Intergeneric conjugation in holomycin-producing marine Streptomyces sp strain M095

Marine Streptomyces are potential candidates for novel natural products and industrial catalysts. In order to set up biosynthesis approach for a holomycin-producing strain M095 isotated from Jiaozhou Bay, China, a genetic transformation system was established using intergeneric conjugation. The plasmid pIJ8600 consists of an origin of replication for Escherichia coli, a phage integrase directing efficient site-specific integration in bacterial chromosome, thiostrepton-induced promoter and an attP sequence. Using E. coli ET12567 (pUZ8002) carrying pIJ8600 as a conjugal donor, while it was mated with strain M095, pIJ8600 was mobilized to the recipient and the transferred DNA was also integrated into the recipient chromosome. The frequency of exconjugants was 1.9 +/- 0.13 x 10(-4) per recipient cell. Analysis of eight exconjugants showed pIJ8600 was stable integrated at a single chromosomal site (attB) of the Streptomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of growth and antimicrobial activity analysis indicated that the integration of pIJ8600 did not seem to affect the biosynthesis of antibiotics or other essential amino acids. To demonstrate the feasibility of above gene transfer system, the allophycocyanin gene (apc) from cyanobacterium Anacystis nidulans UTEX625 was expressed in strain M095, and the results indicated heterologous allophycocyanin could be expressed and folded effectively. (c) 2006 Elsevier GmbH. All rights reserved.

New sphingolipids with a previously unreported 9-Methyl-C-20-sphingosine moiety from a marine algous endophytic fungus Aspergillus niger EN-13

Asperamides A (1) and B (2), a sphingolipid and their corresponding glycosphingolipid possessing a hitherto unreported 9-methyl-C-20-sphingosine moiety, were characterized from the culture extract of Aspergillus niger EN-13, an endophytic fungus isolated from marine brown alga Colpomenia sinuosa. The structures were elucidated by spectroscopic and chemical methods as (2S,2'R,3R,3'E,4E,8E)-N-(2'-hydroxy-3'-hexadecenoyl)-9-methyl-4,8-icosadien-1,3-diol (1) and 1-O-beta-D-glucopyranosyl-(2S,2'R,3R,3'E,4E,8E)-N-(2'-hydroxy-3'-hexadecenoyl)-9-methyl-4,8-icosadien-1,3-diol (2). In the antifungal assay, asperamide A (1) displayed moderate activity against Candida albicans.

Triterpenoids from the mangrove plant Hibiscus tiliaceus

Three new triterpenoids with the rarely occurring nigrum skeleton, namely (20E)-22-hydroxynigrum-20-en-3-one (1), 21 beta-hydroxynigrum-22(29)-en-3-one (2), and 21 alpha-hydroxynigrum-22(29)-en-3-one (3), were isolated from the mangrove plant Hibiscus tiliaceus. Additionally, five known triterpenoids including friedelin (4),12-oleanen-3 beta-ol (5), 3 beta-hydroxy-12-oleanen-28-oic acid (6),20(29)-lupen-3 beta,28-diol (7). and cucurbita-5,23-dien-3 beta,25-diol (8) were also isolated and identified. The latter structures were elucidated by a detailed NMR and MS analyses, as well as by comparison with reported literature data.

attB site disruption in marine Actinomyces sp M048 via DNA transformation of a site-specific integration vector

An efficient conjugation method has been developed for the marine Actinomyces sp. isolate M048 to facilitate the genetic manipulation of the chandrananimycin biosynthesis gene cluster. A phi C31-derived integration vector pIJ8600 containing oriT and attP fragments was introduced into strain M048 by bi-parental conjugation from Escherichia coli ET12567 to strain M048. Transformation efficiency was (6.38 +/- 0.41) x 10(-5) exconjugants per recipient spore. Analysis of eight exconjugants showed that the plasmid pIJ8600 was stably integrated at a single chromosomal site (attB) of the Actinomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of antimicrobial activity analysis indicated that the insertion of plasmid pIJ8600 seemed to affect the biosynthesis of antibiotics that could strongly inhibit the growth of E. coli and Mucor miehei (Tu284). HPLC-MS analysis of the extracts indicated that disruption of the attB site resulted in the complete abolition of chandrananimycin A-C production, proving the identity of the gene cluster. Instead of chandrananimycins, two bafilomycins were produced through disruption of the attB site from the chromosomal DNA of marine Actinomyces sp. M048.

Propagators and ridge jumps in a back-arc basin, the West Philippine Basin

We explore the tectono-magmatic processes in the western West Philippine Basin, Philippine Sea Plate, using bathymetric data acquired in 2003 and 2004. The northwestern part of the basin formed through a series of northwestward propagating rifts. We identify at least five sequences of propagating rifts, probably triggered by mantle flow away from the mantle thermal anomaly that is responsible for the origin of the Benham and Urdenata plateaus. Gravitational forces caused by along-axis topographic gradient and a similar to 30 degrees ridge reorientation appear to also be driving the rift propagations. The along-axis mantle flow appears to be reduced and deflected along the Luzon-Okinawa fracture zone, because the spreading system remained stable west of this major fault zone. North-east of the Benham plateau, a left-lateral fracture zone has turned into a NE-SW-trending spreading axis. As a result, a microplate developed at the triple junction.

A semi-analytical theory of coastal upwelling and jet

The general forms of the conservation of momentum, temperature and potential vorticity of coastal ocean are obtained in the x-z plane for the nonlinear ocean circulation of Boussinesq fluid, and a elliptic type partial differential equations of second order are derived. Solution of the partial differential equations are obtained under the conditions that the fluid moves along the topography. The numerical results show that there exist both upwelling and downwelling along coastline that mainly depends on the large scale ocean condition. Numerically results of the upwelling (downwelling), coastal jet and temperature front zone are favorable to the observations.

Feeding and egg production of the planktonic copepod Calanus sinicus in spring and autumn in the Yellow Sea, China

Shipboard incubations were conducted in spring (April) and autumn (October/November) 2006 to measure the feeding and egg production rates (EPR) of Calanus sinicus in the Yellow Sea, China. The ingestion rate (2.08-11.46 and 0.26-3.70 mu g C female(-1) day(-1) in spring and autumn, respectively) was positively correlated with microplankton carbon concentrations. In the northern part of the Yellow Sea, feeding on microplankton easily covers the respiratory and production requirements, whereas in the southern part in spring and in the frontal zone in autumn, C. sinicus must ingest alternative food sources. Low ingestion rates, no egg production and the dominance of the fifth copepodite (CV) stage indicated that C. sinicus was in quiescence inside the Yellow Sea Cold Bottom Water (YSCBW) area in autumn. Calanus sinicus ingested ciliates preferentially over other components of the microplankton. The EPR (0.16-12.6 eggs female(-1) day(-1) in spring and 11.4 eggs female(-1) day(-1) at only one station in autumn) increased with ciliate standing stock. Gross growth efficiency (GGE) was 13.4% (3-39%) in spring, which was correlated with the proportion of ciliates in the diet. These results indicate that ciliates have higher nutrient quality than other food items, but the low GGE indicates that the diet of C. sinicus is nutritionally incomplete.

Typhoon effects on DOC dynamics in a phosphate-limited reservoir

To explore typhoon effects on dissolved organic carbon (DOC) dynamics, field investigations (tributary and dam site) and laboratory experiments (bioassay and DOC consumption) were conducted in a subtropical reservoir. A tributary survey indicated that after typhoon disruption, upstream areas were the sources of phosphate (P) but not DOC for the dam site located downstream. Bioassay experiments verified P-limitation on bacteria and phytoplankton during summer stratification, and bacteria showed a faster response than algae to added P. Experiments indicated that DOC consumption was determined by the availability of P. The 4 yr typhoon period (June-September) data of the dam site denoted that DOC concentration (27 to 270 mu M C) and its rate of change (-13 to 24 mu M C d(-1)) varied more dramatically in the weak (2006 and 2007) than in the strong (2004 and 2005) typhoon years. The negative correlation of DOC with the ratio of bacterial production (BP) to primary production (PP) in the euphotic zone (0 to 10 m) signified the interactive effects of auto- and heterotrophic processes on DOC variation. In the aphotic zone, the variation of DOC could be ascribed to the change of BP, which showed a positive correlation with P concentrations. This study documents that DOC concentration in the studied system varied at multiple time scales. Such variation can be explained by the decoupling between BP and PP, which is believed to be a function of the limiting nutrient's availability. More importantly, this study suggests that the P supply introduced by strong typhoons might have substantiated a tighter coupling between BP and PP, so that the amplitude of DOC oscillation during the summer period was effectively reduced.

Nutrient characteristics in the Yangtze River Estuary and the adjacent East China Sea before and after impoundment of the Three Gorges Dam

From November 2002 to 2006, five cruises were undertaken in the Yangtze River Estuary and the adjacent East China Sea to compare the nutrient concentrations, ratios and potential nutrient limitation of phytoplankton growth before and after impoundment (June 2003) of the Three Gorges Dam (TGD). Concentrations of dissolved inorganic nitrogen (DIN), soluble reactive phosphorus (SRP) and total nitrogen (TN) exhibited an increasing trend from 2002 to 2006. In contrast, total phosphorus (TP) concentration exhibited a decreasing trend. The mean concentrations of DIN, SRP, and TN in the total study area increased from 21.4 mu M, 0.9 mu M, and 41.8 mu M in 2002 to 37.5 mu M, 1.3 mu M. and 82.2 mu M in 2006, respectively. while TP decreased from 2.1 mu M to 1.7 mu M. The concentration of dissolved reactive silica (DRSi) had no major fluctuations and the differences were not significant. The mean concentration of DRSi in the total study area ranged from 52.5 to 92.3 mu M. The Si:N ratio decreased significantly from 2.7 in 2002 to 1.3 in 2006, while TN: TP ratio increased from 22.1 to 80.3. The area of potential P limitation of phytoplankton growth expanded after 2003 and potential Si limitation appeared in 2005 and 2006. Potential P limitation mainly occurred in an area of salinity less than 30 after 2003, while potential Si limitation occurred where the salinity was greater than 30. By comparison with historical data, the concentrations of nitrate and SRP in this upper estuary during November 1980-2006 increased obviously after impoundment of TGD but DRSi decreased. Meanwhile, the ratios of N:P, Si:N and Si:P decreased obviously. (c) 2009 Elsevier B.V. All rights reserved.

Effects of different diets on the reproduction and naupliar development of the copepod Acartia bifilosa

The influence of diatoms on the reproduction and naupliar development of Acartia bifilosa was investigated under laboratory conditions, comparing initial in situ values and laboratory-food treatments. Egg production by A. bifilosa was significantly reduced by one diatom diet (Phaeodactylum tricornutum: Pt) and by two non-diatom diets (Platymonas subordiformis: Ps and Nannochloropsis oculata: No). It was less affected by the other diatom diet (Skeletonema costatum: Sc) or by two mixed-food treatments (D-mix and DG-mix), composed of two diatoms (Pt, Sc) and four species (Pt, Sc, Ps and No), respectively. The negative effect of Pt was eliminated when adult copepods were offered mixed-food diets. There were no significant differences between the hatching success values observed in filtered seawater and in algal exudates, indicating that diatoms did not produce active dissolved toxic substances under the different food concentrations tested. The mortality rate of nauplii was higher with Pt than the other diets, suggesting that this diatom species had a negative effect on egg production, hatching success and naupliar survival simultaneously. Compared to other diets, No and Pt were not beneficial food sources for reproduction and for female and larval survival. (c) 2007 Elsevier B.V. All rights reserved.

Feeding habits of Calanus sinicus (Crustacea : Copepoda) during spring and autumn in the Bohai Sea studied with the herbivore index

Pigment ingestion rate (PIR) and egg production rate (EPR) of the dominant copepod Calanus sinicus, as well as chlorophyll-a concentration and phytoplankton assemblages were measured in the Bohai Sea, North China in June 1997, October 1998 and May 1999. A herbivore index (H) was also calculated as the carbon specific ratio of PIR and EPR, in order to investigate its feeding habits in the spring and autumn phytoplankton bloom respectively. On average, chlorophyll-a concentration was relatively similar (1-1.34 mg m(-3)) in the three cruises, but PIR was quite different. It was 3.24 mu g C female(-1) d(-1) in October, equivalent to one half of the PIR for June and one third of the PIR for May. Average EPR was highest in May, and quite similar during the other two months. According to H values, herbivorous feeding contributed 100% of the egg production of C. sinicus in June, 82.5% in May, but only 47.8% in October. It is possible that omnivorous feeding of C. sinicus in October was induced by a prevalence of large-sized diatoms and sufficient non-phytoplankton food resources during the autumn bloom period.

Community structure of macrobenthos in coastal water off Rushan, southern Shandong Peninsula, and the relationships with environmental factors

To understand the present actuality of the marine ecosystem in the southern coastal water region of the Shandong Peninsula and the impact of the global change and the human activities to the marine ecosystem of the region, the macrobenthic community structure was researched based on data from 26 sampling stations carried out on four seasonal cruises from December 2006 to November 2007. The data was analyzed using PRIMER 6.0 and SPSS 15.0 software packages. The results showed that 236 macrobenthic species in total were collected from the research region by the field works. Most of the species belong to Polychaeta (76 species), Mollusca (75) and Crustacea (60). Of which, 33 species were common species by the four cruises. The dominant species were different among the four seasons, however, the polychaete species Nephtys oligobranchia and Sternaspis scutata were always dominant in the four seasons. The abundances and biomasses of the macrobenthos from the research region were variable in tire four seasons. The results of CLUSTER and MDS analysis showed that the similarities of macrobenthic structures among the stations were low, most of the similarities were at about 40% of similarity values, only that of two stations were up to 60%. In accordance with the similarity values of the macrobenthic structures, the 26 stations were clustered as six groups at arbitrary similarity level of 30%. The ABC curve indicated that the marcofauna communities in the research region had riot been disturbed distinctly. The results of BIOENV and BVSTEP (Spearman) analysis implied that the concentrations of organic matter in bottom water and heavy metal copper in sediment, water depth and temperature of bottom were the most significant environmental factors to affect the macrobentic community.

Mantle-derived gaseous components in ore-forming fluids of the Xiangshan uranium

The Xiangshan U deposit, the largest hydrothermal U deposit in China, is hosted in late Jurassic felsic volcanic rocks although the U mineralization post dates the volcanics by at least 20 Ma. The mineralization coincides with intrusion of local mantle-derived mafic dykes formed during Cretaceous crustal extension in South China. Ore-forming fluids are rich in CO2, and U in the fluid is thought to have been dissolved in the form of UO2 (CO3)22− and UO2 (CO3) 34− complexes. This paper provides He and Ar isotope data of fluid inclusions in pyrites and C isotope data of calcites associated with U mineralization (pitchblende) in the Xiangshan U deposit. He isotopic compositions range between 0.1 and 2.0Ra (where Ra is the 3He/4He ratio of air=1.39×10−6) and correlates with 40Ar/36Ar; although there is potential for significant 3He production via 6Li(n,α)3H(β)3He reactions in a U deposit (due to abundant neutrons), nucleogenic production cannot account for either the 3He concentration in these fluids, nor the correlations between He and Ar isotopic compositions. It is more likely that the high 3He/4He ratios represent trapped mantle-derived gases. A mantle origin for the volatiles of Xiangshan is consistent with the δ13C values of calcites, which vary from −3.5‰ to −7.7‰, overlapping the range of mantle CO2. The He, Ar and CO2 characteristics of the ore-forming fluids responsible for the deposit are consistent with mixing between 3He- and CO2-rich mantle-derived fluids and CO2-poor meteoric fluids. These fluids were likely produced during Cretaceous extension and dyke intrusion which permitted mantle-derived CO2 to migrate upward and remobilize U from the acid volcanic source rocks, resulting in the formation of the U deposit. Subsequent decay of U within the fluid inclusions has reduced the 3He/4He ratio, and variations in U/3He result in the range in 3He/4He observed with U/3He ratios in the range 5–17×103 likely corresponding to U concentrations in the fluids b0.2 ppm.