 26 Phormidium strains were studied using a polyphasic approach, 18 from Arctic (Svalbard, Ellesmere Island and Scandinavian Arctic—Abisko) and Antarctic (Antarctic Peninsula—King George and James Ross Island) regions, and 8 from temperate sites (mostly situated in Central Europe). A phylogenetic tree was constructed and compared with similar 16S rRNA sequences retrieved from Genbank. Within the Phormidium autumnale cluster, genetic similarity of 16S rDNA was more related to geographical proximity of strain origin than to morphological similarity. No genetic identity of Phormidium strains from north and south polar regions was found. The cluster Phormidium autumnale apparently belongs to generic entities in which geographical limitation plays a prominent role. However, the cyanobacterial strains found in Europe suggest that the distribution areas of some Phormidium cyanobacteria overlap. The Phormidium autumnale cluster is evidently a very characteristic type and represents an isolated clade within the traditional genus Phormidium. According to morphological features and the structure of trichomes, it is most similar and thus probably belongs to the genus Microcoleus.     

An assessment of biopotential of three cyanobacterial isolates from Antarctic for carotenoid production

Specific growth rates and carotenoid contents of three Antarctic and tropical strains of cyanobacteria viz. Anabaena sp., Phormidium sp. and Nostoc sp. were compared in batch and mass cultures to assess bio-potential of Antarctic strains for cost-effective carotenoid production. Antarctic strains though exhibited slightly lower specific growth rates, but contained higher carotenoid contents (per unit dry wt.), than tropical strains. Modification of normal composition of BG-11 culture medium, by altering nitrogen and carbon sources resulted in 25-38% increase in carotenoid content in both types of strains. Mass-culture in indoor and semi-outdoor bio-reactors resulted in 39-113% higher carotenoid content in Antarctic strains, compared to their respective tropical strains. The observations suggest that Antarctic cyanobacteria may have potential as superior strains for maximizing the yield of carotenoids.     

The effects of freezing and desiccation on photosynthesis and survival of terrestrial Antarctic algae and cyanobacteria

Summary The effects of low temperatures, freezing and desiccation on a cyanobacterium (Phormidium) and an alga (Prasiola) from terrestrial Antarctic habitats were investigated. Net photosynthesis per unit dry weight, measured by gas exchange, and the vital stain Auramine O were used to monitor recovery from stress. Photosynthetic rates by Prasiola were an order of magnitude higher than those by Phormidium, although both continued photosynthesis at sub-zero temperatures. Prasiola survived freezing more readily, but in both cases survival was dependant upon the prevailing light conditions and the presence/absence of free water. Phormidium readily survived desiccation, whereas high mortality occurred in Prasiola, particularly at high light intensities. The results obtained are discussed in relation to the habitat and ecology of the organisms.     

The environmental physiology of Antarctic terrestrial nematodes: a review

The environmental physiology of terrestrial Antarctic nematodes is reviewed with an emphasis on their cold-tolerance strategies. These nematodes are living in one of the most extreme environments on Earth and face a variety of stresses, including low temperatures and desiccation. Their diversity is low and declines with latitude. They show resistance adaptation, surviving freezing and desiccation in a dormant state but reproducing when conditions are favourable. At high freezing rates in the surrounding medium the Antarctic nematode Panagrolaimus davidi freezes by inoculative freezing but can survive intracellular freezing. At slow freezing rates this nematode does not freeze but undergoes cryoprotective dehydration. Cold tolerance may be aided by rapid freezing, the production of trehalose and by an ice-active protein that inhibits recrystallisation. P. davidi relies on slow rates of water loss from its habitat, and can survive in a state of anhydrobiosis, perhaps aided by the ability to synthesise trehalose. Teratocephalus tilbrooki and Ditylenchus parcevivens are fast-dehydration strategists. Little is known of the osmoregulatory mechanisms of Antarctic nematodes. Freezing rates are likely to vary with water content in Antarctic soils. Saturated soils may produce slow freezing rates and favour cryoprotective dehydration. As the soil dries freezing rates may become faster, favouring freezing tolerance. When the soil dries completely the nematodes survive anhydrobiotically. Terrestrial Antarctic nematodes thus have a variety of strategies that ensure their survival in a harsh and variable environment. We need to more fully understand the conditions to which they are exposed in Antarctic soils and to apply more natural rates of freezing and desiccation to our studies.Communicated by: I.D. Hume     

中国淡水共球藻纲新记录属种——土佐牧野藻(Makinoella tosaensis Okada)

报道分别从湖北省武汉市内和云南省西双版纳小水池中分离培养的两株绿藻,对其进行了形态和18S r DNA基因序列分析,编号分别为FACHB-1783和FACHB-1784。这两株绿藻具独特的四边形群体形态,通常为4或16个细胞,细胞为宽椭圆形至不规则卵圆形、细胞壁两端无增厚,叶绿体多数、片状,具蛋白核。结合形态和分子系统发育分析,确定这两株绿藻为我国1种淡水共球藻纲新记录属种——土佐牧野藻(Makinoella tosaensis Okada)。基于18S r DNA基因的系统发育研究表明这两株绿藻与分离自韩国的土佐牧野藻基因序列相似度可达99.6%~99.9%,并且以较高的支持值与土佐牧野藻聚在一起。     

Effect of exogenous extracellular polysaccharides on the desiccation and freezing tolerance of rock-inhabiting phototrophic microorganisms

Two major stresses that threaten rock-inhabiting microbial communities are desiccation and freezing; both result in a loss of liquid water in the cells. The mechanisms necessary to tolerate these extremes may be similar, but are not well understood. In both cases extracellular polysaccharides (EPS) seem to play an important role. This study examines whether the EPS released by a rock-inhabiting phototroph can have a protective effect on other members of similar and neighboring microbial communities. This interaction was modeled by adding EPS isolated from the cryptoendolithic cyanobacterium Nostoc sp. to cells of the cryptoendolithic green alga Chlorella sp. and to cells of the epilithic cyanobacterium Chroococcidiopsis sp. The cells were then subjected to desiccation and freezing and the survival rates were determined by vital staining, using membrane integrity as a measure of viability. The results clearly demonstrate the importance of exogenous EPS in the desiccation tolerance of both species, while mixed results were found for the freezing trials.     

Cyanobacterial construction of hot spring siliceous stromatolites in Yellowstone National Park

Living stromatolites growing in a hot spring in Yellowstone National Park are composed of silica-encrusted cyanobacterial mats. Two cyanobacterial mat types grow on the stromatolite surfaces and are preserved as two distinct lithofacies. One mat is present when the stromatolites are submerged or at the water-atmosphere interface and the other when stromatolites protrude from the hot spring. The lithofacies created by the encrustation of submerged mats constitutes the bulk of the stromatolites, is comprised of silica-encrusted filaments, and is distinctly laminated. To better understand the cyanobacterial membership and community structure differences between the mats, we collected mat samples from each type. Molecular methods revealed that submerged mat cyanobacteria were predominantly one novel phylotype while the exposed mats were predominantly heterocystous phylotypes (Chlorogloeopsis HTF and Fischerella). The cyanobacterium dominating the submerged mat type does not belong in any of the subphylum groups of cyanobacteria recognized by the Ribosomal Database Project and has also been found in association with travertine stromatolites in a Southwest Japan hot spring. Cyanobacterial membership profiles indicate that the heterocystous phylotypes are 'rare biosphere' members of the submerged mats. The heterocystous phylotypes likely emerge when the water level of the hot spring drops. Environmental pressures tied to water level such as sulfide exposure and possibly oxygen tension may inhibit the heterocystous types in submerged mats. These living stromatolites are finely laminated and therefore, in texture, may better represent similarly laminated ancient forms compared with more coarsely laminated living marine examples.     

Biliproteins and phycobilisomes from cyanobacteria and red algae at the extremes of habitat

This review considers the properties of biliproteins from cyanobacteria and red algae that grow in extreme habitats. Three situations are presented: cyanobacteria that grow at high temperatures; a red alga that grows in acidic conditions at high temperature; and an Antarctic red alga that grows in the cold in dim light conditions. In particular, the properties of their biliproteins are compared to those from organisms from more usual environments. C-phycocyanins from two cyanobacteria able to grow at high temperatures are found to differ in their stabilities when compared to C-phycocyanin from mesophilic algae. They differ in opposite ways, however. One is more stable to dissociation than the mesophilic protein, and the other is more easily dissociated at low temperatures. The thermophilic proteins resist thermal denaturation much better than the mesophilic proteins. The most thermophilic cyanobacterium has a C-phycocyanin with a unique blue-shifted absorption maximum which does not appear to be part of the adaptation of the cyanobacterium to high temperature. The C-phycocyanin from the high-temperature red alga is able to resist dissociation better than mesophilic C-phycocyanins. Electron micrographs show the phycobilisomes of these algae. The Antarctic alga grows under ice at some distance down the water column. Its R-phycoerythrin has a novel absorption spectrum that gives the alga an improved ability to harvest blue light. This may enhance its survival in its light-deprived habitat.     

Phenotypic and genotypic characterization of multiple strains of the diazotrophic cyanobacterium, Crocosphaera watsonii, isolated from the open ocean

Diazotrophic cyanobacteria have long been recognized as important sources of reduced nitrogen (N) and therefore are important ecosystem components. Until recently, species of the filamentous cyanobacterium Trichodesmium were thought to be the primary sources of fixed N to the open ocean euphotic zone. It is now recognized that unicellular cyanobacteria are also important contributors, with members of the oligotrophic genus Crocosphaera being the only cultured examples. Herein we genetically and phenotypically characterize 10 strains isolated from the tropical Atlantic and North Pacific Oceans, and show that although all of the strains are highly similar at the genetic level, with the internal transcribed sequence (ITS) region sequence varying by ∼2 bp on average, there are many unexpected phenotypic differences between the isolates (e.g. cell size, temperature optima and range, extracellular material excretion and variability in rates of nitrogen fixation). However based on the observed sequence similarity, we propose that all of these isolates are members of the genus Crocosphaera (type strain Crocosphaera watsonii WH8501), and that the phenotypic diversity we see may reflect ecologically important variation relevant for modelling N₂ fixation in the oligotrophic ocean.