Production of polyhydroxybutyrate by the marine photosynthetic bacterium Rhodovulum sulfidophilum P5

The effects of different NaCl concentrations, nitrogen sources, carbon sources, and carbon to nitrogen molar ratios on biomass accumulation and polyhydroxybutyrate (PHB) production were studied in batch cultures of the marine photosynthetic bacterium Rhodovulum sulfidophilum P5 under aerobic-dark conditions. The results show that the accumulation of PHB in strain P5 is a growth-associated process. Strain P5 had maximum biomass and PHB accumulation at 2%-3% NaCl, suggesting that the bacterium can maintain growth and potentially produce PHB at natural seawater salinity. In the nitrogen source test, the maximum biomass accumulation (8.10±0.09 g/L) and PHB production (1.11±0.13 g/L and 14.62%±2.25% of the cell dry weight) were observed when peptone and ammonium chloride were used as the sole nitrogen source. NH 4 + -N was better for PHB production than other nitrogen sources. In the carbon source test, the maximum biomass concentration (7.65±0.05 g/L) was obtained with malic acid as the sole carbon source, whereas the maximum yield of PHB (5.03±0.18 g/L and 66.93%±1.69% of the cell dry weight) was obtained with sodium pyruvate as the sole carbon source. In the carbon to nitrogen ratios test, sodium pyruvate and ammonium chloride were selected as the carbon and nitrogen sources, respectively. The best carbon to nitrogen molar ratio for biomass accumulation (8.77±0.58 g/L) and PHB production (6.07±0.25 g/L and 69.25%±2.05% of the cell dry weight) was 25. The results provide valuable data on the production of PHB by R. sulfidophilum P5 and further studies are on-going for best cell growth and PHB yield.     

Effect of copper on the growth of shrimps Litopenaeus vannamei: water parameters and copper budget in a recirculating system

Shrimps( Litopenaeus vannamei) were intensively cultured in a recirculating aquaculture system for 98 days to investigate effects of 0.3 mg/L Cu on its performance, Cu budget, and Cu distribution. Shrimps in Cu-treated systems had greater mean final weight(11.10 vs 10.50 g), body length(107.23 vs 106.42 mm), survival rate(67.80% vs 66.40%), and yield(6.42 vs 5.99 kg/m 3), and lower feed conversion ratio(1.20 vs 1.29) than those in control systems but the differences were not significant. Vibrio numbers remained stable(10 4 –10 6 colony forming units/mL) in the rearing tanks of both control and treated systems. Total ammonium-N, nitrite-N, nitrate-N, pH, chemical oxygen demand, 5-day carbonaceous biochemical oxygen demand, and total suspended solids were similar in controls and treatments. Dissolved Cu concentration in the treated systems decreased from 0.284 to 0.089 mg/L while in the control systems it increased from 0.006 2 to 0.018 mg/L. The main sources of Cu in the treated systems were the artificially added component(75.7% of total input), shrimp feed(21.0%), water(2.06%), and shrimp biomass(1.22%). The major outputs of Cu occurred via the mechanical filter(41.7%), water renewal(15.6%), and draining of the sediment trap(15.1%). The foam fractionator removed only 0.69% of total Cu input. Harvested shrimp biomass accounted for 11.68% of Cu input. The Cu concentration of shrimps in the Cu-treated systems(30.70 mg/kg wet weight) was significantly higher than that in control systems(22.02 mg/kg). Both were below the maximum permissible concentration(50 mg/kg) for Cu in seafood for human consumption in China. Therefore, recirculating systems can be used for commercial on-growing of Litopenaeus vannamei without loss of shrimp quality, even in water polluted by 0.30 mg/L Cu. The mechanical filter is the main route for Cu removal.     

Genetic diversity analysis with ISSR PCR on green algae <Emphasis Type="Italic">Chlorella vulgaris</Emphasis> and <Emphasis Type="Italic">Chlorella pyrenoidosa</Emphasis>

In the present study, genetic polymorphism and diversity in unicellular clones of Chlorella vulgaris Beijerinck and Chlorella pyrenoidosa Chick were studied with Inter Simple Sequence Repeats PCR （ISSR PCR）. Samples including four clones of C. vulgaris and three clones of C. pyrenoidosa were purified by single-clone-choice method. For four C. vulgaris unicellular clones, the total number of the bands scored for 18 primers was 298; and the number of the polymorphic bands was 118, of which 39.6% were polymorphic. The size of PCR products ranged from 200 to 2 500 bp. The total number of bands scored for 18 primers, the number of polymorphic bands and the percentage of three C. pyrenoidosa unicellular clones was 194.83 and 30.8%, respectively. POPGENE analysis show that the average Nei genetic diversity （h^＊） and Shannon index of diversity （I^＊） in the four C. vulgaris unicellular clones was 0.2181 and 0.3208, respectively, which is slightly higher than those of the three C. pyrenoidosa unicellular clones （0.190 3 and 0.274 8）, which agreed with the percentage of polymorphic bands in the mixed samples of the two species. The results suggest that ISSR is a useful method to Chlorella for intra-species genetic analysis.     

一种户外开放式微藻培养体系优化

【目的】建立可用于大规模培养的户外开放式微藻培养体系。【方法】设计一种微藻平面开放浅层培养(Flat,open and shallow,FOS,简称浮法)体系,以小球藻(Chlorella sorokiniana)为模式藻种,探讨温度、光照、pH值和培养基营养成分等因素在该培养体系中对小球藻生长的影响,开展900 L体系的户外培养试验。【结果】浮法体系主要由塑料袋和垫板组成,在该培养体系中的小球藻在20~40℃范围内均可生长,最适生长温度为30~35℃。小球藻生长有明显的照度依赖性,无光时几不生长,随着照度升高,生长速度加快,在较高的照度下表现有光饱和现象。以TAP为基础培养基,在碳源或氮源缺乏时小球藻几不生长。pH 7.5左右有利于小球藻生长。900 L体系的户外培养试验的生物量(干物质得率)为0.15 g/(L·d)。【结论】在此新型户外平面开放浅层微藻培养体系中,温度、照度、pH值及营养成分等对小球藻的生长均有不同程度的影响。该体系有成本低、操作简便、容易控制、自然资源利用率高等特点,有大面积推广潜力。     

Genetic transformation of marine cyanobacterium Synechococcus sp. CC9311 (Cyanophyceae) by electroporation

Synechococcus sp.CC9311 is a marine cyanobacterium characterized by type IV chromatic acclimation(CA).A genetic transformation system was developed as a first step to elucidate the molecular mechanism of CA.The results show that Synechococcus sp.CC9311 cells were sensitive to four commonly used antibiotics:ampicillin,kanamycin,spectinomycin,and chloramphenicol.An integrative plasmid to disrupt the putative phycoerythrin lyase gene mpeV,using a kanamycin resistance gene as selectable marker,was constructed by recombinant polymerase chain reaction.The plasmid was then transformed into Synechococcus sp.CC9311 via electroporation.High transformation efficiency was achieved at a field strength of 2 kV/cm.DNA analysis showed that mpeV was fully disrupted following challenge of the transformants with a high concentration of kanamycin.In addition,the transformants that displayed poor growth on agar SN medium could be successfully plated on agarose SN medium.     

Effects of nutrients and light intensity on the growth and biochemical composition of a marine microalga Odontella aurita

Algal biotechnology has advanced greatly in the past three decades. Many microalgae are now cultivated to produce bioactive substances. Odontella aurita is a marine diatom industrially cultured in outdoor open ponds and used for human nutrition. For the first time, we have systematically investigated the effects of culture conditions in cylindrical glass columns and fiat-plate photobioreactors, including nutrients （nitrogen, phosphorus, silicon, and sulfur）, light intensity and light path, on O. aurita cell growth and biochemical composition （protein, carbohydrate, β-1,3-glucan, lipids, and ash）. The optimal medium for photoautotrophic cultivation of O. aurita contained 17.65 mmol/L nitrogen, 1.09 mmol/L phosphorus, 0.42 mmol/L silicon, and 24.51 mmol/L sulfur, yielding a maximum biomass production of 6.1-6.8 g/L and 6.7-7.8 g/L under low and high light, respectively. Scale-up experiments were conducted with fiat-plate photobioreactors using different light-paths, indicating that a short light path was more suitable for biomass production of O. aurita. Analyses of biochemical composition showed that protein content decreased while carbohydrate （mainly composed of 15-1,3-glucan） increased remarkably to about 50% of dry weight during the entire culture period. The highest lipid content （19.7% of dry weight） was obtained under 0.11 mmol/L silicon and high light conditions at harvest time. Fatty acid profiles revealed that 80% were Cx4, C~6, and C20, while arachidonic acid and eicosapentaenoic acid （EPA） accounted for 1.6%-5.6% and 9%-20% of total fatty acids, respectively. High biomass production and characteristic biochemical composition profiles make O. aurita a promising microalga for the production ofbioactive components, such as EPA and D-1,3-glucan.     

Genetic transformation of <Emphasis Type="Italic">Nannochloropsis oculata</Emphasis> with a bacterial phleomycin resistance gene as dominant selective marker

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2×10⁷ cells mL^?1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL^?1 zeocin. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.     

Study of phytoplankton community in the channel from Dongting Lake to Changjiang River, China

1 INTRODUCTION Dongting Lake (28°18'–29°45'N, 111°35'– 113°18'E) is the second largest freshwater lake in China, and directly connected with the Changjiang River (Yangtse River) through the channel. The sur- face area was about 2 740 km2 (at the low     

Phytoplanktonic desmids community in Donghu Lake, Wuhan,China

1 INTRODUCTION Donghu Lake (30o33′N, 114o23′E) is a shallow medium sized freshwater lake in Wuhan City, Hubei Province, China and about 5 kilometers away from the Changjiang (Yangtze) River. Before the 1960s, the water surface area was about 27.8 km2. The lake water was clear and the pH ranged from 6 to 7. The shallow area of the lake abounded with macrophytes. The phytoplanktonic and peripheral desmids were very abundant also in this lake. In the late 1960s, Donghu Lake with tota…     

The community structure and seasonal dynamics of plankton in Bange Lake,northern Tibet,China

The seasonal variations in biomass, abundance, and species composition of plankton in relation to hydrography were studied in the saline Bange Lake, northern Tibet, China. Sampling was carried out between one to three times per month from May 2001 to July 2002. Salinity ranged from 14 to 146. The air and water temperature exhibited a clear seasonal pattern, and mean annual temperatures were approximately 4.8°C and 7.3°C, respectively. The lowest water temperature occurred in winter from December to March at-2°C and the highest in June and July at 17.7°C. Forty-one phytoplankton taxa, 21 zooplankton, and 5 benthic or facultative zooplankton were identifi ed. The predominant phytoplankton species were Gloeothece linearis, Oscillatoria tenuis, Gloeocapsa punctata, Ctenocladus circinnatus, Dunaliella salina, and Spirulina major. The predominant zooplankton species included H olophrya actra, Brachionus plicatilis, Daphniopsis tibetana, Cletocamptus dertersi, and A rctodiaptomus salinus. The mean annual total phytoplankton density and biomass for the entire lake were 4.52×10~7 cells/L and 1.60 mg/L, respectively. The annual mean zooplankton abundance was 52, 162, 322, and 57, 144 ind./L, in the three sublakes. The annual mean total zooplankton biomass in Lakes 1–3 was 1.23, 9.98, and 2.13 mg/L, respectively. The annual mean tychoplankton abundances in Bg1, 2, and 3 were 47, 67, and 654 ind./L. The annual mean tychoplankton biomass was 2.36, 0.16, and 2.03 mg/L, respectively. The zooplankton biomass(including tychoplankton) in the lake was 9.11 mg/L. The total number of plankton species in the salt lake was signifi cantly negatively correlated with salinity.     

Bioreactor technology for production of valuable algal products

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

Optimization of medium and cultivation conditions for enhanced exopolysaccharide yield by marine Cyanothece sp. 113

Cyanothece sp. 113 is a unicellular, aerobic, diazotrophic and photosynthetic marine cyanobacterium. The optimal medium for exopolysaccharide yield by the strain was 70.0 g/L of NaCl, and 0.9 g/L of MgSO4 based on the modified F/2 medium for cultivation of marine algae. The optimal cultivation condition for exopolysaccharide yield by this cyanobacterial strain was 29°C, aeration, and continuous illumination at 86.0 μE/M2/S. Under the optimal conditions, over 18.4 g/L of exopolysaccharide was produced within 12 days. This was so far the highest exopolysaccharide yield produced with strains of Cyanothece sp. obtained.     

Spatiotemporal distribution of protozooplankton and copepod nauplii in relation to the occurrence of giant jellyfish in the Yellow Sea

The occurrence of the giant jellyfish, Nemopilema nomurai, has been a frequent phenomenon in the Yellow Sea. However, the relationship between the giant jellyfish and protozoa, in particular ciliates, remains largely unknown. We investigated the distribution of nanoflagellates, ciliates, Noctiluca scintillans, and copepod nauplii along the transect 33~N in the Yellow Sea in June and August, 2012, during an occurrence of the giant jellyfish, and in October of that year when the jellyfish was absent. The organisms studied were mainly concentrated in the surface waters in summer, while in autumn they were evenly distributed in the water column. Nanoflagellate, ciliate, and copepod nauplii biomasses increased from early June to August along with jellyfish growth, the first two decreased in October, while N. scintillans biomass peaked in early June to 3 571 pg C/L and decreased in August and October. In summer, ciliate biomass greatly exceeded that of copepod nauplii （4.61-15.04 ~tg C/L vs. 0.34-0.89 pg C/L）. Ciliate production was even more important than biomass, ranging from 6.59 to 34.19 ~tg C/（L.d） in summer. Our data suggest a tight and positive association among the nano-, micro-, and meso-zooplankton in the study area. Statistical analysis revealed that the abundance and total production of ciliate as well as loricate ciliate biomass were positively correlated with giant jellyfish biomass, indicating a possible predator-prey relationship between ciliates and giant jellyfish. This is in contrast to a previous study, which reported a significant reduction in ciliate standing crops due to the mass occurrence ofN. nomurai in summer. Our study indicates that, with its high biomass and, in particular, high production ciliates might support the mass occurrence of giant jellyfish.     

<Emphasis Type="Italic">GST</Emphasis> (<Emphasis Type="Italic">phi</Emphasis>) gene from Macrophyte <Emphasis Type="Italic">Lemna minor</Emphasis> is involved in cadmium exposure responses

Reactive oxygen species (ROS) scavengers, including ascorbate peroxidase, superoxide dismutase, catalase and peroxidase, are the most commonly used biomarkers in assessing an organisms’ response to many biotic and abiotic stresses. In this study, we cloned an 866 bp GST (phi) gene in Lemna minor and investigated its characteristics, expression and enzymatic activities under 75 μmol/L cadmium concentrations in comparison with other ROS scavengers. GST (phi) gene expression patterns were similar to those of other scavengers of ROS. This suggests that GST (phi) might be involved in responding to heavy metal (cadmium) stress and that its expression level could be used as a bio-indicator in monitoring cadmium pollution.     

Effects of DDT and BHC on amino acid content and its varieties inChlorella vulgaris Beij. andCladophora sp.

InChlorella vulgaris Beij. andCladophora sp. treated with different concentrations of r-BHC and p, p-DDT, the protein and free amino acid content in both were higher than those in the controls, and the free amino acid content was even higher than the protein amino acid content.     

Size-fractionated Chlorophyll <Emphasis Type="Italic">a</Emphasis> biomass in the northern South China Sea in summer 2014

Spatial distribution of phaeopigment and size-fractionated chlorophyll a (Chl a) concentrations were examined in relation to hydrographic conditions in the northern South China Sea (NSCS) during a survey from 20 August to 12 September, 2014. The total Chl a concentration varied from 0.006 to 1.488 µg/L with a mean value of 0.259±0.247 (mean±standard deviation) µg/L. Chl a concentration was generally higher in shallow water (<200 m) than in deep water (>200 m), with mean values of 0.364±0.311 µg/L and 0.206±0.192 µg/L respectively. Vertically, the maximum total Chl a concentration appeared at depths of 30–50 m and gradually decreased below 100 m. The size-fractionated Chl a concentrations of grid stations and time-series stations (SEATS and J4) were determined, with values of pico- (0.7–2 µm), nano- (2–20 µm) and micro-plankton (20–200 µm) ranging from 0.001–0.287 (0.093±0.071 µg/L), 0.004–1.149 (0.148±0.192 µg/L) and 0.001–0.208 (0.023±0.036 µg/L), respectively. Phaeopigment concentrations were determined at specific depths at ten stations, except for at station A9, and varied from 0.007 to 0.572 (0.127±0.164) µg/L. Nano-and pico-plankton were the major contributors to total phytoplankton biomass, accounting for 50.99%±15.01% and 39.30%±15.41%, respectively, whereas microplankton only accounted for 9.39%±8.66%. The results indicate that the contributions of microplankton to total Chl a biomass were less important than picoplankton or nanoplankton in the surveyed NSCS. Different sized-Chl a had similar spatial patterns, with peak values all observed in subsurface waters (30–50 m). The summer monsoon, Kuroshio waters, Zhujiang (Pearl) River plume, and hydrological conditions are speculated to be the factors controlling the abundance and spatial heterogeneity of Chl a biomass in the NSCS.     

Study on the phytoplankton · in a large reservoir

A comprehensive study on the community structure and function of the phytoplankton in Taipinghu Reservoir, the largest reservoir (9400 ha) in Anhui Province, China, was carried out during 1985–1986. A total of 175 species of algae belonging to 8 phyla and 87 genera was noted. The composition of phytoplankton was dominated by species of Chlorophyta, Cyanophyta and Bacillariophyta. The species number, cell density and biomass of the three groups were respectively 88%, 86.5% and 78.9% of the total phytoplankton. The weighted annual average biomass was 1.52 mg/L and cell density was 1.43×10⁶ ind/L. The growth maximum was observed in summer. Diatoms were abundant in the region adjoining a river. A large number of flagellated algae such asEuglena andChromulina occurred in the artificial fish culture bay. It can be inferred from the algal composition and total nitrogen concentration (1.48 mg/L) that this reservoir is a mesotrophic water body that had undergone slight natural eutrophication. Presented are an equation for the relation between cell density and biomass of algae and a newly developed method based on the energy flow principle in ecology for theoretically estimating algal fishery potential. The algae volumes of 128 species were measured for calculating the biomass.     

Effect of carbon source on growth and lipid accumulation in <Emphasis Type="Italic">Chlorella sorokiniana</Emphasis> GXNN01

Heterotrophic culture of microalgae to develop methods of increasing biomass productivity and storage lipids has brought new insight to commercial biodiesel production. To understand the relationship between heterotrophy and lipid production, the effects of carbon sources on the growth and lipid accumulation of Chlorella sorokiniana GXNN01 was studied. The alga exhibited an increased growth rate in response to the addition of carbon sources, which reached the stationary phase after 48 h at 30°C. In addition, glucose and NaAc had a significant effect on the lipid accumulation during the early-stationary phase. Specifically, the lipid content was 0.237±0.026 g g⁻¹ cell dry weight and 0.272±0.041 g L⁻¹ when glucose was used as the carbon source, whereas the lipid content reached 0.287±0.018 g g⁻¹ cell dry weight and 0.288±0.008 g L⁻¹ when NaAc was used as the carbon source. The neutral lipid content was found to first decrease and then increase over time during the growth phase. A glucose concentration of 20 mmol L⁻¹ gave the maximal lipid yield and the optimum harvest time was the early-stationary phase.     

Phytoplankton Community Dynamics in West Lake After Drawing Water from the Qiantang River

1INTRODUCTIONTheWestLake ( 30°1 5′N ,1 2 0°1 6′E)isasmallshallowlakeinthewesternpartofHangzhouCityinthesoutheasterncoastalareaofChina.Thelakeissur roundedonthreesidesbyhills ;andisfamousforitspicturesquesceneryandinterfusionofhillsandwaterbodies.Thesurfaceareaisabout 5 .66km2 andmeandepthis 1 .8m .Beforethe 1 95 0s ,thelakewaterwascleanandmacrophytesgrewveryluxuriantlyinthelit toralareasofthelake .In 1 95 2 ,WestLakewasdredgedand 70 0× 1 0 4m3 mudwasremovedfromthebottomofthelake …