Diversity of phlorotannin profiles among sargassasacean species affecting variation and abundance of epiphytes

In general, epiphytes have detrimental effects on the growth of their basiphytes due to competition for light and nutrients. Therefore, basiphyte species must expend energy suppressing epiphytes. Some studies suggest that phlorotannins, i.e. brown algal polyphenols, prevent colonization by epiphytes, whereas others question their allelopathic function because there is not necessarily a negative correlation between epiphyte abundance and the phlorotannin content of the basiphyte algae. Various phlorotannin components are found in brown algal species, thus we hypothesized that the antifouling activities of polyphenolic compounds may differ and that the analysis of phlorotannin profiles could be useful for estimating their ecological functions. We surveyed the epiphyte richness in the apical portions of 373 thalli from 15 sargassacean species, demonstrating that the variation and abundance of epiphyte species differed remarkably among the basiphyte species. However, there was a weak negative correlation between the density and total phlorotannin content of the basiphyte algae in only one of the 18 epiphyte species. The interspecific differences in the phlorotannin profile were characterized by quantitative ¹H nuclear magnetic resonance spectroscopy (qNMR), and four major groups were categorized based on cluster and principal component analyses of polyphenolic signals in the qNMR spectra. The epiphyte Neosiphonia harveyi was more abundant on Sargassum hemiphyllum, S. patens and S. piluliferum than on other basiphyte species, and these three species were similar according to the cluster analysis. These results suggest that some phlorotannin components may be more effective for antifouling; thus interspecific differences in the phlorotannin profile could affect the variation and abundance of epiphytes.     

Floating period of Sargassacean thalli estimated by the change in density

The density (fresh weight/volume) of the detached thalli of four sargassacean species (Myagropsis myagroides, Sargassum horneri, S. patens, S. siliquastrum) was measured weekly to determine their floating period. Thalli of the four species were collected with respect to the difference in their reproductive stage, and their floating abilities at different stages were determined. Floating thallus was ballasted with pre-weighed leads until the final addition of a weight caused sinking. The density and floating period of sargassacean thalli were dependent on the species and the specimens’ reproductive stage. The floating period of thalli detached before maturation was estimated to be between 4 and 14 weeks. However, thalli detached during maturation had a floating period of between 1 and 8 weeks, and those detached after maturation sank within 2 weeks. Sargassum horneri had the lowest density and the longest floating period among the four species investigated and this may explain the high abundance of S. horneri in floating seaweeds observed in the Sea of Japan.     

QUANTITATIVE VARIATION IN VULPINIC AND PINASTRIC ACIDS PRODUCED BY TUCKERMANNOPSIS PINASTRI (LICHEN-FORMING ASCOMYCOTINA,PARMELIACEAE)

Tuckermannopsis pinastri is a lichen species found commonly on rocks and tree branches in boreal and alpine habitats in the northern United States. Members of this species produce three yellow-pigmented phenolic compounds: usnic, pinastric, and vulpinic acids. The objective of our study was to quantify the variation in concentration of the latter two of these compounds in relation to substrate factors, chlorophyll content, and thallus size. Using high-performance liquid chromatography, we analyzed 120 thalli of T. pinastri collected randomly from a single large population located at Spruce Knob, West Virginia. Although individuals were sampled from both tree and rock substrates that differed markedly in light intensity, these environmental factors were not correlated with observed variations in vulpinic or pinastric acid concentrations. Instead, compound concentrations were correlated most closely with thallus size, with small rather than large thalli having the highest concentrations of the two compounds. Small thalli did not have higher concentrations of chlorophylls than large thalli, however, which suggests that the rate of production of secondary compounds by the fungus in T. pinastri is independent of algal biomass. Inasmuch as lichen secondary compounds serve a defensive role against microorganisms and herbivores, our results suggest that small, juvenile thalli are better defended than more mature thalli.     

IMPACT OF ENVIRONMENTAL CHANGES ON PRODUCTION OF BROWN ALGAL PHLOROTANNINS

Targett, N. M. Graduate College of Marine Studies, University of Delaware, Lewes, DE 19958 USA Ecological theories such as resource availability theory (e.g. carbon-nutrient balance hypothesis) and growth differentiation balance theory have attempted to provide a theoretical framework in which to balance putative improvements in plant fitness resulting from secondary metabolite production with the metabolic costs of these compounds. However, experimental tests of these theories have yielded mixed results, in part because they have relied upon static measures of phlorotannin concentrations and have incorrectly assumed phlorotannins to be immobile defenses with negligible rates of turnover. To test these theories more effectively, in situ rates of phlorotannin biosynthesis were examined in several species of brown algae as they responded to manipulations of their surrounding environmental conditions (light, nutrients, and simulated herbivory). The rates of phlorotannin synthesis measured for damaged Fucus vesiculosus from Maine (USA) were dramatically in-creased relative to controls. Carbon resources required for this response were supplied via wound-induced increases in photosynthetic rates, rather than changes in patterns of internal resource allocation. The lack of internal resource trade-offs indicates that phlorotannin production may not always generate allocation costs in these plants. Shading decreased the magnitude of the induced response but growth vs. defenses trade-offs were not identified. For Sargassum hystrix var. buxifolium from the Bahamas, rates of phlorotannin synthesis and total phenolic contents were decreased by both shading and nutrient enrichment. Overall, rates of phlorotannin synthesis were often, but not always, correlated with the eventual changes in total phlorotannin concentrations. Where synthesis and accumulations were uncoupled, differential rates of phlorotannin turnover are expected.     

Tropical marine herbivore assimilation of phenolic-rich plants

Phenolics in marine brown algae have been thought to follow a latitudinal gradient with high phenolic species in high latitudes and low phenolic species in low latitudes. However, tropical brown algae from the western Caribbean have been shown to be high in phlorotannin concentration, indicating that latitude alone is not a reasonable predictor of marine plant phenolic concentrations. This study shows that the range of high phenolic phaeophytes is not limited to the western Caribbean but encompasses the western tropical Atlantic, including Bermuda and the Caribbean, where algal phlorotannin concentrations can be as high as 25% dry weight (DW). Assimilation efficiencies (AEs) of phenolic-rich and phenolic-poor plants were examined in three tropical marine herbivores (the parrotfish, Sparisoma radians, and the brachyuran crab, Mithrax sculptus, from Belize and the parrotfish, Sparisoma chrysopterum, from Bermuda). AEs of phenolic-rich food by each of the three herbivore species were uniformly high, suggesting that high plant phenolic concentrations did not affect AEs in these species. This is in contrast to some temperate marine herbivores where phenolic concentrations of 10% DW have been shown to drastically reduce AE. The apparent contradiction is discussed in light of the effects of specific herbivore gut characteristics on successful herbivory of high phenolic brown algae.     

EFFECTS OF UV RADIATION ON GROWTH AND PHLOROTANNINS IN FUCUS GARDNERI (PHAEOPHYCEAE) JUVENILES AND EMBRYOS1

Diminishing levels of atmospheric ozone are increasing UV stress on intertidal algae. Early developmental stages tend to be more susceptible to environmental stresses; however, little research has examined how these stages are protected from UV radiation (UVR). Many brown algae contain high levels of phlorotannins, which are thought to function in screening UVR. In this study, we tested the effects of ambient levels of UV‐B and UV‐A on growth and phlorotannin production in 1‐ to 2‐cm juvenile and microscopic postsettlement embryos of the intertidal alga Fucus gardneri Silva. Algae were grown in four light treatments: 1) ambient light; 2) under cellulose acetate, which lowered light quantity but did not affect light quality; 3) under Mylar^TM, which filtered UV‐B; and 4) under Plexiglas^TM, which blocked UV‐A and UV‐B. Over a 3‐week period, UV‐B inhibited and UV‐A enhanced the growth of F. gardneri embryos, whereas the growth of juveniles was not affected. Phlorotannin concentrations of both embryos and juveniles did not differ in any of the light treatments. Our results suggest that embryos of F. gardneri are susceptible to UV light but develop a tolerance to it as they mature. This tolerance may result from increases in phlorotannin concentrations that occur during maturation; however, phlorotannin production in embryonic or juvenile stages is either not induced by UV light or takes more than 3 weeks to occur.     

氨基酸叶面硒肥对铁皮石斛富硒及多糖含量的影响

研究不同浓度氨基酸叶面硒肥(CK、1200倍液、900倍液和600倍液)对智能温室人工栽培红杆铁皮石斛(Dendrobium officinale)枝条(即鲜条)富硒、抽芽和多糖含量的影响。结果表明,氨基酸硒肥各浓度处理均极显著提高鲜条硒和多糖含量。鲜条硒含量与氨基酸硒肥浓度呈正相关,且各处理间达极显著差异。鲜条多糖含量在氨基酸硒肥浓度较低(CK、1200、900倍液)时呈正相关,并在氨基酸硒肥900倍液时达最高(35.213 g·kg-1),之后呈递减趋势。氨基酸硒肥稀释达900倍时,90%以上鲜条已抽出3片韧叶,表明氨基酸硒肥明显促进鲜条生长。因此,环保型氨基酸硒肥900倍液是铁皮石斛营养积累前期较适宜浓度。     

Responses of growth and phlorotannins in <Emphasis Type="Italic">Fucus vesiculosus</Emphasis> to nutrient enrichment and herbivory

Discharge from anthropogenic sources may modify both macroalgal growth patterns and resource allocation to carbon based secondary compounds, thereby affecting their susceptibility for herbivory. We tested the effect of eutrophication in terms of nutrient enhancement on growth and phlorotannin concentration of Fucus vesiculosus by conducting manipulative experiments in the field and mesocosms. In the field experiment we utilised fish farms as nutrient sources and in the mesocosm-experiment we manipulated ambient nutrient levels and occurrence of the herbivorous isopod Idotea baltica. Vicinity of a fish farm affected neither growth nor the phlorotannin concentration of Fucus but increased the amount of epiphytes growing on Fucus. Other organisms such as epiphytic filamentous algae and periphyton, which are more capable of quickly utilizing excess nutrients, may restrain the direct effects of nutrient enhancement on Fucus. In a manipulative mesocosm experiment, neither nutrient enrichment nor occurrence of herbivores affected phlorotannin concentration implying lack of induced defences, at least in terms of increasing phlorotannin concentration. Feeding of thallus decreased the growth rate of algae, but the number of reproductive organs, receptacles, was not affected by herbivory. The negative effect of herbivory on the amount of apical tips tended to be stronger under nutrient enriched conditions. We conclude that eutrophication processes, in terms of nutrient enrichment, does not have strong direct effect on growth or phlorotannin production of F. vesiculosus. However, there may be important indirect consequences. First, herbivory may be targeted more to apical parts of the thallus under eutrophicated conditions. Second, the result that Fucus growing close to nutrient sources were smaller than those in control areas may reflect differences in mortality schedules of algae between eutrophicated and control areas.     

Microphytoplankton of the Pitchavaram mangals,southeast coast of India: species composition and population density

Species richness of phytoplankton of the Pitchavaram mangals was high. There were 82 species constituted by 67 species of diatoms, 12 species of dinoflagellates and 3 species of bluegreen algae. The diatoms formed the bulk with 72% followed by the dinoflagellates with 15%, in which there were autochthonous (temporary and permanent) and allochthonous forms. Phytoplankton population density exhibited a wide seasonal fluctuation with the minimum during monsoon and the maximum during summer, suggesting the possible differential influence of various environmental factors.     

Changes in morphological plasticity of Ulva prolifera under different environmental conditions: A laboratory experiment

The large-scale green tides, consisting mainly of Ulva prolifera, have invaded the coastal zones of western Yellow Sea each year since 2008, resulting in tremendous impacts on the local environment and economy. A large number of studies have been conducted to investigate the physiological traits of U. prolifera to explain its dominance in the green tides. However, little has been reported regarding the response of U. prolifera to changing environmental factors via morphological variation. In our experiments, we found remarkable morphological acclimation of U. prolifera to various temperature (20 and 25 °C) and salinity (10, 20, and 30) conditions. U. prolifera had more, but shorter branches when they were cultured at lower temperature and salinity conditions. To investigate the significance of these morphological variations in its acclimation to changes of environmental factors, physiological and biochemical traits of U. prolifera grown under different conditions were measured. Higher temperature increased the relative growth rate while salinity did not affect it. On the other hand, higher temperature did not enhance the net photosynthetic rate whilst lower salinity did. The increased net photosynthetic rate at lower salinity conditions could be attributed to more photosynthetic pigments—chlorophyll a, chlorophyll b, and carotenoids—in thalli due to there being more branches at lower salinity conditions. Increased numbers of branches and thus an increased intensity of thalli may be helpful to protect thalli from increased osmotic pressure caused by lower salinity, but it led to more shading. In order to capture enough light when being shaded, thalli of U. prolifera synthesized more photosynthetic pigments at lower salinity levels. In addition, higher temperature increased nitrate reductase activity and soluble protein content but variations in salinity did not impose any effect on them. Our results demonstrate conclusively that U. prolifera can acclimatize in the laboratory to the changes of environmental factors (salinity and temperature) by morphology-driven physiological and biochemical variation. We suggest that the morphological plasticity of U. prolifera may be an important factor for it to outcompete other algal species in a changing ocean.     

Photobiont association and genetic diversity of the optionally lichenized fungus Schizoxylon albescens

The fungus Schizoxylon albescens occurs both as lichen and as saprobe. Lichenized colonies grow on the bark of Populus tremula; saprotrophic morphs grow on dead Populus branches. We wanted to (1) test whether lichenized and saprotrophic S. albescens are genetically distinct, (2) investigate photobiont association and diversity, (3) investigate the interactions between fungi and algae that occur during co-cultivation and (4) test whether Schizoxylon shows algal selectivity during lichenization. Fungal and algal genetic diversity were investigated for three markers. Algae from lichenized thalli were isolated in axenic cultures, and isolate sequence diversity was compared with algae amplified directly from thallus fragments. Co-culture experiments of fungi and algae were performed to study the morphological interaction patterns. Two distinct phylogenetic units are revealed in S. albescens, which are interpreted as phenotypically cryptic species. The algae are related to Coccomyxa and Pseudococcomyxa, and form two distinct sister clades separating samples isolated in cultures from those amplified directly from thallus fragments, indicating that more easily cultured strains of algae are not necessarily major components of the lichens. Schizoxylon albescens interacts with isolated algal strains, similar to fungal-Coccomyxa symbioses in nature. As the system is maintained without difficulty in culture, it can potentially be an easily controlled lichen symbiosis study system under laboratory conditions.     

Abiotic factors affecting the development of <Emphasis Type="Italic">Ulva</Emphasis> sp. (Ulvophyceae; Chlorophyta) in freshwater ecosystems

The influence of physicochemical factors on the development of Ulva species with distromatic tubular morphology was studied in three streams located in Poznan, Poland. The study evaluated key environmental factors that may influence the colonisation and growth of Ulva populations in freshwater systems. In total, nine environmental parameters were included: temperature, water depth, pH, oxygen (O₂), ammonium (NH₄ ⁺), nitrate (NO₃ ⁻), phosphate (PO₄ ³⁻), sodium chloride (NaCl) and total iron (Fe). Morphometric features of thalli (length and width, percentage of furcated and young thalli) and surface area of free-floating mats formed by the freshwater populations of Ulva were compared at all sites. Principal components analysis indicated the most important factors influencing Ulva development were sodium chloride concentrations and water depth. Two other key chemical factors affecting the freshwater form of Ulva were phosphate and nitrite concentrations. High concentrations of sodium chloride inhibited the development of Ulva, leading to a lower number of thalli in the Ulva mats. At the sites with stable and deeper water, the surface area of the mats was larger. Both phosphate and nitrite concentrations were positively correlated with an increase in the number of thalli in the mats and the thalli length.     

Defense mechanisms of sargassacean species against the epiphytic red alga Neosiphonia harveyi

Flora diversity and abundance of epiphytes are specific to their basiphyte species and may relate to variations in the defensive abilities of basiphytes. Thus, investigating the interactions between epiphytes and basiphytes is useful for a better understanding of the biological impact of epiphytism and the survival strategies of basiphytes. We examined the epiphyte density on five sargassacean species at six locations between two study sites, which showed that the epiphytic red alga Neosiphonia harveyi was remarkably less abundant on Sargassum siliquastrum at all locations. To assess its defense mechanism against N. harveyi, we performed bioassays of phlorotannins, which are considered effective in deterring fouling, by culturing sargassacean blades with N. harveyi carpospores and observed the process by which sargassacean blades remove epiphytes. When the carpospores were incubated with various concentrations of dissolved phlorotannins, settlement and germination were inhibited only at the highest concentrations (>0.1 g · L^?1), and this effect did not significantly differ among the five sargassacean species. When the carpospores were combined with blades from the five species, many of the spores attached and germinated on every blade. Because N. harveyi penetrated rhizoids into basiphyte tissues, cuticle peeling observed in all five sargassacean species could not remove this epiphyte after germination. However, in S. siliquastrum, the blade tissues around the germlings became swollen and disintegrative, and were removed together with the germlings. The spores normally grew on the dead blades, suggesting that the tissue degradation of S. siliquastrum is triggered by the infection of N. harveyi.     

Geographical divergence in host use ability of a marine herbivore in alga–grazer interaction

When the selective environment differs geographically, local herbivore populations may diverge in their host use ability and adapt locally to exploit the sympatric host population. We tested whether populations of the marine generalist herbivore Idotea baltica have diverged in host us ability and whether they locally adapted to exploit the sympatric population of their main host species, the bladderwrack Fucus vesiculosus. We fed isopods from three local populations reciprocally with the sympatric and two allopatric populations of the host. The bladderwrack populations varied in their quality as food for isopods suggesting variation in the selective environment. The ability to exploit the main host showed considerable divergence among the isopod populations. There was no significant interaction between host and isopod origin, indicating that the patterns observed in the reciprocal feeding experiment could be explained by differences in overall suitability of the hosts and differences in overall performance of the isopod populations. Isopod population that was sympatric to a bladderwrack population with low phlorotannin content showed high performance on the algae from the sympatric but low performance on the algae from the two allopatric populations. Performance of isopods, especially in this population, decreased quickly with the increasing phlorotannin content of food algae. We therefore hypothesize that the isopods adapted to a low phlorotannin content were unable to utilize high-phlorotannin algae efficiently. Isopod populations sympatric to the high-phlorotannin bladderwrack populations may be generally better adapted to deal with phlorotannins, being thereby able to utilize a range of bladderwrack populations.     

Deciphering the developmental plasticity of walnut saplings in relation to climatic factors and light environment

Developmental plasticity, the acclimation of plants to their local environment, is known to be crucial for the fitness of perennial organisms such as trees. However, deciphering the many possible developmental and environmental influences involved in such plasticity in natural conditions requires dedicated statistical models integrating developmental phases, environmental factors, and interindividual heterogeneity. These models should be able to analyse retrospective data (number of leaves or length of annual shoots along the main stem in the present case). In this study Markov switching linear mixed models were applied to the analysis of the developmental plasticity of walnut saplings during the establishment phase in a mixed Mediterranean forest. In the Markov switching linear mixed models estimated from walnut data sets, the underlying Markov chain represents both the succession and lengths of growth phases, while the linear mixed models represent both the influence of climatic factors and interindividual heterogeneity within each growth phase. On the basis of these integrative statistical models, it is shown that walnut saplings have an opportunistic mode of development that is primarily driven by the changing light environment. In particular, light availability explains the ability of a tree to reach a phase of strong growth where the first branches can appear. It is also shown that growth fluctuation amplitudes in response to climatic factors increased while interindividual heterogeneity decreased during tree development.     

LACK OF DEFENSE OR PHLOROTANNIN INDUCTION BY UV RADIATION OR MESOGRAZERS IN DESMARESTIA ANCEPS AND D. MENZIESII (PHAEOPHYCEAE)1

Phlorotannins are polyphenoloic metabolites occurring only in the Phaeophyceae that have numerous putative primary roles (e.g. cell‐wall construction and storage) as well as secondary metabolic roles, which include herbivore feeding deterrence and protection from UV radiation. The proposed role of phlorotannins in the defense against UV radiation is of particular importance in the Antarctic due to depletion of the stratospheric ozone layer in that area. Several studies of brown algae have found evidence of an induction response (the production of defensive metabolites, including phlorotannins) after grazing by various mesograzers, after simulated grazing/wounding, and after exposure to increases in UV radiation. This study aimed to determine if phlorotannin production or other defenses in two dominant, endemic Antarctic species (Desmarestia menziesii Montagne and Desmarestia anceps J. Agardh) could be induced by an increase in exposure to UV radiation or by natural and artificial grazing. An in situ experiment failed to detect any effect of UV radiation on phlorotannin concentrations in either species or on subsequent palatability in feeding bioassays. A laboratory‐based experiment did not detect any effect of mesoherbivore grazing or simulated grazing (wounding) on palatability or the concentration of phlorotannins in D. menziesii. Instead, phlorotannin concentrations increased in all treatments in both experiments, consistent with an increase in overall resource availability due to an increase in available PAR compared with the in situ irradiance at the algal collection sites.     

AXENIC TISSUE CULTURE AND MORPHOGENESIS IN PORPHYRA YEZOENSIS (BANGIALES, RHODOPHYTA)

To better understand developmental phenomena in macroalgal tissue culture, we examined the morphogenesis of Porphyra yezoensis Ueda (strain TU-1) cultured aseptically in defined synthetic media . Generally, the filamentous thalli (sporophyte; conchocelis phase) of P. yezoensis were densely tufted with uniseriate filaments. The foliose thalli (gametophyte) were monolayered. In this study, axenic filamentous thalli retained their characteristic morphogenesis; there were no obvious differences between morphogenetic traits in unialgal and axenic conditions. However, conchospores, which might have developed into the foliose form under unialgal conditions, germinated into calluslike masses under axenic conditions. Most of the gametophytes gradually lost their typical morphogenesis after the first longitudinal cell division. Some of the calluslike masses developed rhizoidlike structures in several places or along the entire mass. Therefore, we concluded that P. yezoensis, in axenic cultures, loses its typical morphogenesis only during the gametophytic phase. The axenic tissue culture of Porphyra established in this study is a promising assay system for the identification of growth and morphogenetic factors.     

Survival and growth of alpha-spores of Porphyra schizophylla Hollenberg (Rhodophyta: Bangiophyceae)

Alpha-spores of the intertidal Porphyra schizophylla Hollenberg f. norrisii Mumford appear able to tolerate environmental stress factors associated with intertidal habitats, (viz., drying of water films with salt crystal formation, increased salt concentrations in the surrounding water, exposure to humid air when spores are enclosed in water films), and to produce Conchocelis filaments of normal appearance when subsequently germinated in sea water cultures. Tolerances of lowered salinities were observed down to 17.4%., but little survival at salinities lower than this. Alpha-spore germination on thalli of intertidal macro-algae was observed, with initial penetration in some cases. Penetrations of algal thalli were observed with Conchocelis filaments of α-spores germinated on red algae from low intertidal and subtidal habitats. Cell recognition phenomena would seem to be involved in these growth responses.     

Growth kinetics related to physiological parameters in young Saccorhiza dermatodea and Alaria esculenta sporophytes exposed to UV radiation

Young sporophytes of Saccorhiza dermatodea and Alaria esculenta cultured from Spitsbergen isolates were exposed in the laboratory to either only photosynthetically active radiation (PAR) or to a spectrum including UV-radiation (PAR+UVA+UVB) by use of cutoff glass filters. The plants were grown at 8±2°C and 16:8 h light–dark cycles with 6 h additional UV exposure in the middle of the light period. Growth was measured every 10 min using growth chambers with online video measuring technique for 18–21 days. Tissue morphology and absorption spectra were measured in untreated young sporophytes while tissue chlorophyll-a content and DNA damage were measured from treated thalli at the end of the experiment. Under UVR, growth rates of S. dermatodea were significantly reduced while A. esculenta have a potential to acclimate. Tissue chlorophyll-a contents in both species were not significantly different between treatments suggesting that these algae may acclimate to moderate UVR fluence. Higher DNA damage in S. dermatodea effectively diverted photosynthetic products for repair constraining growth. Tissue optics (opacity and translucence) was correlated to the tissue absorbance in the UVR region characteristics of phlorotannin, an important UV-absorbing compound in brown macroalgae. Growth rates of sporophytes of both species exposed to PAR without UV was similar during day and night. The results showed that both species can recruit and inhabit a similar coastal zone when appropriate strategies are expressed to minimize damage in response to the stress factor.