Sexual systems in scleractinian corals: an unusual pattern in the reef-building species Diploastrea heliopora

The sexual system in corals refers to the spatial and temporal pattern of sexual function within an individual coral polyp, colony or population. Although information on sexual systems now exists for over 400 scleractinian species, data are still lacking for some important reef-building taxa. The vast majority of scleractinians are either simultaneous hermaphrodites or gonochoric with other sexual systems rarely occurring. Diploastrea heliopora is one of the most ubiquitous and easily recognised reef-building species in the Indo-West Pacific; however, surprisingly little is known about its reproductive biology. The aim of the present study was to examine the reproductive biology of D. heliopora colonies on chronically impacted, equatorial reefs south of Singapore. Here we show that in Singapore, D. heliopora is a broadcast spawner with predominantly gonochoric polyps. Colonies, however, contained male, female and a low proportion of cosexual polyps during the 14-month sampling period. The most plausible explanation for this is that polyps switch sexes with oogenic and spermatogenic cycles occasionally overlapping. This leads to colony level alternation of sex function within and between breeding seasons. While this sexual system is atypical for scleractinians, it supports molecular evidence that D. heliopora is phylogenetically distinct from species formerly in the family Faviidae.     

Antioxidant and photoprotective defenses in response to gradual water stress under low and high irradiance in two Malvaceae tree species used for tropical forest restoration

Key message

This study provides a mechanistic basis of the tolerance to light and water stress in two tropical tree species of the Malvaceae used for the reforestation in the tropics. G. ulmifolia is more sensitive to high irradiance than C. speciosa . Thus, C. speciosa is more appropriate for the reforestation of degraded areas with potential excess light or water deficit.

Abstract

Reforestation programs in tropics are necessary to mitigate the impacts of climate change, the preservation of biodiversity and for recovery of degraded areas. However, little is known about the abiotic stress tolerance of the major tropical tree species used in reforestation. This study was aimed at evaluating antioxidant defenses and photoprotective pigments against excess light and water deficit in Guazuma ulmifolia Lam. and Ceiba speciosa (A.St.-Hil) Ravenna. Plantlets of 6-month-old were exposed to progressive water deficit by withholding water under high and low irradiance growth conditions and then re-irrigated. G. ulmifolia leaves under high irradiance showed increased photo-oxidative damage, as indicated by a decreased Fv/Fm ratio and 5?12-fold increases in MDA levels, which correlated with a 14-fold increase in SOD activity, a 90 % decrease in POD activity and a 3?4-fold increases in hydrogen peroxide levels. Water deficit combined with high irradiance caused stronger chronic photoinhibition, indicating a synergistic effect that cannot be counterbalanced by antioxidant or photo-protective mechanisms. In contrast, to avoid photo-oxidative damage, C. speciosa enhanced SOD activity, maintained POD and CAT activities, and increased xanthophyll cycle pool size and DPS and β-carotene accumulation. These results indicate that G. ulmifolia is more sensitive to high irradiance than C. speciosa at early stages of development. Thus, C. speciosa is more appropriate for the reforestation of degraded areas with potential excess light or water deficit. In conclusion, this study provides a mechanistic basis of the tolerance abiotic stress in two tropical trees, thereby contributing for management strategies in reforestation projects.     

Effects of Oil-Contaminated Sediments on Submerged Vegetation: An Experimental Assessment of Ruppia maritima

Oil spills threaten the productivity of ecosystems through the degradation of coastal flora and the ecosystem services these plants provide. While lab and field investigations have quantified the response of numerous species of emergent vegetation to oil, the effects on submerged vegetation remain uncertain. Here, we discuss the implications of oil exposure for Ruppia maritima, one of the most common species of submerged vegetation found in the region affected by the recent Deepwater Horizon oil spill. We grew R. maritima in a range of manipulated sediment oil concentrations: 0, 0.26, 0.53, and 1.05 mL oil /L tank volume, and tracked changes in growth (wet weight and shoot density/length), reproductive activity (inflorescence and seed production), root characteristics (mass, length, diameter, and area), and uprooting force of plants. While no statistical differences were detected in growth, plants exhibited significant changes to reproductive output, root morphology, and uprooting force. We found significant reductions in inflorescences and fruiting bodies at higher oil concentrations. In addition, the roots growing in the high oil were shorter and wider. Plants in medium and high oil required less force to uproot. A second experiment was performed to separate the effects of root morphology and oiled sediment properties and indicated that there were also changes to sediment cohesion that contributed to a reduction in uprooting forces in medium and high oil. Given the importance of sexual reproduction for these plants, oil contamination may have substantial population-level effects. Moreover, areas containing buried oil may be more susceptible to high energy storm events due to the reduction in uprooting force of foundation species such as R. maritima.     

Palytoxin found in Palythoa sp. zoanthids (Anthozoa, Hexacorallia) sold in the home aquarium trade

Zoanthids (Anthozoa, Hexacorallia) are colonial anemones that contain one of the deadliest toxins ever discovered, palytoxin (LD₅₀ in mice 300 ng/kg), but it is generally believed that highly toxic species are not sold in the home aquarium trade. We previously showed that an unintentionally introduced zoanthid in a home aquarium contained high concentrations of palytoxin and was likely responsible for a severe respiratory reaction when an individual attempted to eliminate the contaminant colonies using boiling water. To assess the availability and potential exposure of palytoxin to marine aquarium hobbyists, we analyzed zoanthid samples collected from local aquarium stores for palytoxin using liquid chromatography and high resolution mass spectrometry and attempted to identify the specimens through genetic analysis of 16S and cytochrome c oxidase 1 (COI) markers. We found four specimens of the same apparent species of zoanthid, that we described previously to be responsible for a severe respiratory reaction in a home aquarium, to be available in three aquarium stores in the Washington D.C. area. We found all of these specimens (n = 4) to be highly toxic with palytoxin or palytoxin-like compounds (range 0.5–3.5 mg crude toxin/g zoanthid). One of the most potent non-protein compounds ever discovered is present in dangerous quantities in a select species of zoanthid commonly sold in the home aquarium trade.     

A safe alternative to invasive Caulerpa taxifolia (Chlorophtya)? Assessing aquarium-release invasion potential of aquarium strains of the macroalgal genus Chaetomorpha (Chlorophyta)

Aquarium releases threaten the ecological integrity of aquatic systems by introducing non-native species. Following aquarium-release invasions by Caulerpa taxifolia, other genera of marine macroalgae were promoted by aquarists as alternatives for aquarium hobbyist use. The most popular, Chaetomorpha, was named a preferable alternative to invasive Caulerpa with desirable characteristics including rapid nutrient uptake, broad environmental tolerances and ease of acquisition. As these same characteristics are also associated with invasion success, we assessed the risk posed if aquarium release activities extend to Chaetomorpha. Here we address the propensity for vegetative fragmentation, a primary contributor to the breadth of the Caulerpa invasions, as a potential invasion risk for Chaetomorpha. We monitored fragment generation of 10 purchases of Chaetomorpha from aquarium hobby retailers and tested viability of fragments 0.5–10 mm in length at 5, 22 and 30 °C. We found that Chaetomorpha can survive from fragments as small as 0.5 mm (one live cell). Abundance of viable fragments generated during shipping ranged from 28 to 6,266 per purchase. In 9 of 10 trials, survival was independent of starting length, indicating that small size will not limit potential establishment. Fragments in these purchases had low survival at 5°, but one purchase showed high survival at all temperatures. With high survivorship of small fragments and large numbers of fragments generated, we caution that aquarium strains of Chaetomorpha may pose a threat if released into natural waterways and encourage educational outreach strategies which focus on changing releasing behaviors rather than promoting “safe” alternatives to invasive ornamentals.     

Molecular genetic identification and phylogeny of Daphnia species (Crustacea, Cladocera) from water bodies of the Lake Chany basin

The data on the molecular genetic identification of Daphnia species from the water bodies of the Lake Chany basin are presented. Phylogenetic relationships between these species have been established. The fragments of the mitochondrial DNA genes were used as genetic markers. According to the data obtained, the water bodies examined were inhabited by five Daphnia species, including Daphnia (Daphnia) galeata Sars, D. (D.) longispina O. F. Müller, D. (D.) curvirostris Eylmann, D. (D.) pulex Leydig, and D. (Ctenodaphnia) magna Straus. In addition, longispina a-like individuals that form a separate phylogenetic lineage was identified.     

Taxonomy and species boundaries in the coral genus Favia Milne Edwards and Haime, 1857 (Cnidaria: Scleractinia) from Thailand revealed by morphological and genetic data

While Faviidae is a widely and uniformly distributed coral family throughout the Indo-Pacific, the extensive phenotypic plasticity of colony surface and corallite features often confounds the use of macromorphological characters in species identification, and contributes to conflict between traditional classification and molecular analyses of the group. Recent advances in morphological and molecular techniques now provide a suite of methods to re-address coral taxonomy in complex groups, such as that represented by the Faviidae. This study combines morphologic measurements including “3D coordinates landmarks” data with phylogenetic assessments of nuclear (ITS) and mitochondrial (COI-trnM) DNA to assess species boundaries in nine species of Faviidae with para-septothecal walls from Thailand. Strong concordance was found between morphological features and a priori groupings based on both morphospecies and genetically defined groups (ITS and COI-trnM). Favia truncatus was the most well-defined species based on morphological analyses, and it was also shown to be monophyletic using phylogenetic analyses. Besides F. truncatus, the only other species that was found to be monophyletic in analyses of both genes was F. cf. helianthoides, but its skeletal morphology overlapped with the F. favus species complex (comprised of F. favus, F. speciosa, F. matthaii and F. rotumana). Although not genetically monophyletic, the F. favus species complex and F. pallida were fairly well delineated morphologically. Morphospecies within the F. favus species complex are therefore possibly a result of genetic drift and/or stable polymorphisms driven by divergent selection. These results represent a first step toward a taxonomic revision of the Indo-Pacific Favia, which will integrate morphological methods with the study of type material, genetic information, reproductive data, and tests of phenotypic plasticity—given that multiple lines of evidence are needed to resolve ambiguous species and assign species names.     

Characterization of the complete mitochondrial genome of Diploastrea heliopora and phylogeny of the scleractinia species which have group I introns in their COI genes

Mitochondrial genome DNA is a powerful marker for resolving phylogenetic relationships among scleractinian corals. Here, we decode the complete mitochondrial genome of Diploastrea heliopora (Lamarck, 1816) for the first time. The general features are 18 363 bp in length, and conventionally, with 13 protein coding genes, two ribosomal RNAs, and two transfer RNAs. Gene arrangement and distribution are similar to other scleractinian corals. Moreover, the COI gene of D. heliopora is broken up into two parts by a complex group I intron. This intron is 1076 bases in length and contains helical structures (P1-P10, except P2) and four conserved regions (P, Q, R, and S). The mitochondrial genome of D. heliopora has asymmetric base composition (13.03% C, 20.29% G, 25.91% A, and 40.77% for T). Based on concatenated protein coding genes, ML and BI trees show similar phylogenetic relationship: D. heliopora clustered closely with Sclerophyllia maxima and Echinophyllia aspera into the robust branch. The data and conclusion in this study are reference for further phylogenetic studies of corals.     

Metabolic plasticity of the corals Porites lutea and Diploastrea heliopora exposed to large amplitude internal waves

The metabolic plasticity of the two mounding coral species Porites lutea (Milne-Edwards and Haime, 1860) and Diploastrea heliopora (Lamarck, 1816) was investigated in the Similan Islands (Thailand), an offshore Andaman Sea island group subjected to large amplitude internal waves (LAIW). Nutrient concentrations were highly correlated with LAIW intensity and contributed to 3- and 10-fold higher symbiont densities in P. lutea and D. heliopora, respectively, along with elevated pigment concentrations, protein content, host tissue, and symbiont biomass. The comparison of LAIW-exposed and LAIW-sheltered island faces, and LAIW-intense and LAIW-weak years suggests a species-specific metabolic plasticity to LAIW, where D. heliopora benefits more from increased nutrient and organic matter availability than P. lutea. The ubiquitous LAIW in Southeast Asia and beyond may provide so far unexplored clues to coral acclimatization to disturbances on various scales, and hence, a potential key to coral resilience to climate change.     

High diversity of Vibrio spp. associated with different ecological niches in a marine aquaria system and description of Vibrio aquimaris sp. nov

The aim of the study was to characterise the diversity and niche-specific colonization of Vibrio spp. in a marine aquaria system by a cultivation-dependent approach. A total of 53 Vibrio spp. isolates were cultured from different ecological niches in a marine aquarium including microplastic (MP) and sandy sediment particles (12 weeks after added sterile to the system), detritus, and the surrounding aquarium water. Based on the 16S rRNA gene sequence phylogeny and multilocus sequence analysis (MLSA) the isolates were assigned to seven different phylotypes. Six phylotypes were identified by high probability to the species level. The highest phylotype diversity was cultured from detritus and water (six out of seven phylotypes), while only two phylotypes were cultured from MP and sediment particles. Genomic fingerprinting indicated an even higher genetic diversity of Vibrio spp. at the strain (genotype) level. Again, the highest diversity of genotypes was recovered from detritus and water while only few partially particle-type specific genotypes were cultured from MP and sediment particles. Phylotype V-2 formed an independent branch in the MLSA tree and could not be assigned to a described Vibrio species. Isolates of this phylotype showed highest 16S rRNA gene sequence similarity to type strains of Vibrio japonicus (98.5%) and Vibrio caribbeanicus (98.4%). A representative isolate, strain THAF100^T, was characterised by a polyphasic taxonomic approach and Vibrio aquimaris sp. nov., with strain THAF100^T (=DSM 109633^T = LMG 31434^T = CIP 111709^T) as type strain, is proposed as novel species.     

Sexual dimorphism in Diabrotica speciosa and Diabrotica viridula (Coleoptera: Chrysomelidae)

Diabrotica speciosa (Germar) and Diabrotica viridula F. (Coleoptera: Chrysomelidae) are the two most abundant species of the genus in South America, and belong to the fucata and virgifera groups, respectively. Here, we characterize the dimorphism of the setae present on the basitarsi of males and females of these species. Dimorphism was confirmed in both species, and it was related to the presence of adhesive setae exclusively in males, which possess these structures on the basal tarsomeres of the pro- and mesothoracic legs.     

Photomorphogenesis in Sinningia speciosa, cv. Queen Victoria I. Characterization of Phytochrome Control

The morphological development of Sinningia speciosa plants that were exposed to supplementary far red light was very different from that of plants receiving dark nights. After several nights of such irradiation, stems and petioles were elongated, petioles were angulated, leaf blade expansion was inhibited, plants were chlorotic and the accumulation of shoot dry weight was retarded.

Red reversibility of the morphological changes potentiated by far red light indicated control by the phytochrome system. A high P_FR level during the last half of the night inhibited stem elongation and promoted leaf blade expansion, but both of these processes were hardly affected by the P_FR level during the first half of the night. Thus sensitivity to P_FR was cyclic.

The interpretation of our experiments was complicated by quantitative morphological differences resulting from long, as compared to short, far red irradiations.

     

Surveillance of fish species composition using environmental DNA

Prompt and accurate methods for assessing the species composition of given areas are indispensable in addressing the rapid loss of biodiversity. Here, we propose a method for the surveillance of fish species composition in freshwater using environmental DNA as species markers. First, the applicability of the method was demonstrated through aquarium experiments. DNA was extracted from 120?ml aquarium water, and the degenerated primers targeting the fish mitochondrial cytochrome b gene were used for amplification. PCR-amplified fragments were analysed by random cloning, and all species reared in the aquarium were detected. Next, this method was applied to natural freshwater environments. Water samples were collected from three sites in the Yura River, Japan; DNA was concentrated from 2?l of environmental water, and then amplified and cloned. Up to four species of fish were detected by sequencing 47 randomly selected clones from a single water sample. Overall, the results were consistent with previous knowledge of fish habitat utilisation. Using this method, the surveillance of fish species composition can be conducted less laboriously than with traditional methods.     

Contrasting Light Spectra Constrain the Macro and Microstructures of Scleractinian Corals

The morphological plasticity of scleractinian corals can be influenced by numerous factors in their natural environment. However, it is difficult to identify in situ the relative influence of a single biotic or abiotic factor, due to potential interactions between them. Light is considered as a major factor affecting coral skeleton morphology, due to their symbiotic relation with photosynthetic zooxanthellae. Nonetheless, most studies addressing the importance of light on coral morphological plasticity have focused on photosynthetically active radiation (PAR) intensity, with the effect of light spectra remaining largely unknown. The present study evaluated how different light spectra affect the skeleton macro- and microstructures in two coral species (Acropora formosa sensu Veron (2000) and Stylophora pistillata) maintained under controlled laboratory conditions. We tested the effect of three light treatments with the same PAR but with a distinct spectral emission: 1) T5 fluorescent lamps with blue emission; 2) Light Emitting Diodes (LED) with predominantly blue emission; and 3) Light Emitting Plasma (LEP) with full spectra emission. To exclude potential bias generated by genetic variability, the experiment was performed with clonal fragments for both species. After 6 months of experiment, it was possible to detect in coral fragments of both species exposed to different light spectra significant differences in morphometry (e.g., distance among corallites, corallite diameter, and theca thickness), as well as in the organization of their skeleton microstructure. The variability found in the skeleton macro- and microstructures of clonal organisms points to the potential pitfalls associated with the exclusive use of morphometry on coral taxonomy. Moreover, the identification of a single factor influencing the morphology of coral skeletons is relevant for coral aquaculture and can allow the optimization of reef restoration efforts.     

Morphological variation in the polyps of the scleractinian coral Favia speciosa (Dana) around Singapore

A photographic technique was used to examine morphological differences in the living polyps of Favia speciosa sampled from three sites around Singapore. Eight characters were measured, seven of which differed significantly between the three study sites. Sedimentation rates and character size were much higher at the site closest to the mainland than at the two sites further from shore. Land reclamation and dredging contribute to high sediment rates in Singapore waters; these rates decrease with increasing distance from shore. Large polyps close to the main island of Singapore are possibly a plastic, or selected for, response to high levels of sediment.     

Impacts of Sediments on Coral Energetics: Partitioning the Effects of Turbidity and Settling Particles

Sediment loads have long been known to be deleterious to corals, but the effects of turbidity and settling particles have not previously been partitioned. This study provides a novel approach using inert silicon carbide powder to partition and quantify the mechanical effects of sediment settling versus reduced light under a chronically high sedimentary regime on two turbid water corals commonly found in Singapore (Galaxea fascicularis and Goniopora somaliensis). Coral fragments were evenly distributed among three treatments: an open control (30% ambient PAR), a shaded control (15% ambient PAR) and sediment treatment (15% ambient PAR; 26.4 mg cm⁻² day⁻¹). The rate of photosynthesis and respiration, and the dark-adapted quantum yield were measured once a week for four weeks. By week four, the photosynthesis to respiration ratio (P/R ratio) and the photosynthetic yield (F_v/F_m) had fallen by 14% and 3–17% respectively in the shaded control, contrasting with corals exposed to sediments whose P/R ratio and yield had declined by 21% and 18–34% respectively. The differences in rates between the shaded control and the sediment treatment were attributed to the mechanical effects of sediment deposition. The physiological response to sediment stress differed between species with G. fascicularis experiencing a greater decline in the net photosynthetic yield (13%) than G. somaliensis (9.5%), but a smaller increase in the respiration rates (G. fascicularis = 9.9%, G. somaliensis = 14.2%). These different physiological responses were attributed, in part, to coral morphology and highlighted key physiological processes that drive species distribution along high to low turbidity and depositional gradients.     

Effect of Biotic and Abiotic Factors on In Vitro Proliferation, Encystment, and Excystment of Pfiesteria piscicida

Pfiesteria spp. are mixotrophic armored dinoflagellates populating the Atlantic coastal waters of the United States. They have been a focus of intense research due to their reported association with several fish mortality events. We have now used a clonal culture of Pfiesteria piscicida and several new environmental isolates to describe growth characteristics, feeding, and factors contributing to the encystment and germination of the organism in both laboratory and environmental samples. We also discuss applied methods of detection of the different morphological forms of Pfiesteria in environmental samples. In summary, Pfiesteria, when grown with its algal prey, Rhodomonas sp., presents a typical growth curve with lag, exponential, and stationary phases, followed by encystment. The doubling time in exponential phase is about 12 h. The profiles of proliferation under a standard light cycle and in the dark were similar, although the peak cell densities were markedly lower when cells were grown in the dark. The addition of urea, chicken manure, and soil extracts did not enhance Pfiesteria proliferation, but crude unfiltered spent aquarium water did. Under conditions of food deprivation or cold (4°C), Pfiesteria readily formed harvestable cysts that were further analyzed by PCR and scanning electron microscopy. The germination of Pfiesteria cysts in environmental sediment was enhanced by the presence of live fish: dinospores could be detected 13 to 15 days earlier and reached 5- to 10-times-higher peak cell densities with live fish than with artificial seawater or f/2 medium alone. The addition of ammonia, urea, nitrate, phosphate, or surprisingly, spent fish aquarium water had no effect.     

Characterization of fungi from hypersaline environments of solar salterns using morphological and molecular techniques

The Cabo Rojo Solar Salterns located on the southwest coast of Puerto Rico are composed of two main ecosystems (i.e., salt ponds and microbial mats). Even though these locations are characterized by high solar radiation (mean light intensity of 39 mol photons m⁻² d⁻¹) they harbour a diverse microscopic life. We used morphological and molecular techniques to identify a series of halotolerant fungi. A total of 183 isolates and 36 species were cultured in this study. From the water from the salt ponds, 86 isolates of 26 species were cultured. The halotolerant fungi isolated from water were: Cladosporium cladosporioides, nine Aspergillus sp., five Penicillium sp. and the black yeast Hortaea werneckii. A distinctive isolate with a blue mycelium was cultured from the salt ponds, representing a new species of Periconia based on morphology and rDNA analysis. Forty-four isolates from eight species were cultured from the sediments around the salt ponds. Most of the sediment isolates formed only sterile mycelium, while several were Chaetomium globosum. A total of 53 isolates from 16 species were isolated from the three layers of the microbial mats, of which Aspergillus niger was the most frequent isolate. Phospholipid fatty acid profiles generated from the different layers of the microbial mats indicated that the uppermost layers of the mats contained fungal biomarker, 18:2w6. This fatty acid decreased with depth, the highest concentration was observed in the green upper layer and it disappeared in the black bottom anoxic layer. This correlates with the isolation of fungi using the serial dilution technique. This is the first study that documents the presence of fungi in microbial mats.     

Root hydraulic conductance,gas exchange and leaf water potential in seedlings of Pistacia lentiscus L. and Quercus suber L. grown under different fertilization and light regimes

Differences in morphology, biomass allocations and physiological responses were investigated in seedlings of Mastic tree (Pistacia lentiscus L.) and Cork oak (Quercus suber L.) submitted to contrasting fertilization and light regimes during early growth. These species are two evergreen sclerophyllous Mediterranean species frequently used in Mediterranean reforestation programmes. Fertilization was the treatment that affected most of the morphological and physiological variables evaluated in P. lentiscus and Q. suber seedlings. Leaf area and specific leaf area (SLA) were affected by shading treatment in both species, showing higher values in seedlings grown under shade. P. lentiscus seedlings showed a high capacity to modify root morphological variables and root hydraulic conductance (K_R) with the fertilization treatment. In contrast, Q. suber showed low to moderate root system changes with the treatments applied, although the fertilization level affected biomass allocation (i.e., root to shoot ratio) in both species. Under high water demand, P. lentiscus seedlings with high K_R allowed transpiration (E) to increase without increasing the water potential gradient between soil and leaves. In Q. suber, high fertilization induced significant increases in photosynthesis (A), as well as a tendency to increase E with significantly lower leaf water potential (ψ_L).