Phylogeography of the genus <Emphasis Type="Italic">Ulva</Emphasis> (Ulvophyceae,Chlorophyta), with special reference to the Japanese freshwater and brackish taxa

The nuclear-encoded ITS and associated 5.8S rDNA regions were sequenced for 72 specimens of Ulva collected from 44 rivers across Japan, including U. prolifera Müller from the Shimanto River, Kochi Prefecture, as well as 26 samples originally identified as U. linza L. from 20 coastal marine areas. Sequence data revealed that the samples fall into six distinct clades: the U. flexuosa Wulfen clade (2 samples), the Ulva linza-procera-prolifera (LPP) complex clade (75 samples), Ulva sp. 1 clade (3 samples), Ulva sp. 2 clade (7 samples), Ulva sp. 3 clade (4 samples) and Ulva sp. 4 clade (7 samples). The LPP complex contained a mixture of 26 samples collected from seashores and 49 samples obtained from rivers, including U. prolifera from the Shimanto River, and GenBank data for U. linza and U. procera Ahlner. The samples of the LPP complex differed by only 0–7 substitutions (0–1.149%). Subsequent phylogeographic analyses of the LPP complex based on the 5S rDNA spacer region revealed the presence of two further groupings: a group including 22 strictly marine littoral U. linza samples and a U. prolifera group composed of a mixture of 4 marine samples and all 49 river samples. The monophyly of all river samples indicates that adaptation to low salinity might have occurred only once in the evolutionary history of the LPP complex.     

Morphological and molecular characterization of free-floating and attached green macroalgae Ulva spp. in the Yellow Sea of China

During the summer of 2008 and 2009, massive algal blooms repeatedly broke out in the Yellow Sea of China. These were undoubtedly caused by the accumulations of one or more species in the macroalgal genus Ulva. In previous reports, morphological observation indicated that the species involved in this phenomenon is Ulva prolifera but molecular analyses indicated that the species belongs to an Ulva linza–procera–prolifera (LPP) clade. Correct identification of the bloom species is required to understand and manage the blooms, but the taxonomic status of the bloom species remains unclear. In the current study, the taxonomic status of 22 selected specimens from the Yellow Sea was assessed by using both morphological and molecular (ITS and rbcL sequences) data. In addition, 5S rDNA analyses were performed for those samples clustering in the LPP clade, and phylogenetic tree and ribotype analyses were constructed for determining the possible origin of the bloom. Three free-floating and two attached Ulva species were distinguished and described: Ulva compressa Linnaeus and Ulva pertusa Kjellman were found in free-floating samples; U. linza Linnaeus was found on rocks; and U. prolifera O.F. Müller was found in both habitats. Diversity in free-floating Ulva of the Yellow Sea appears to be greater than previously thought. The dominant free-floating Ulva species, U. prolifera, was not closely related to local populations attached to rocks but was closely related to populations from Japan.     

Seasonal variation of dominant free-floating and attached Ulva species in Rudong coastal area,China

In this paper, species compositions and seasonal variations of attached Ulva species on Porphyra aquaculture rafts and free floating Ulva species at Rudong coastal area, Jiangsu Province of China were investigated during 2010–2011. Based on the sequences analysis of nuclear-encoded ITS (including 5.8S rDNA regions) and 5S rDNA spacer regions, dominant species of both attached and free-floating Ulva samples were identified as Ulva compressa, Ulva linza, Ulva prolifera and Ulva flexuosa. Phylogenetic tree based on sequences of ITS and 5S rDNA spacer regions for attached and free-floating Ulva species was constructed, respectively. Species compositions of the Ulva population attached on aquaculture rafts varied with seasons, and U. prolifera was only found on aquaculture rafts in March 2011 during the 2010–2011 Porphyra yezoensis cultivation season, which had the same sequences of ITS and 5S rDNA spacer regions as that of the dominant species bloomed in the Yellow Sea of China in 2008. Dominant species of the free-floating Ulva population at the early stage of the green tide were U. compressa, U. flexuosa, and U. linza. Free-floating U. prolifera appeared in the middle of May, 2011. ITS sequence similarity rates of U. compressa and U. flexuosa between the attached and free-floating species were 100%. And ITS and 5S rDNA spacer sequences of the attached and the free-floating U. prolifera population also showed no differences. Further study showed that there were two types of free-floating U. prolifera population (Type 5S-A and Type 5S-B) based on 5S rDNA spacer sequences. The present study would provide some useful information for clarifying the outbreak mechanism of green tides occurred in the Yellow Sea, China.     

INVESTIGATIONS INTO SOUTHERN AUSTRALIAN ULVA (ULVOPHYCEAE,CHLOROPHYTA) TAXONOMY AND MOLECULAR PHYLOGENY INDICATE BOTH COSMOPOLITANISM AND ENDEMIC CRYPTIC SPECIES1

Appreciation of the true species diversity of the genus Ulva in Australian waters has been blinkered by the unproved assumption that its representatives there are largely cosmopolitan. As species of Ulva are some of the longest‐standing and most widely reported taxa of macroalgae, the presumption that they are worldwide in distribution has led to most Australian members being equated with species originally described from extra‐Australian type localities. Ulva species can be notoriously difficult to identify due to the few and often variable characters on which classical taxonomic studies focus so that names of specimens in hand, as well as names appearing in historical distribution records, are frequently difficult or impossible to verify. The combination of morphological and molecular analyses, the latter involving both nuclear (internal transcribed spacer [ITS]) and plastid (rbcL) markers, is critically important in taxonomic studies of the genus and has here been applied to selected Ulva populations from mostly cool‐temperate southern Australian localities. It has been determined that habit‐ and anatomy‐based keys of standard taxonomic literature are largely adequate for assigning species names based on classical concepts, but they often obscure a number of cryptic and pseudocryptic species that do not conform to extra‐Australian populations of the same designation, as indicated by the corresponding molecular data. Here, we present six species (Ulva australis Aresch., U. compressa Forssk., U. fasciata Delile, U. intestinalis L., U. laetevirens Aresch., U. tanneri H. S. Hayden et J. R. Waaland) for which anatomical and molecular data were congruent with both classical concepts and GenBank accession data and confirm these as cosmopolitan taxa in Australia. We also present six putative species designations based on anatomy [U. clathrata (Roth) C. Agardh, U. flexuosa Wulfen, U. linza L., U. prolifera O. F. Müll., U. stenophylla Setch. et N. L. Gardner, U. brisbanensis sp. nov.] that are inconsistent with molecular data, suggesting novel or cryptic taxa not represented in GenBank.     

Ulva diversity in the Yellow Sea during the large‐scale green algal blooms in 2008–2009

In the Yellow Sea of China, large‐scale green tides have broken out for three consecutive years from 2007 to 2009. As part of the efforts to localize the algal source, two cruises were conducted in the early stage and the outbreak stage of the bloom in 2009. We analyzed the morphological and genetic diversity of drifting Ulva specimens and culture‐derived isolates from seawater sampled in different localities. For phylogenetic analyses, the nuclear encoded ribosomal DNA internal transcribed spacer region (ITS nrDNA) and the plastid encoded large subunit of ribulose‐1, 5‐bisphosphate carboxylase/oxgenase gene (rbcL) were used. Our molecular and morphological data indicate that the dominant free‐floating Ulva species in 2008 and 2009 possibly belonged to a single strain of the U. linza‐procera‐prolifera (LPP) clade. The ITS sequences from bloom‐forming algal samples with dense branches were identical to those from U. linza‐like specimens without branches derived from the Yellow Sea. Microscopic individuals of the dominant Ulva strain were detected in eight stations, revealing that spore dispersal in the water helped to enlarge biomass in the water during the outbreak stage of green tide in the Yellow Sea.     

Genetic and ecophysiological diversity of Cladophora (Cladophorales,Ulvophyceae) in various salinity regimes

Although Cladophora species frequently appear in brackish environments, their genetic diversity, phenological patterns and physiological properties have not been well investigated in these environments. Cladophora is distributed throughout Mikata‐goko, an area consisting of five coastal lakes that are directly or indirectly connected to the sea, resulting in a salinity gradient ranging from fully marine to freshwater. To elucidate genetic and ecological variation in Cladophora, we monthly compared ribosomal internal transcribed spacer sequences of Cladophora specimens across six study sites characterized by different salinity regimes for a year. A total of 12 ribotypes were detected and assigned to six species, four of which were restricted to the marine habitat. Ribotype I of C. vagabunda (L.) Hoek was distributed in high‐salinity brackish waters (mean salinity ≤13 psu; maximum salinity ≤31 psu), whereas C. vagabunda ribotype II and C. glomerata were abundant in low‐salinity brackish waters (mean salinity ≤5 psu; maximum salinity ≤10 psu). Although Cladophora albida (Nees) Kütz. and C. glomerata (L.) Kütz. were collected during all four seasons, C. laetevirens (Dillwyn) Kütz., C. oligocladoidea Hoek and Chihara, C. opaca Sakai and C. vagabunda displayed marked seasonal variation. Culture experiments revealed that optimal salinity conditions for vegetative growth vary among ribotypes collected from different salinity regimes, suggesting that Cladophora distributions are controlled by ribotype‐specific ecophysiological adaptations. In contrast, temperature optima and tolerance were similar among ribotypes showing different seasonalities, and thus their phenologies may be controlled by other environmental factors or biotic conditions, such as reproductive maturity and spore germination.     

Discrimination of the common macroalgae (Ulva and Blidingia) in coastal waters of Yellow Sea,northern China,based on restriction fragment-length polymorphism (RFLP) analysis

Since 2007, reoccurring large-scale green algae blooms have caused deleterious effects to the estuarine ecosystem of Yellow Sea, northern China and subsequent economical losses. Previous surveys indicated the green tides were initiated in the coastal water of southern Jiangsu province where Porphyra farming was intensively conducted; however, the main ‘seed source’ of floating green algae is still under debate. Ulva prolifera was confirmed to be the major causative species of green tides. The multiple sympatric ulvoid species in the natural environment has complicated species identification in both field surveys and laboratory studies due to their morphological plasticity. Thus, we developed a genetic identification key based on restriction fragment length polymorphism (RFLP) analysis of the ITS nuclear marker to discriminate the common Ulva and Blidingia species in the Yellow Sea. Ten genetic lineages (1 in Blidingia, 9 in Ulva) were detected along the coast of China through phylogenetic analysis of ITS sequences. They can be separated by virtual restriction digestion using the four selected restriction enzymes (BspT107 I, EcoO109 I, Hin1 I and VpaK11B I). With additional PCR amplification of the 5S spacer region, we were able to discriminate U. prolifera from Ulva linza. Using this genetic key, we screened macroalgal samples collected from the coast of the Yellow Sea, and the results indicated 6 common lineages (U. prolifera, U. linza, Ulva compressa, Ulva pertusa, Clade 6 and Blidingia sp.) in this region, which could be explicitly distinguished by a single enzyme (BspT107 I) coupled with 5S spacer polymorphism. U. prolifera was confirmed to be present on the Porphyra aquaculture rafts with seasonal variation in the species composition. This genetic key will facilitate our long-term field surveys to investigate the origin of the floating U. prolifera and furthermore to explore its bloom dynamics along the coast of the Yellow Sea. It also provided a framework for the future inclusion of more Ulva species, which will expand the usage of this key.     

Freshwater population of Ulva flexuosa (Ulvaceae,Chlorophyta) as a food source for great pond snail: Lymnaea stagnalis (Mollusca,Lymnaeidae)

Cosmopolitan species of the genus Ulva (Ulvaceae; Chlorophyta) that populate the littoral zone of marine habitats constitute a staple diet for a variety of organisms, particularly snails, shellfish, polychaetes, and birds. Occurrence of Ulva species (e.g., U. flexuosa and U . prolifera) has also been observed in freshwater inland ecosystems that have no contact with saline water. However, the influence of the development of macroalgal mats of Ulva on indigenous organisms in limnic ecosystems has not been established. This study investigates the trophic relationships between Ulva flexuosa and one species of snail from freshwater habitats in central Europe. During the summer, the great pond snail (Lymnaea stagnalis) consumed Ulva as a source of nutrition even when other algae and plants were available. Lymnaea stagnalis consumed an average of 100 mg of Ulva thalli per day. This level of biomass exceeded the consumption of an alternative food source, the shoots of Elodea canadensis. Ulva thalli are more actively consumed by great pond snails than Elodea shoots, and this is expressed in terms of the differences of biomass consumption. It was also observed that the interior of the monostromatic tubular thalli of Ulva flexuosa serves as a protective shelter for juvenile great pond snails.     

Diluted seawater promoted the green tide of Ulva prolifera (Chlorophyta,Ulvales)

Ulva prolifera (Müller) J. Agardh is the main causative species of the 2008 Yellow Sea green tide incident. We investigated the influences of diluted seawater on the vegetative growth and reproductive cell formation of the alga. The thalli that were cultivated under low salinities (10‰ and 20‰) and low pH values (pH around 7.0) showed obvious and steady biomass gain, while those cultivated under high salinities (40‰) and relatively higher pH conditions (pH around 8.0) manifested significant biomass loss. The trend was, however, completely the opposite for reproductive cell formation and there were indications that enrichment, over very wide concentration ranges of both nitrogen and phosphate, could significantly promote vegetative growth. Results also indicated that relatively low salinity and low pH regimes boosted vegetative growth but were unfavorable for reproductive cell formation and vice versa. Based on these results, the possible origin and development mechanisms of the green tide event are discussed. Eutrophication in the Changjiang River estuary and adjacent sea areas, as well as extremely high freshwater inflows before, and during, the flood of 2007 – due to the full operation of large‐scale water facilities in the area – may have both played an important role in the formation and development of the green tide event.     

Molecular assessment of Ulva (Ulvales,Chlorophyta) diversity in Vietnam including the new species U. vietnamensis

Species diversity of Ulva in Vietnam was investigated using three commonly used genetic markers, the nuclear encoded rDNA ITS region and the plastid encoded rbcL and tufA genes. Single locus species delimitation methods, complemented with morphological and ecological information resulted in the delimitation of 19 species. This diversity is largely incongruent with the traditional understanding of Ulva diversity in Vietnam. Only four species identified in this study, U. lactuca, U. reticulata, U. spinulosa, and U. flexuosa, have been previously reported, and seven species, U. ohnoi, U. tepida, U. chaugulii, U. kraftiorum, U. meridionalis, U. limnetica, and U. aragoënsis, are recorded for the first time from Vietnam. Seven genetic clusters could not be associated with species names with certainty. A new species, U. vietnamensis, is described from marine to brackish coastal areas from southern Vietnam based on its morphological and molecular distinctiveness from the currently known Ulva species. A comparison with recent molecular-based studies of Ulva diversity showed that species composition in Vietnam is similar to that of adjacent countries, including Japan, China, as well as Australia. Our study emphasizes the importance of molecular data in the assessment of Ulva diversity, and indicates that a lot of diversity may still remain to be discovered, especially in tropical regions.     

The osmorespiratory compromise in sculpins: impaired gas exchange is associated with freshwater tolerance

We acclimated two species of sculpin, the freshwater prickly sculpin (Cottus asper) and the closely related marine Pacific staghorn sculpin (Leptocottus armatus) to freshwater ( approximately 0 g/L), brackish water (15 g/L), and seawater (30 g/L) for at least 4 wk and examined the relationships between respiration, ion regulation, gill morphology, and freshwater tolerance. The prickly sculpin successfully acclimated to all three salinities and did not experience appreciable changes in plasma osmolality, [Cl-], or mortality. Gill Na+/K+-ATPase activity was lowest in prickly sculpins acclimated to freshwater, their native salinity, and increased during acclimation to seawater. Furthermore, prickly sculpins acclimated to freshwater had a 30% higher P(crit) than fish acclimated to brackish water or seawater; P(crit) is the environmental P(O2) below which an animal can no longer maintain a routine (.-)M(O2), and an increase in P(crit) represents a compromise of respiratory gas exchange. The higher P(crit) observed in prickly sculpins acclimated to freshwater is likely a consequence of their having small, relatively thick gills that increase in thickness (by approximately 1 microm) during freshwater exposure. In contrast, the marine Pacific staghorn sculpin successfully acclimated to brackish water and seawater, but high mortality (25%) was observed after 3 wk of exposure to freshwater. Pacific staghorn sculpins exposed to freshwater suffered significant, 15%-20%, reductions in plasma osmolality and [Cl-], and these losses in plasma ions resulted in a 1.4-fold increase in gill Na+/K+-ATPase activity. Pacific staghorn sculpins have large, thin gills that are not modified in response to salinity acclimation, and as a result, these animals show no respiratory compromise during freshwater acclimation, as evidenced by the lack of change in P(crit), but show significant ion regulatory disturbance. Overall, this study suggests that gill thickening and the resulting respiratory compromise are necessary for freshwater tolerance in sculpins.     

Operculum closing as a defence against predatory leeches in four British freshwater prosobranch snails

The defence reaction of operculum closing in response to the presence of the molluscivorous leech Glossiphonia complanata (L.) and the non-molluscivorous Erpobdella octoculata (L.) was studied in four species of freshwater prosobranch gastropod. Bithynia tentaculata (L.) and Valvata piscinalis (Müller) can distinguish between the leeches, reacting only to G. complanata. V. piscinalis is capable of a greater degree of distance chemoreception of the leech ‘scent’. Valvata cristata Müller and Potamopyrgus jenkinsi (Smith) did not react to either leech. V. cristata may not be a potential prey item for G. complanata, while P. jenkinsi is fed on by the leech, but is a relative newcomer to the freshwater fauna. Animal Ecology Research Group, Department of Zoology, University of Oxford Commonwealth Forestry Institute, Department of Plant Sciences, University of Oxford     

Free-floating Ulva in the southwest Netherlands: species or morphotypes? a morphological,molecular and ecological comparison

Free-floating Ulva L. biomass in the eutrophic brackish ‘Veerse Meer’ lagoon (southwest Netherlands) consists of four morphologically identified species: U. curvata (Kützing) De Toni, U. lactuca L., U. rigida C. Agardh and U. scandinavica Bliding. U. curvata could be recognized easily because of the characteristic central cavity in the holdfast of the attached plants, the arrangement of cells in rows and the single pyrenoid in each cell. U. rigida was distinguished by the thick thallus and the large number of pyrenoids. The Veerse Meer isolate, however, was slightly different from the isolate from the Oosterschelde estuary (the Netherlands). U. lactuca and U. scandinavica showed a high degree of overlap in thallus thickness and cell size, but U. scandinavica usually possessed more pyrenoids. However, doubts have frequently been expressed about the use of some morphological characters in Ulva taxonomy. To determine the validity of such characters in the identification of Ulva species, the morphological variation within and between morphological species was recorded and a molecular data set generated. To detect possible ecophysiological differences between species, optimum temperatures and salinities for growth were determined experimentally. The sequences of the nuclear ribosomal DNA internal transcribed spacer 2 (ITS2) and flanking regions of U. lactuca, U. rigida and U. scandinavica from the Veerse Meer were all identical, but differed from that of U. rigida from the Oosterschelde estuary. Ulva species from the Veerse Meer were most closely related to U. armoricana and U. rigida from Brittany (2.9% and 3.5% divergence respectively); the difference between U. rigida from the Veerse Meer and from the Oosterschelde estuary was 7.5%. Rooted trees, based on a comparison of these sequences with sequences of other Ulva and Enteromorpha species obtained from the literature, using Monostroma arcticum as outgroup, suggested that Ulva is paraphyletic with respect to Enteromorpha. The optimum temperature for growth of U. curvata was 25?°C; for all other species it was 10?°C. The optimum salinity for growth was 30?°C for all isolates. It is concluded that U. lactuca, U rigida and U. scandinavica from the Veerse Meer are all members of one highly polymorphic species.     

The salinity preference of members of the genus Macroplea (Coleoptera, Chrysomelidae, Donaciinae), fully aquatic leaf beetles that occur in brackish water

Reed beetles (Donaciinae) of the genus Macroplea Samouelle, 1819 live permanently submerged. Literature indicates that Macroplea mutica occurs in brackish water, whereas Macroplea appendiculata is restricted to freshwater. The salinity preference of these two species was tested in a linear and a circular device that offered a continuous salinity gradient. The distribution of animals in the devices was monitored over at least 3 h in each of the 21 experiments. Both species preferred freshwater (salinity 0) over brackish water (salinity 10). In particular, this holds true for specimens collected in brackish water. Likewise, immediate reactions could be observed when during such experiments the direction of the gradient was reversed. While M. mutica can be regarded as a truly marine insect, this marine environment does not strictly reflect its fundamental niche with respect to salinity preference. This is in line with accumulating evidence that M. mutica can be found in freshwater habitats (and M. appendiculata in brackish water). This indicates that the species’ distribution might be influenced by other factors like host plant preference or dispersal mechanisms. It is discussed if—in spite of similar fundamental niches—differences in salinity tolerance (and hence performance in brackish water) may have contributed to speciation in the genus Macroplea.     

Novel distribution pattern between coexisting sexual and obligate asexual variants of the true estuarine macroalga Ulva prolifera

Where sexual and asexual forms coexist within a species, the asexuals are often found to be prevalent in marginal habitats. This asexual distribution pattern has received evolutionary attention linked to the paradox of sex. In many marine species, there is a distributional trend of asexual modes being more common in lower salinity waters regarded as the ecogeographic marginal, being explained by negative effects of low salinities on sexual reproductive success. However, the distribution pattern of estuarine species recently adapted to low salinity waters has remained unknown. The brackish macroalga Ulva prolifera being a major benthic component of estuarine ecosystems includes a sexual variant and obligate asexual variants by means of motile spores. We examined the sexual–asexual distribution pattern of this alga along a salinity gradient in river estuaries. Surprisingly, opposite to the distributional trend of marine organisms, the results clearly showed the persistent predominance of sexuals in the lower salinity reaches than the asexuals. In addition, a frequent alternating of dioecious gametophytes and sporophytes in the sexual population was observed, suggesting the sexual reproductive process would be robustly performed in the low salinity waters. Considering U. prolifera had evolved from the ancestral marine species to become a true estuarine species of which the core habitat is the low salinity reaches, in a broad sense its sexual–asexual distribution pattern would be involved in asexual marginal occupations of the species range previously reported in other organisms. Based on the frozen niche variation model, we can give a concise explanation for the evolutionary process of this pattern; multiple asexuals with frozen genotypic variation had arisen from sexual ancestors undergoing low salinity adaptation and share the estuarine habitat with the sexuals at present.     

Photosynthetic activity and proteomic analysis highlights the utilization of atmospheric CO2 by Ulva prolifera (Chlorophyta) for rapid growth

Free‐floating Ulva prolifera is one of the causative species of green tides. When green tides occur, massive mats of floating U. prolifera thalli accumulate rapidly in surface waters with daily growth rates as high as 56%. The upper thalli of the mats experience environmental changes such as the change in carbon source, high salinity, and desiccation. In this study, the photosynthetic performances of PSI and PSII in U. prolifera thalli exposed to different atmospheric carbon dioxide (CO₂) levels were measured. Changes in photosynthesis within salinity treatments and dehydration under different CO₂ concentrations were also analyzed. The results showed that PSII activity was enhanced as CO₂ increased, suggesting that CO₂ assimilation was enhanced and U. prolifera thalli can utilize CO₂ in the atmosphere directly, even when under moderate stress. In addition, changes in the proteome of U. prolifera in response to salt stress were investigated. Stress‐tolerance proteins appeared to have an important role in the response to salinity stress, whereas the abundance of proteins related to metabolism showed no significant change under low salinity treatments. These findings may be one of the main reasons for the extremely high growth rate of free‐floating U. prolifera when green tides occur.     

Larval salinity tolerance of the South American salt-marsh crab, Neohelice (Chasmagnathus) granulata: physiological constraints to estuarine retention, export and reimmigration

The semiterrestrial crab Neohelice (=Chasmagnathus) granulata (Dana 1851) is a predominant species in brackish salt marshes, mangroves and estuaries. Its larvae are exported towards coastal marine waters. In order to estimate the limits of salinity tolerance constraining larval retention in estuarine habitats, we exposed in laboratory experiments freshly hatched zoeae to six different salinities (5–32‰). At 5‰, the larvae survived for a maximum of 2 weeks, reaching only exceptionally the second zoeal stage, while 38% survived to the megalopa stage at 10‰. Shortest development and negligible mortality occurred at all higher salt concentrations. These observations show that the larvae of N. granulata can tolerate a retention in the mesohaline reaches of estuaries, with a lower limit of ca. 10–15‰. Maximum survival at 25‰ suggests that polyhaline conditions rather than an export to oceanic waters are optimal for successful larval development of this species. In another experiment, we tested the capability of the last zoeal stage (IV) for reimmigration from coastal marine into brackish waters. Stepwise reductions of salinity during this stage allowed for moulting to the megalopa at 4–10‰. Although survival was at these conditions reduced and development delayed, these results suggest that already the zoea-IV stage is able to initiate the reimmigration into estuaries. After further salinity reduction, megalopae survived in this experiment for up to >3 weeks in freshwater, without moulting to juvenile crabs. In a similar experiment starting from the megalopa stage, successful metamorphosis occurred at 4–10‰, and juvenile growth continued in freshwater. Although these juvenile crabs showed significantly enhanced mortality and smaller carapace width compared to a seawater control, our results show that the late larval and early juvenile stages of N. granulata are well adapted for successful recruitment in brackish and even limnetic habitats.     

盐分对河口淡水、微咸水沼泽湿地土壤甲烷产生潜力的影响

海平面上升导致河口区盐水入侵现象日益明显,深刻影响着河口潮汐淡水、微咸水湿地生物地球化学循环。采集闽江河口区淡水、微咸水短叶茳芏潮汐沼泽湿地表层土样,室内添加盐度为5,10,15,21 g/L的人造海水、Na Cl溶液及盐度为0的去离子水,通过室内泥浆厌氧培养试验,对比研究海水和Na Cl溶液对淡水、微咸水沼泽湿地土壤甲烷产生潜力的影响。与对照相比,1—12 d培养期内4个盐度的海水处理均显著抑制河口淡水、微咸水沼泽湿地甲烷产生潜力,抑制率在93%以上,盐度10—21 g/L的3个海水处理对于河口淡水、微咸水沼泽湿地甲烷产生潜力的抑制效应无显著差异。Na Cl溶液只有在盐度达到15和21 g/L才显著抑制淡水、微咸水沼泽湿地甲烷产生潜力,且抑制率最多为80.9%,盐度为5、10 g/L的Na Cl溶液对淡水、微咸水沼泽湿地甲烷产生潜力的抑制作用不显著,抑制率多小于30%。伴随着盐水入侵而发生的硫酸盐还原作用及离子胁迫作用对河口淡水、微咸水沼泽湿地甲烷产生具有显著的抑制效应。     

BACTERIA THAT INDUCE MORPHOGENESIS IN ULVA PERTUSA (CHLOROPHYTA) GROWN UNDER AXENIC CONDITIONS1

Marine foliaceous green macroalgae such as Ulva lose their typical morphology when cultured aseptically in defined synthetic media. However, after reinfection by certain marine bacteria (isolated from unialgal cultures of Ulva pertusa Kjellman), the organisms regain their typical foliaceous or tubular morphology. To investigate the morphogenesis (MG) induced in U. pertusa by bacteria, we isolated and identified bacteria with MG activity on U. pertusa and studied the distribution of such bacteria in seawater and on various marine macroalgae. We isolated 1555 bacterial strains from 18 species of marine macroalgae (six Chlorophyta, five Phaeophyta, and seven Rhodophyta), from seawater and from sediment collected at the beach at Omaezaki, Shizuoka Prefecture; Japan. Of these, 676 bacterial strains (43.5%) showed MG activity. They were classified into six bacterial groups, Flavobacterium, Vibrio, Pseudomonas, Deleya, Escherichia, and gram-positive cocci. These bacteria were ubiquitous among the samples and were not specific to U. pertusa. Several plant growth regulators had no MG activity. Filter-sterilized supernatants of culture media of MG-active bacteria strains did not induce MG. Cocultivation of Ulva with active bacterial strains is so far the only way to induce the MG effect, which suggests that for MG direct contact between Ulva and the bacterial strain is necessary.