Analysis of DNA Methylation of <Emphasis Type="Italic">Gracilariopsis lemaneiformis</Emphasis> Under Temperature Stress Using the Methylation Sensitive Amplification Polymorphism (MSAP) Technique

Gracilariopsis lemaneiformis is an economically important agarophyte, which contains high quality gel and shows a high growth rate. Wild population of G. lemaneiformis displayed resident divergence, though with a low genetic diversity as was revealed by amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) analyses. In addition, different strains of G. lemaneiformis are diverse in morphology. The highly inconsistence between genetic background and physiological characteristics recommends strongly to the regulation at epigenetic level. In this study, the DNA methylation change in G. lemaneiformis among different generation branches and under different temperature stresses was assessed using methylation sensitive amplified polymorphism (MSAP) technique. It was shown that DNA methylation level among different generation branches was diverse. The full and total methylated DNA level was the lowest in the second generation branch and the highest in the third generation. The total methylation level was 61.11%, 60.88% and 64.12% at 15°C, 22°C and 26°C, respectively. Compared with the control group (22°C), the fully methylated and totally methylated ratios were increased in both experiment groups (15°C and 26°C). All of the cytosine methylation/demethylation transform (CMDT) was further analyzed. High temperature treatment could induce more CMDT than low temperature treatment did.     

Inhibition of photosynthesis in the microalga Chaetoceros curvisetus （Bacillariophyta） by macroalga Gracilaria lemaneiformis （Rhodophyta）

We investigated the effects of dried macroalga Gracilaria lemaneiform （Rhodophyta） on photosynthesis of the bloom-forming microalga Chaetoceros curvisetus. C. curvisetus was cultured with different amounts of dried G. lemaneiformis under controlled laboratory conditions. We measured the photosynthetic oxygen evolution rate and established the chlorophyll a fluorescence transient （OJIP） curve coupled with its specific parameters. We observed concentration-dependent and time-dependent relationships between dried G. lemaneiformis and inhibition of photosynthesis in C. curvisetus. Co-culture with dried G. lemaneiformis also resulted in a decrease in the light-saturated maximum photosynthetic oxygen evolution rate （P~ax） in C. curvisetus, and a decrease in the OJIP curve along with its specific parameters; the maximum photochemical efficiency of PSII （FJFm）, the amount of active PSII reaction centers per excited cross section at t=0 and t=--tFM （RC/CS0 and RC/CSm, respectively）, the absorption flux per excited cross section at t=0 （ABS/ CS0）, and the efficiency with which a trapped exciton moves an electron into the electron transport chain （~u0）. The dark respiration rate （Rd） increased in C. curvisetus co-cultured with dried G. lemaneiformis. The JIP-test and the oxygen evolution results indicated that dried G. lemaneiJbrmis decreased the number of active reaction centers, blocked the electron transport chain, and damaged the oxygen-evolving complex of C. curvisetus. This result indicated that dried fragments of G. lemaneiformis could effectively inhibit photosynthesis of C. curvisetus, and thus, could serve as a functional product to control and mitigate C. curvisetus blooms.     

Bioreactor technology for production of valuable algal products

INTRODUCTIONManystudiesonvascularplantSduringthenontwperiodhavedrireP0redinChinaandabroad.TherearefeWreportSonwhethertherearesomevasctilarplantS,andhowmanyspedessurvive,underice,andontheirbiomassesandphotosyntheticproduCtivitiesduringwinter,espedal1yduringbeAfpinchLillyamepe0ylenctal.,l976,Liuetal.,l994).WeimtlycarriedoutaseadriontheunderforvascularplantsdurinwinderinsomewaterbodieswherevascularplantSgrowluxdrianhydurinInildareonsinnorthemareas,soastoimproveourknowedgeofvascularplaflt…     

Effects of temperature, algae biomass and ambient nutrient on the absorption of dissolved nitrogen and phosphate by Rhodophyte Gracilaria asiatica

Gracilaria asiatica,being highly efficient in nutrient absorption,is cultivated in sea cucumber ponds to remove nutrients such as nitrogen and phosphate.It was cultured in a laboratory simulating field conditions,and its nutrient absorption was measured to evaluate effects of environmental conditions.Ammonia nitrogen(AN),nitrate nitrogen(NN),total inorganic nitrogen(TIN),and soluble reactive phosphorus(SRP) uptake rate and removal efficiency were determined in a 4×2 factorial design experiment in water temperatures(T) at 15℃ and 25℃,algae biomass(AB) at 0.5 g/L and 1.0 g/L,total inorganic nitrogen(TIN) at 30 μmol/L and 60 μmol/L,and soluble reactive phosphorus(SRP) at 3 and 6 μmol/L.AB and ambient TIN or SRP levels significantly affected uptake rate and removal efficiency of AN,NN,TIN,and SRP(P<0.001).G.asiatica in AB of 0.5 g/L showed higher uptake rate and lower removal efficiency relative to that with AB of 1.0 g/L.Nitrogen and phosphorus uptake rate rose with increasing ambient nutrient concentrations;nutrient removal efficiency decreased at higher environmental nutrient concentrations.The algae preferred to absorb AN to NN.Uptake rates of AN,NN,and SRP were significantly affected by temperature(P<0.001);uptake rate was higher for the 25℃ group than for the 15℃ group at the initial experiment stage.Only the removal efficiency of AN and SRP showed a significant difference between the two temperature groups(P<0.01).The four factors had significant interactive effects on absorption of N and P,implying that G.asiatica has great bioremedial potential in sea cucumber culture ponds.     

Biomass and carbon storage of Gracilariopsis lemaneiformis （Rhodophyta） in Zhanshan Bay,Qingdao, China

Marine macroalgae can absorb carbon and play an important role in carbon sequestration. As an important economic macroalga, Gracilariopsis lemaneiformis has the potential to significantly affect carbon absorption and storage in wave-sheltered intertidal reef systems. However, detailed knowledge on seasonal biomass changes and carbon storage of G. lemaneiformis is lacking, especially in many small and scattered ecosystems. Considering the influence of human activities on wild distribution of G. lemaneiformis, the understanding of seasonal dynamics of an economically important species in nature is necessary. In this study, we first investigated seasonal variations in biomass, coverage area, and carbon storage during low tide from August 2011 to July 2012 in Zhanshan Bay, Qingdao, China. Furthermore, we estimated the carbon storage potential of wild G. lemaneiformis using light use efficiency(LUE). The results show that the standing biomass and coverage area changed significantly with season. However, seasonal variations in carbon content and water content were not obvious, with an average content of 35.1% and 83.64%, respectively. Moreover, carbon storage in individual months varied between 0.67 and 47.03 g C/m 2, and the value of carbon storage was the highest in August and June and the lowest in February. In Zhanshan Bay, LUE of G. lemaneiformis was only 0.23%. If it is increased to the theoretical maximum(5%–6%), the carbon storage will have an increase of at least 21 times compared with the current, which suggested that carbon storage of wild G. lemaneiformis had a high enhancement potential. The study will help to assess a potential role of G. lemaneiformis in reducing atmospheric CO2.     

Genome size of <Emphasis Type="Italic">Alexandrium catenella</Emphasis> and <Emphasis Type="Italic">Gracilariopsis lemaneiformis</Emphasis> estimated by flow cytometry

Flow cytometry(FCM) technique has been widely applied to estimating the genome size of various higher plants. However, there is few report about its application in algae. In this study, an optimized procedure of FCM was exploited to estimate the genome size of two eukaryotic algae. For analyzing Alexandrium catenella, an important red tide species, the whole cell instead of isolated nucleus was studied, and chicken erythrocytes were used as an internal reference. The genome size of A. catenella was estimated to be 56.48 ± 4.14 Gb(1C), approximately nineteen times larger than that of human genome. For analyzing Gracilariopsis lemaneiformis, an important economical red alga, the purified nucleus was employed, and Arabidopsis thaliana and Chondrus crispus were used as internal references, respectively. The genome size of Gp. lemaneiformis was 97.35 ± 2.58 Mb(1C) and 112.73 ± 14.00 Mb(1C), respectively, depending on the different internal references. The results of this research will promote the related studies on the genomics and evolution of these two species.     

Comparison in copper accumulation and physiological responses of Gracilaria lemaneiformis and G. lichenoides (Rhodophyceae)

Heavy metal pollution has become a worldwide problem in aquaculture. We studied copper (Cu2+ ) accumulation and physiological responses of two red algae Gracilaria lemaneiformis and Gracilaria lichenoides from China under Cu2+ exposure of 0-500 μg/L in concentration. Compared with G. lemaneiformis, G. lichenoides was more capable in accumulating Cu2+ , specifically, more Cu2+ on extracellular side (cell wall) than on intracellular side (cytoplasm) and in cell organelles (especially chloroplast, cell nucleus, mitochondria, and ribosome). In addition, G. lichenoides contained more insoluble polysaccharide in cell wall, which might promote the extracellular Cu2+ -binding as an efficient barrier against metal toxicity. Conversely, G. lemaneiformis was more vulnerable than G. lichenoides to Cu 2+ toxin for decreases in growth, pigment (chlorophyll a, chlorophyll b, phycobiliprotein, and β-carotene) content, and photosynthetic activity. Moreover, more serious oxidative damages in G. lemaneiformis than in G. lichenoides, in accumulation of reactive oxidative species and malondialdehyde, and in electrolyte leakage, because of lower antioxidant enzyme (superoxide dismutase and glutathione reductase) activities. Therefore, G. lichenoides was less susceptible to Cu2+ stress than G. lemaneiformis.     

A comparison of different Gracilariopsis lemaneiformis (Rhodophyta) parts in biochemical characteristics,protoplast formation and regeneration

Gracilariopsis lemaneiformis is a commercially exploited alga. Its filaceous thallus can be divided into three parts, holdfast, middle segment and tip. The growth and branch forming trend and agar content of these three parts were analyzed, respectively, in this study. The results showed that the tip had the highest growth rate and branched most, although it was the last part with branch forming ability. The holdfast formed branches earliest but slowly. Holdfast had the highest agar content. We also assessed the difference in protoplast formation and regeneration among three parts. The middle segment displayed the shortest enzymolysis time and the highest protoplast yield; whereas the tip had the strongest vitality of protoplasts formation. Juvenile plants were only obtained from the protoplasts generated from the tip. These results suggested that the differentiation and function of G. lemaneiformis was different.     

Main nutritional contents of 30 Dalian coastal microalgae species

This paper reports results of study on the contents of proteins, amino acids, polysaccharose and uronic acids in 30 species of macroalgae from Shicao, Heishijiao, Shimiao, and Xiaofujiazhuang in the vicinity of Dalian City, N.E.China. The results showed that the protein contents of the 30 algae from highest (112.55μg/ml) to the lowest (0.24 p.g/ml) was in the descending order of Dictyopteris ndalata, Gelidium vagum,Gymnogongrus japonican, Ectocarpus confervoides, Tinocladia crassa, Sargassum thunberii. In general, the protein content in red algae was higher than that in brown algae. The content of free ami no acids showed nosignificent differences from 7.44μg/ml-4.96μg/ml in all these algae, in the descending order of Gymnogongrusrus japonican, Sargassum confusum, Undoria pinnatifida, Laminaria japonica and Ectocarpus confervoides. The content of polysaccharose varied from 168.2 gg/ml-22.15 p.g/ml in the descending order of Symphyocladiaia tiuscula, Scytosiphon lomentarius, Desmarestia viridis., Tinocladia crassa, Gracilaria asiatica and Porphyra yezoensis. The content of uronic acids is from 196.24μg/ml-20.77μg/ml in the descending order of Ulva lactuca,Symphyoclaldia latiuscula, Scytosiphon lomentarius, Ceramimum kondoi, Gracilaria vemucosa and Porphyra yezoensis. The fatty acids in 30 species of algae belong to Rhodophyta, Chlorophyta and Phaeophyta. Most phaeophytes have many (4-12) types of fatty acids.     

Phylogenetic analysis of epibacterial communities on the surfaces of four red macroalgae

Macroalgal surfaces are prone to being attached by bacteria. Epibacterial community structures on marine macroalgae are host-specific but temporally and spatially variable. In this study, we investigated the structure of epibacterial communities on the surfaces of four red macroalgae, Gracilaria lemaneiformis, Gloiopeltis furcata, Mazzaella sp. and Porphyra yezoensis, by analyzing the sequences of 16 S r RNA gene libraries. Healthy individuals of all macroalgae species were collected in winter from a farm at Dalian, China. The results showed that the epibacterial communities were mainly dominated by α-Proteobacteria, γ-Proteobacteria and Bacteroidetes. Deinococcus-Thermus, Spirochaetes and ε-Proteobacteria were also found. The majority of cloned sequences shared the greatest similarity to those of culturable organisms. A large portion of sequences from the α-Proteobacteria homed in Roseobacter clade, i.e., genera Ahrensia, Roseovarius, Litoreibacter, Octadecabacter, Thaiassobacter and Sulfitobacter, while members of Bacteroidetes mainly belonged to family Flavobacteriaceae. The cloned sequences could be separated into 66 OTUs at 0.01 distance value, and rare common OTUs were found among libraries. At genus level, Pseudoalteromonas dominated Gr. lemaneiformis and Gl. furcata libraries, accounting for 72.2% and 47.3%, respectively. Sulfitobacter dominated P. yezoensis library, accounting for 35.4%. A previously undefined cluster within Deinococcus-Thermus dominated Mazzaella sp. library, accounting for 24.6% of the all. These results indicated that a broad range of bacteria inhabited the surfaces of these macroalgae.     

Study on the dominant species of Pteropoda in East China Sea

1 INTRODUCTION Pteropoda is a group of marine pelagic mol- lusks, which belongs to Opisthobranchia of Gastro- poda. Although Pteropoda is not a major group in pelagic zooplankton in terms of abundance and number of species, their ecological characters are…     

Simulation and analysis on seasonal variability of average salinityin the Yellow Sea

The CTD (conductivity, temperature and depth) data collected by six China-Korea joint cruises during 1996-1998 and the climatological data suggest that the seasonal variability of average salinity in the Yellow Sea (Sa) presents a general sinusoid pattern. To study the mechanism of the variability, annual cycles of Sa were simulated and a theoretical analysis based on the governing equations was reported.Three main factors are responsible for the variability: the Yellow Sea Warm Current (YSWC), the Changji-ang (Yangtze) River diluted water (YRDW) and the evaporation minus precipitation (E-P). From December to the next May, the variability of Sa is mainly controlled by the salt transportation of the YSWC. But in early July, the YSWC is overtaken and replaced by the YRDW which then becomes the most important controller in summer. From late September to November, the E-P gradually took the lead. The mass exchange north of the 37癗 line is not significant.     

Episodes of volcanic activity and their environmental effects in the Okinawa Trough during the last 150 ka

1 INTRODUCTIONA large-scale volcanic eruption can injectenormous amount of ash and dust into the atmos-phere, leading to local and short cooling due toshielding incoming radiation. During the past 2 to 3decades, accumulated evidence indicated thatlarge-scale volcano eruptions or series of volcanicactivity appears to coincide with climatologicaltransitions. For example, Robock (1979) indicatedthat volcanic eruptions played a role in forcing "Lit-tle Ice Age". Well-dated high-resolution co…     

Use of rapd for detecting and identifyingPorphyra (Bangiales, Rhodophyta)

Randomly amplified polymorphic DNA (RAPD) technique was applied to assess the genetic variations and phylogenetic relationships in 4 species ofPorphyra. The samples were collected from the coast of Canada, Vietnam, Zhoushan, Fujian and Qingdao in China. Amplifications with 20 primers were carried out under predetermined optimal reaction conditions (samples were first heated at 94°C for 5 min. and followed by 45 cycles of 1 min at 94°C, 1 min at 36°C, and 2 min at 72°C, then held at 72°C for 10 min). The amplified products were scored as present (1) or absent (0) for each DNA sample and an index of genetic similarity (F) was calculated by using Nei & Li’s matching coefficient method (1979). The value of (1−F) was used to quantify the genetic distances between species and construct a phylogenetic tree. The relationship indicated by the UPGMA and NJ cluster analysis on the values of the genetic distance is in good overall agreement with classical taxonomy. The obvious differences between natural and cultivated population ofP. haitanensi suggest that variation or hybridization with other species occurred during the culture. Contribution No. 100 from the Experimental Marine Biology Laboratory, IOCAS. Contribution No. 3289 from the Institute of Oceanology, Chinese Academy of Sciences. The research supported by the Foundation of the Chinese Academy of Sciences.     

Effect of temperature on gene expression in the pearl oyster Pinctada fucata

In this study, we examined the effect of elevated temperature on the expression patterns of genes, i.e., nacrein, irr, n16, n19, and hsp70 in the pearl oyster Pinctada fucata. The experiment was carried out at 4 temperatures, i.e., 20℃(ambient, control), 24, 28℃, and 32℃. The expression levels of target genes in P. fucata were assayed at 0, 6, 24, 48, and 96 h via real-time polymerase chain reaction. Results showed that the expression levels of nacrein and irr had no significant variations among different time points below 28℃, but significantly increased over time at 32℃. The expression levels of n16 and n19 did not change markedly at 20℃. The former increased significantly at 6 h and 24 h while the latter substantially decreased during 6–96 h at 24, 28 and 32℃. Among different temperatures, the level of n16 was significantly lower at 20℃ than at other temperatures during 6–96 h, and the level of n19 significantly varied among different temperatures at 48 h and 96 h. The expression level of hsp70 was significantly higher at 32℃ than at 20, 24 and 28℃ at 24 h. These results demonstrated that elevated temperature impacted the physiological processes of P. fucata and potentially influenced its adaptability to thermal stress.     

Distribution of dominant zooplankton species along a latitudinal gradient in China sea during spring

Dominant species of zooplankton community vary with latitude. Though China possesses a vast coastal area in northwestern Pacific, studies on the latitudinal dominant species gradient are rare. We collected zooplankton samples from Haizhou Bay(34.56?–35.19?N, 119.51?–120.30?E), Yueqing Bay(28.14?–28.38?N, 121.10?–121.21?E) and Dongshan Bay(23.65?–23.90?N, 117.45?–117.60?E) in May 2012 and May 2013 to preliminarily characterize the latitudinal dominant species distribution. All the samples were collected vertically using a 0.505 mm mesh plankton net with 0.8 m in mouth diameter from bottom to surface. Calanus sinicus, Aidanosagitta crassa, Labidocera euchaeta, Zonosagitta nagae, Acartia pacifica and Paracalanus parvus were found to be dominant. C. sinicus was the most dominant species and the unique one occurred in all three bays. With latitude decreasing, both the abundance and proportion of C. sinicus declined sharply. Cluster analysis showed that the 6 dominant species could be divided into 3 groups, based on their occurrences in the three bays. Our results suggested that the distribution of dominant species along the coast of China has a significant latitudinal gradient. C. sinicus which widely distributes in the coastal water of the northwestern Pacific can well adapt to the temperature at different latitudes. The high abundance in Haizhou Bay indicated that C. sinicus was an exemplary warm-temperate species, and more commonly occurs in the north of China seas. The ecological characteristics of dominant species change from warm-temperate type in high-latitudinal bays to warm water type in low-latitudinal bays.     

Simultaneous estimates ofsynechococcus spp. Growth and grazing mortality rates in the English Channel

The marine chroococooid phycoerythrin-containingSynechococcus spp. cyanobacterium has been implicated as a substantial component of the photosynthetic picoplankton in the ocean. Although its importance as food source for heterotrophic nanoplankton is now recognized, information about the cycling ofSynechococcus biomass and its diel pattern is limited and study methodology varies among authors. The selective metabolic inhibitor method was used to simultaneously estimate growth and grazing disappearance rates ofSynechococcus in the English Channel where growth rates ranged from 0.25 to 0.72/d (mean ±SD=0.51±0.17/d) and grazing mortality rates ranged from 0.19 to 0.64/d (mean ±SD=0.48±0.17/d). Size-fractionated experiments demonstrated that up to 70% ofSynechococcus disappearance could be attributed to grazers going through a 2 μm Nuclepore filter.Synechococcus grazing mortality rates (mean=0.74 ±0.25/d) during the day were always higher than that (mean=0.2±0.20/d) during the night, while growth rates showed no clear diel pattern. A positive correlation was observed between growth rates andin situ temperature ranging from 9 to 17°C, while in contrast grazing was independent of temperature. The close similatiry between average growth and grazing rates suggests a rapid recycling ofSynechococcus biomass in English Channel coastal waters.     

Current Vegetation Pattern along Glacial Landscape in Central （Garhwal） Himalaya, India

Introduction High mountain ecosystems are comparatively thrilling and sensitive at least at the upper elevation levels, and are determined by abiotic climate related ecological factors. Therefore, the ecosystems at the low temperature limits of plant life are generally considered to be particularly sensitive to climate changes (Koerner 1999). As temperature is a key factor for high mountain plants (Koerner and Larcher 1988, Gottfried et al. 1998), an upward migration of species must be conse…     

Growth,reproduction and population growth ofCeriodaphnia cornuta sars and comparison of 7-day fecundity withCeriodaphnia dubia richard

Biology of parthenogenetic females ofCeriodaphnia cornuta Sars was studied under laboratory conditions. The mean length of neonates (first-young instars) ofCeriodaphnia cornuta was found to be 0.27 mm at 25±1°C and that of the first adult instars 0.50 mm. A maximum length of 0.69 mm was attained at the 8th adult instar. The mean longevity was found to be 19.0 days for 3 preadult and 8 adult instars. Sex maturity was reached in 4.26 days. The mean number of young per brood was 6.21. The total production for each adult reached 49.7. The maximum reproduction period occurred during the 4th adult instar. At this age, the animals were 10.25 days old, and had a body length of 0.65 mm. The innate capacity for increase (r _m) was 0.40 per female per day, the net reproduction rate (R ₀) was 33.601 per generation, the mean length of a generation (T) was 8.786 days, and the finite rate for increase (λ) was 1.492 per female per day. Comparisons using the t-test indicated that the number of eggs produced and number of broods ofC. cornuta andC. dubia were not significantly different (p>0.05) after 7 days.