UV-absorbing substances in zooxanthellate and azooxanthellate clams

The effects of UV-A and UV-B radiation on photosynthesis of zooxanthellae within the siphonal mantle of the giant clam, Tridacna crocea, and in isolation were studied. While UV-B irradiation (2.4 W m⁻², 20 min) completely suppressed photosynthesis of the isolated zooxanthellae, it had little effect on their photosynthetic ability if they were irradiated while within the siphonal mantle of the host tissue. Chemical analysis of the siphonal mantle of T. crocea showed the presence of significant amounts of mycosporine-like amino acids (MAAs), which absorb UV-A and -B light. However, no MAA was detected in the isolated zooxanthellae. MAAs were concentrated in the siphonal mantle and kidney tissues in comparison with other tissues. In the siphonal mantle, MAA concentrations were the highest in the outermost surface layer where most of the zooxanthella cells resided. This indicates that the zooxanthellae are protected from UV radiation by a screen of concentrated MAAs in the host clam. Aside from T. crocea, significant amounts of MAAs were found not only in other zooxanthellate clams, such as T. derasa, Hippopus hippopus, Colculum cardissa and Fragum unedo, but also in a closely related azooxanthellate clam, Vasticardium subrugosum. On the other hand, no MAA was detected in any of the zooxanthellae from these zooxanthellate clams. No MAA was detected in the tissues of a deep-sea bivalve, Calyptogena soyoae. Although MAAs seem to block strong UV radiation in the shallow-water clam, they are probably not essential for the clam's life in the dark. MAAs in shallow-water clams may be derived from food and accumulated in their tissues, especially in the siphonal mantle and kidney. Received: 29 November 1996 / Accepted: 13 January 1997     

Development and survivorship of zooxanthellate and azooxanthellate primary polyps of the soft coral Heteroxenia fuscescens: laboratory and field comparisons

Marine Biology - The zooxanthellate Red Sea soft coral Heteroxenia fuscescens releases aposymbiotic planulae nearly all year round, with higher numbers in the summer than in other seasons. After...     

丛枝菌根真菌砷酸盐还原酶基因RiarsC的克隆和功能分析

丛枝菌根真菌(AMF)在增强植物砷(As)抗性方面发挥着重要作用。已有相关研究表明,接种AMF能提高植物体内三价砷As(III)的比例,AMF可能参与了将五价砷As(V)还原为As(III)的过程从而提高了菌根植物的As抗性,但目前尚缺乏直接分子证据。本文从异形根孢囊霉(Rhizophagus irregularis)菌丝中克隆得到了一个砷酸盐还原酶基因RiarsC并进行序列分析。将该基因转入arsC缺陷型大肠杆菌(Escherichia coli)菌株WC3110(ΔarsC)中,通过As(V)抗性生长曲线和As形态测定,分析了该基因的功能。结果显示,RiarsC属于谷氧还蛋白-谷胱甘肽依赖的砷酸盐还原酶家族;RiarsC基因的表达显著提高了As敏感型E.coli菌株对As(V)的抗性,当培养基中As(V)浓度为100μmol·L-1时表现更加明显。As形态分析表明,表达RiarsC的E.coli菌株能够将培养基中71.03%的As(V)还原为As(III);与表达空载体的菌株相比,还原效率提高了61.98%。本研究证明了AMF的砷解毒还原能力,为进一步开展AMF的砷代谢机制研究提供了一定的分子生物学基础。     

Size and spatial structure in deep versus shallow populations of the Mediterranean gorgonian <Emphasis Type="Italic">Eunicella singularis</Emphasis> (Cap de Creus,northwestern Mediterranean Sea)

In the Western Mediterranean Sea, the gorgonian Eunicella singularis (Esper, 1794) is found at high densities on sublittoral bottoms at depths from 10 to 70 m. Shallow colonies have symbiotic zooxanthellae that deeper colonies lack. While knowledge of the ecology of the shallow populations has increased during the last decades, there is almost no information on the ecology of the deep sublittoral populations. In October and November 2004 at Cap de Creus (42°19′12″ N; 03°19′34″ E), an analysis of video transects made by a remotely operated vehicle showed that shallow populations (10–25 m depth) were dominated by small, non-reproductive colonies, while deep sublittoral populations (50–67 m depth) were dominated by medium-sized colonies. Average and maximum colony heights were greater in the deeper populations, with these deeper populations also forming larger patch sizes and more extensive regions of continuous substrate coverage. These results suggest that shallow habitats are suitable for E. singularis, as shown by the high recruitment rate, but perturbations may limit or delay the development of these populations into a mature stage. This contrasts with the deep sublittoral habitats where higher environmental stability may allow the development of mature populations dominated by larger, sexually mature colonies.     

Comparative study of the chemistry and structure of gorgonian and antipatharian coral skeletons

The comparative morphology of the axial skeleton was studied in representatives of 5 reef-dwelling gorgonian families and 2 antipatharian genera by electron and light microscopy. Comparative axial histochemistry, amino acid composition and the physico-chemical character of the protein were also studied. Evidence obtained indicates that collagen is a widespread and prominent structural feature of the gorgonian axial skeleton. Aromatic crosslinkage (sclerotization) also appears to be of widespread, if not universal occurrence in these animals. Sclerotization is equally characteristic of the black coral skeletons studied, but this material does not contain collagen. Its unusual composition, structure, and reactivity is discussed.     

Characterization and role of carbonic anhydrase in the calcification process of the azooxanthellate coral <Emphasis Type="Italic">Tubastrea aurea</Emphasis>

In zooxanthellate corals, the photosynthetic fixation of carbon dioxide and the precipitation of CaCO₃ are intimately linked both spatially and temporally making it difficult to study carbon transport mechanisms involved in each pathway. When studying Tubastrea aurea, a coral devoid of zooxanthellae, we can focus on carbon transport mechanisms involved only in the calcification process. We performed this study to characterize T. aurea carbonic anhydrase and to determine its role in the calcification process. We have shown that inhibition of tissular carbonic anhydrase activity affects the calcification rate. We have measured the activity of this enzyme both in the tissues and in the organix matrix extracted from the skeleton. Our results indicate that organic matrix proteins, which are synthesized by the calcifying tissues, are not only structural proteins, but they also play a crucial catalytic role by eliminating the kinetic barrier to interconversion of inorganic carbon at the calcification site. By immunochemistry we have demonstrated the presence of a protein both in the tissues and in the organic matrix, which shares common features with prokaryotic carbonic anhydrases.     

食蚊鱼(Gambusia afinis)cat、gapdh和gst基因的克隆及其在生态毒理学中的应用

根据Ensembl、Genbank登录的鱼类cat、gapdh和gst基因的CDS序列设计普通PCR扩增引物,寻找食蚊鱼的cat、gapdh和gst基因的c DNA片段,并根据定量引物设计要求设计出相应的SYBR Green I荧光定量RT-q PCR引物,建立了食蚊鱼cat、gapdh和gst基因的SYBR Green I荧光定量RT-q PCR方法。该方法在104~108数量级范围内有良好线性关系(R=0.999~1.000);熔解曲线显示扩增产物特异性良好,均为单一峰值;质粒标准品最高浓度与最低浓度的批内试验变异系数与批间试验变异系数均低于2%。利用该方法监测和评价环境污染物对水生生物的影响,选择了水体中常见典型药物污染物——双氯芬酸,研究其对食蚊鱼抗氧化基因表达的影响。结果表明,雌性食蚊鱼暴露在不同浓度双氯芬酸钠(0.005、0.05、0.5和5 mg·L-1)24 h后,其肝脏cat、gapdh和gst的mRNA呈现显著变化,相对于对照组,在低浓度0.005 mg·L-1时,cat与gst mRNA的表达量均有极显著上升(p0.01),而其它浓度均极显著下降(p0.01)。试验表明该方法具有快速、精确、灵敏度高的优点,可为利用该类小型鱼类的原位污染物的生物监测和生态毒理评价提供有力的技术支持。     

淡水双壳类背角无齿蚌AwHSP70基因克隆及多溴联苯醚-47对其表达的影响

热休克蛋白(HSP)参与蛋白质折叠、细胞膜转位和错误折叠蛋白质降解等过程,提高动物对环境的应激能力和适应能力。前期研究表明,多溴联苯醚-47(PBDE-47)对背角无齿蚌具有显著的氧化应激和急性毒性效应,为探讨PBDE-47慢性毒性效应;将背角无齿蚌随机分为对照组和PBDE-47处理组,处理组用3.36μg·L~(-1)浓度的PBDE-47进行处理,对照组用相同体积的二甲亚砜进行处理;克隆出AwHSP70基因,分析PBDE-47对AwHSP70表达的影响。结果显示,AwHSP70具有HSP70家族的标签序列,广泛分布于斧足、鳃、肝胰脏、闭壳肌、心脏、血淋巴和外套膜。PBDE-47处理可导致肝胰脏、鳃和血细胞中AwHSP70 mRNA水平显著升高。与对照组相比,PBDE-47处理组肝胰脏中AwHSP70 mRNA水平在1～15 d内增加2. 79倍(P<0.01)以上;鳃中AwHSP70 mRNA水平增加3.06倍(P<0.01)以上;血淋巴中AwHSP70表达增加1.81倍(P<0.05)以上。背角无齿蚌上调AwHSP70表达有助于增强动物对PBDE-47的耐受能力。     

Depth-dependent responses to solar ultraviolet radiation and oxidative stress in the zooxanthellate coral Acropora microphthalma

Colonies of Acropora microphthalma (Verrill 1869) were transferred from depths of 2 to 3, 10, 20, and 30 m to UV-transparent and UV-opaque respirometry chambers placed at 1 m depth at Bowl Reef, Great Barrier Reef, in March 1989. Peak rates of photosynthesis in colonies originating at 2 and 10 m were unaffected by solar ultraviolet (UV) radiation at 1 m, whereas photosynthesis showed 30 and 38% inhibition in colonies transferred from 20 and 30 m, respectively. This differential sensitivity of corals to UV radiation was consistent with the five- to tenfold higher concentrations of UV-absorbing, mycosporine-like amino acids (MAAs, putative defenses against UV) in 2- and 10-m colonies compared with 20- and 30-m colonies. Photosynthesis in zooxanthellae freshly isolated from 2- and 10-m corals, however, was inhibited by UV, indicating that the host's tissues, which contain 95% of the total MAAs in corals at these depths, are the first line of defense against solar UV and provide protection to their endosymbiotic algae. The general bathymetric decline in the activities of the antioxidant enzyme superoxide dismutase (SOD) in the host, and SOD, catalase, and ascorbate peroxidase in the zooxanthellae, is related to the decrease in potential for photooxidative stress with increasing depth.     

Photoinhibition of Symbiodinium spp. within the reef corals Montastraea faveolata and Porites astreoides: implications for coral bleaching

It is speculated that differences in coral bleaching susceptibility may be influenced by the genotype of in hospite Symbiodinium and their differential responses to bleaching stressors. Photoinhibition of photosystem II (PSII), damage to the D1 (psbA) PSII reaction centre protein and production of reactive oxygen species by in hospite Symbiodinium are likely precursors of coral bleaching. In order to assess whether photorepair rates of in hospite Symbiodinium underlie the bleaching susceptibility of their hosts, photoinhibition (net and gross), photoprotection and photorepair rates were assessed in a bleaching-‘tolerant’ coral (P. astreoides) and a bleaching-‘sensitive’ coral (M. faveolata) using non-invasive fluorometric techniques and by blocking de novo synthesis of psbA. Previous studies using such techniques have demonstrated that in vitro Symbiodinium types ‘sensitive’ to bleaching stressors had reduced rates of photorepair relative to ‘tolerant’ Symbiodinum types. Our measurements demonstrated that Symbiodinium in the more bleaching tolerant P. astreoides had higher photorepair rates than Symbiodinium in M. faveolata. Higher repair rates in P. astreoides resulted in lower net photoinhibition relative to M. faveolata, where both corals exhibited similar susceptibility to photodamage (gross photoinhibition). Photoprotective mechanisms were observed in both corals; M. faveolata exhibited higher antennae-bed quenching than P. astreoides at low-light intensities, but at and above light-saturating intensities, which are different for each coral species, P. astreoides displayed more efficient non-photochemical quenching (Stern–Volmer quenching) of chlorophyll fluorescence than M. faveolata. Increased NPQ by P. astreoides at E/E _k ≥ 1 was not driven by antennae-bed quenching. The ability of in hospite Symbiodinium in P. astreoides to mitigate the effects of photoinhibition under high light conditions compared with Symbiodinium in M. faveolata, and their high repair capacity following photoinhibition, may be a key factor to consider in future bleaching studies and may underlie the relative bleaching tolerance of P. astreoides compared to M. faveolata.     

Photosynthesis, respiration and calcification of a zooxanthellate scleractinian coral under submerged and exposed conditions

An experimental set-up was designed to investigate photosynthesis, respiration and calcification of zooxanthellate scleractinian corals under submerged and exposed conditions. The results of experiments to determine the effect of exposure to air on the metabolism of Stylophora pistillata (Esper, 1797) revealed that: (1) maximum gross photosynthesis ( p _{g max}) is 3.6 times higher in water than in air; (2) an indicator of photoadaptation (I _k ) does not reveal any difference between water and air; (3) the difference between submerged and aerial respiration is not statistically significant; (4) aerial photosynthesis–irradiance (P–I) curves display lower initial slopes (α) than aquatic P–I curves; (5) there is no calcification in air. Received: 7 February 1997 / Accepted: 4 March 1997     

Temporal changes in the trophic ecology of the asymbiotic gorgonian Leptogorgia virgulata

Gorgonians are widespread sub-littoral benthic suspension feeders in the world oceans. However, data on their trophic ecology and role in benthic–pelagic coupling and biogeochemical cycles remain limited. This study assesses the trophic ecology of Leptogorgia virgulata in the Skidaway River estuary, USA (31.9896N, 81.0242W) by analysing carbon and nitrogen isotopic signatures of its soft tissue and different-sized fractions of particulate organic matter (POM) in its environment. Samples were taken in 5 April, 2 August, and 15 October 2012 and 11 January 2013. Results support a distinct temporal shift in the diet of L. virgulata from the POM fraction <10 µm (i.e. pico- and nanoplankton) to the 10–63-µm fraction (i.e. microplankton). This trophic regime is likely associated with the natural abundance of prey items within these size classes, thus suggesting that L. virgulata may be an opportunistic feeder, and seasonal shifts in food availability in the water column affect its diet. As such small prey items affect the bioenergetics of L. virgulata, it is important to understand the implications of changes in food availability associated with environmental drivers on the physiology and population dynamics of this dominant species in the western Atlantic Ocean.     

Possible control mechanism of cell motility in the gorgonian Eunicella cavolinii

In a previous study it was demonstrated that a lectin controls cell-cell interaction in the gorgonian Eunicella cavolinii (Koch) as a negative modulator. Now we describe the procedure to purify this lectin to homogeneity; its molecular weight is 23 400. The homologous proteoglycans were identified as positive modulators of cell-cell (and/or cell substrate) interaction. The purified single proteoglycan aggregates were 1200±700 nm long and the distance between the attachment points of the proteoglycan subunits was about 45 nm. The glycosaminoglycan residues of the gorgonian proteoglycans were identified as hyaluronic acid (35.5%), heparan sulfate (47.9%) and dermatan sulfate (14.1%). Binding studies with immobilized homologous proteoglycan preparations revealed that gorgonian cells attach to this substratum presumably via its glycosaminoglycans. Quantitative determinations of these two modulators of cell-cell recognition in the three regions of individual colonies revealed concentration gradients these concentration differences reflect a lower cell motility in the polyp-bearing branches and a higher motility of the cells in the polyp-free stem and in the basis, is discussed.     

Chemical changes accompanying maturation of the connective tissue skeletons of gorgonian and antipatharian corals

The skeletons of 3 gorgonian and 2 antipatharian corals were subjected to quantitative chemical analysis. Protein values ranged from 70.4 to 93.6%; ash from 3.0 to 19.4%; lipid from 0 to 8.4%; carbohydrate from 1.24 to 3.94% and halogen from 4.2 to 24.9% of the dry skeletal weight. Hydroxyproline and phenolic values were 0 to 3.0% and 11.6 to 25.0% of the protein, respectively. Lipid, present in 2 gorgonian species and 1 antipatharian, significantly decreased with age in all three cases. Glucose and galactose accounted for over 90%, and sialic acids for an additional 1 to 10%, of the carbohydrate in the gorgonian skeletons studied; the glucose content of the gorgonian skeleton decreased with age. The antipatharian skeletons possessed no glucose or galactose, but contained significant levels of amino sugars; the presence of chitin is confirmed. In the gorgonians, bromine and iodine, the predominant halogens, increased with skeletal age and were present in nearly equal amounts. Small amounts of bromine were found in the antipatharian skeletons, but very large amounts of iodine were found in older parts of the skeleton. The basal regions of both antipatharian species contained >23% by weight of iodine, the highest content of iodine yet recorded for any organism.     

Role of carbonic anhydrase in the supply of inorganic carbon to the giant clam—zooxanthellate symbiosis

The mechanism whereby inorganic carbon (C_i) is acquired by the symbiotic association between the giant clam (Tridacna derasa) and zooxanthellae (Symbiodinium sp.) has been investigated. C_i in the haemolymph of the clam is in equilibrium with the surrounding sea water. The photosynthesis rate exhibited by the intact clam varies as a function of the C_i concentration in the clam haemolymph. The gill tissue contains high carbonic anhydrase activity which may be important in adjusting the C_i equilibrium between haemolymph and sea water. Zooxanthellae (Symbiodinium sp.) isolated from the clam mantle prefer CO₂ to HCO ₃ ^- as a source of inorganic carbon. The zooxanthellae have low levels of carbonic anhydrase on the external surface of the cell; however, mantle extracts display high carbonic anhydrase activity. Carbonic anhydrase is absent from the mantle of aposymbiotic clams (T. gigas), indicating that this enzyme may be essential to the symbiosis. The enzyme is probably associated with the zooxanthellae tubes in the mantle. The results indicate that carbonic anhydrase plays an important role in the supply of carbon dioxide within the clam symbiosis.