Molecular cloning, expression of a peroxiredoxin gene in Chinese shrimp Fenneropenaeus chinensis and the antioxidant activity of its recombinant protein

Peroxiredoxin (Prx) is known to be an antioxidant protein that protects the organisms against various oxidative stresses and functions in intracellular signal transduction. A Prx gene was firstly isolated in the crustacean, Chinese shrimp Fenneropenaeus chinensis. The full-length cDNA consists of 942 bp with a 594 bp open reading frame, encoding 198 amino acids. The molecular mass of the deduced amino acid is 22041.17 Da with an estimated pI of 5.17. Sequence comparison showed that Prx of F. chinensis shares 76%, 73% and 72% identity with that of Aedes aegypti, Branchiostoma belcheri tsingtaunese and Drosophila melanogaster, respectively. Northern blot analysis revealed the presence of Prx transcripts of F chinensis in all tissues examined. Real-time PCR analysis indicated that the Prx showed different expression profiles in shrimp hemocytes and hepatopancreas after artificial infection with Vibrio anguillarum. In addition, a fusion protein containing Prx was produced in vitro. LC-ESI-MS analysis showed that four peptide fragments of the recombinant protein were identical to the corresponding sequence of F. chinensis Prx. And the purified recombinant proteins were shown to reduce H2O2 in the presence of dithiothreitol. (c) 2007 Elsevier Ltd. All rights reserved.

Analysis of the expression and antioxidative property of a peroxiredoxin 6 from Scophthalmus maximus

Peroxiredoxins (Prxs) are a group of antioxidant proteins that protect cells from oxidative damage caused by various peroxides. To date, six different isoforms of peroxiredoxin (Prx1 to Prx6) have been identified, of which, Prx6 belongs to the 1-Cys Prx subfamily. Although Prx6 of several fish species have been reported at sequence level, there are very few documented studies on the potential function of fish Prx6. In this report, we describe the identification and analysis of a Prx6 homologue, SmPrx6, from turbot Scophthalmus maximus. The full length cDNA of SmPrx6 contains a 5'- untranslated region (UTR) of 60 bp, an open reading frame of 666 bp, and a 3'-UTR of 244 bp. The deduced amino acid sequence of SmPrx6 shares 81-87% overall identities with known fish Prx6. In silico analysis identified in SmPrx6 a conserved Prx6 catalytic motif, PVCTTE, and the catalytic triads putatively involved in peroxidase and phospholipase A2 activities. Expression of SmPrx6 was detected in most fish organs, with the highest expression levels found in blood and heart and the lowest level in spleen. Experimental challenges with bacterial pathogens and poly(I:C) upregulated SmPrx6 expression in liver and spleen in a manner that is dependent on the challenging agent and the tissue type. Treatment of cultured primary hepatocytes with H2O2 enhanced SmPrx6 expression in a dose-dependent manner. Recombinant SmPrx6 expressed in and purified from Escherichia coli exhibited thiol-dependent antioxidant activity and could protect cultured hepatocytes from H2O2-induced oxidative damage. Taken together, these results indicate that SmPrx6 is a Prx6 homologue with antioxidative property and is likely to be involved in both cellular maintenance and protective response during host immune defense against bacterial infection. (C) 2010 Elsevier Ltd. All rights reserved.

Genetic transformation in Kappaphycus alvarezii using micro-particle bombardment: a potential strategy for germplasm improvement

This study investigated the delivery of a SV40 promoter driving lacZ gene into cells of Kappaphycus alvarezii using particle bombardment. Thallus pieces 0.5-0.8 mm in diameter and 1 cm in length were prepared as gene recipients. Bombardment parameters of 450 psi (rupture pressures) x 6 cm (particle travel distances), 650 psi x 6 cm, 1,100 psi x 6 cm and 1,100 psi x 9 cm were used. A significant increase in transformation efficiency from about 33% under the rupture pressure of 450 psi to 87% at 650 psi was observed in transformed thalli. Most of the positive cells appeared in epidermal cells bombarded at 450 psi, whereas positive signals were seen in both epidermal and medullary cells at 650 psi. No positive transient expression was detected at a bombardment of 1,100 psi, or in negative or blank controls. For the conditions tested, the best parameter was obtained at 650 psi at a distance of 6 cm. Thus, the strategy of taking vegetative thalli as recipients, using particle bombardment, and combining this with micro-propagation, together with developing an in vivo selectable marker, is a viable way to produce stable transformants, to eliminate chimeric expression, and to achieve transgenic breeding in K. alvarezii.

Proteomic analysis of differentially expressed proteins in lymphoid organ of Fenneropenaeus chinensis response to Vibrio anguillarum stimulation

To gain an insight into the function of shrimp lymphoid organ at protein level, we analyzed the proteome of lymphoid organ in healthy Chinese shrimp Fenneropenaeus chinensis (F. chinensis) through two-dimensional gel electrophoresis (2-DE) based proteomic approach. A total of 95 spots representing 75 protein entries were identified by liquid chromatography tandem mass spectrometry (LC-MS/MS) with both online and in-house database. According to Gene Ontology (GO) annotation of biological process, the identified proteins were classified into 13 categories. Among them, approximately 36% of proteins related to cytoskeleton are noticeable. Then, a comparative proteomic approach was employed to investigate the differentially expressed proteins in lymphoid organ of Vibrio anguillarum-challenged F. chinensis. At 24 h post-injection (hpi), 17 differentially expressed protein spots were successfully identified, including 4 up-regulated protein spots (represent 4 proteins: cathepsin L protein similar to squid CG16901-PC, protein kinase C and protein similar to T-complex Chaperonin 5 CG8439-PA), and 13 down-regulated protein spots (represent 9 proteins: actin, beta-actin, cytoplasmic actin CyII, alpha tubulin, beta tubulin, protein similar to proteasome delta, vacuolar ATP synthase subunit B, elongation factor 2, carboxypeptidase B). These data may help us to understand the function of lymphoid organ and the molecular immune mechanism of shrimp responsive to pathogen infection. (C) 2010 Elsevier Ltd. All rights reserved.

Relationship between differential retention of Escherichia coli and Enterococcus faecalis and variations in enzyme activity in the scallop Patinopecten yessoensis

Uptake of Escherichia coli and Enterococcus faecalis and variations of trypsin amylase activity acid phosphatase and superoxide dismutase in tissue of the scallop Patinopecten yessoensis were detected. The results showed that P. yessoensis accumulated E. faecalis in larger numbers and more rapidly than E. coli, both with the highest concentration in the digestive tract and lowest in hemolymph. Compared to E. coil, all scallops exposed to E. faecalis showed significantly higher trypsin and AMS activity. SOD activity in hemocytes and ACP activity in hemolymph was significantly higher in the treatments with 5 log(10)CFU/ml E. colt than with E. faecalis. But no significant differences in ACP activity of P. yessoensis exposed to a 3 log(10)CFU/ml inoculum of both bacteria were recorded. In conclusion, the mass retention of gut microflora in P. yessoensis is positively correlated with digestive enzymes activity and negatively correlated with ACP activity in the hemocyte. (C) 2010 Published by Elsevier Ltd.

Ulva and Enteromorpha (Ulvaceae, Chlorophyta) from two sides of the Yellow Sea: analysis of nuclear rDNA ITS and plastid rbcL sequence data

Ulvacean green seaweeds are common worldwide; they formed massive green tides in the Yellow Sea in recent years, which caused marine ecological problems as well as a social issue. We investigated two major genera of the Ulvaceae, Ulva and Enteromorpha, and collected the plastid rbcL and nuclear ITS sequences of specimens of the genera in two sides of the Yellow Sea and analyzed them. Phylogenetic trees of rbcL data show the occurrence of five species of Enteromorpha (E. compressa, E. flexuosa, E. intestinalis, E. linza and E. prolifera) and three species of Ulva (U. pertusa, U. rigida and U. ohnoi). However, we found U. ohnoi, which is known as a subtropical to tropical species, at two sites on Jeju Island, Korea. Four ribotypes in partial sequences of 5.8S rDNA and ITS2 from E. compressa were also found. Ribotype network analysis revealed that the common ribotype, occurring in China, Korea and Europe, is connected with ribotypes from Europe and China/Japan. Although samples of the same species were collected from both sides of the Yellow Sea, intraspecific genetic polymorphism of each species was low among samples collected worldwide.

Screening of genes related to ovary development in Chinese shrimp Fenneropenaeus chinensis by suppression subtractive hybridization

The ovary of triploid shrimp Fenneropenaeus chinensis was apparently impaired compared to that of the diploid shrimp at the same age. Therefore triploid shrimp ovary is possible to be taken as a model to understand the mechanism of ovary development of shrimp compared to that of the ovary of diploid shrimp at the same age. In the present study, a suppression subtractive hybridization (SSH) technique was applied to identify differentially expressed genes in the ovary between diploid and triploid shrimp. For the forward library (RNA from the ovary of triploid shrimp as the tester), 54 genes were identified. For the reverse library (RNA from the ovary of diploid shrimp as the tester), 16 genes were identified. The identified genes encoded proteins with multiple functions, including extracellular matrix components, cytoskeleton, cell growth and death, metabolism, genetic information processing, signal transduction/transport or immunity related proteins. Eleven differentially expressed genes were selected to be confirmed in the ovaries of triploid and diploid shrimp by semi-quantitative RT-PCR. Genes encoding spermatogonial stem-cell renewal factor, cytochrome c oxidase subunits I and II, clottable protein, antimicrobial peptide and transposase showed up-regulated expressions in the ovary of triploid shrimp. Genes encoding tubulin, cellular apoptosis susceptibility protein, farnesoic acid O-methyltransferase, thrombospondin and heat shock protein 90 genes showed higher expressions in the ovary of diploid shrimp. The differential expressions of the above genes are suggested to be related to the ovary development of shrimp. It will provide a new clue to uncover the molecular mechanisms underlying the ovarian development in penaeid shrimp. (C) 2010 Elsevier Inc. All rights reserved.

Identification and analysis of a Scophthalmus maximus ferritin that is regulated at transcription level by oxidative stress and bacterial infection

Ferritins are conserved Iron storage proteins that exist in most living organisms and play an essential role in Iron homeostasis. In this study, we reported the identification and analysis a ferritin M subunit, SmFerM, from turbot Scophthalmus maximus. The full length cDNA of SmFerM contains a 5'-untranslated region (UTR) of 232 bp, an open reading frame (ORF) of 531 bp, and a 3'-UTR of 196 bp The ORF encodes a putative protein of 176 amino acids, which shares extensive sequence identities with the M terrains of several fish species. In silico analysis identified in SmFerM both the ferroxidase center of mammalian H ferritins and the iron nucleation site of mammalian L ferritins. Quantitative real time reverse transcriptase-PCR analysis indicated that SmFerM expression was highest in muscle and lowest in heart and responded positively to experimental challenges with bacterial pathogens and poly(I center dot C) Exposure of cultured turbot hepatocytes to treatment of stress inducers (iron, copper, and H2O2) significantly upregulated the expression of SmFerM in a dose dependent manner. Iron chelating analysis showed that recombinant SmFerM purified from Escherichia coli exhibited apparent iron binding activity. These results suggest that SmFerM is a functional M ferritin and is likely to play a role in iron sequestration and protection against oxidative stress and microbial infection (C) 2010 Elsevier Inc All rights reserved

Surface sediment diatoms from the western Pacific marginal seas and their correlation to environmental variables

Diatom data of 192 surface sediment samples from the marginal seas in the western Pacific together with modern summer and winter sea surface temperature and salinity data were analyzed. The results of canonical correspondence analysis show that summer sea-surface salinity (SSS) is highly positively correlated with winter SSS and so is summer sea-surface temperature (SST) with winter SST. The correlations between SSSs and SSTs are less positively correlated, which may be due to interactions of regional current pattern and monsoon climate. The correlations between diatom species, sample sites and environmental variables concur with known diatom ecology and regional oceanographic characters. The results of forward selection of the environmental variables and associated Monte Carlo permutation tests of the statistical significance of each variable suggest that summer SSS and winter SST are the main environmental factors affecting the diatom distribution in the area and therefore preserved diatom data from down core could be used for reconstructions of summer SSS and winter SST in the region.

Quantitatively distinguishing sediments from the Yangtze River and the Yellow River using delta Eu-N-I REEs pound plot

Sediment samples were collected from the lower channel of the Yangtze River and the Yellow River and the contents of rare earth elements (REEs) were measured. In addition, some historical REEs data were collected from published literatures. Based on the delta Eu-N-I REEs pound plot, a clear boundary was found between the sediments from the two rivers. The boundary can be described as an orthogonal polynomial equation by ordinary linear regression with sediments from the Yangtze River located above the curve and sediments from the Yellow River located below the curve. To validate this method, the REEs contents of sediments collected from the estuaries of the Yangtze River and the Yellow River were measured. In addition, the REEs data of sediment Core 255 from the Yangtze River and Core YA01 from the Yellow River were collected. Results show that the samples from the Yangtze River estuary and Core 255 almost are above the curve and most samples from the Yellow River estuary and Core YA01 are below the curve in the delta Eu-N-I REEs pound plot. The plot and the regression equation can be used to distinguish sediments from the Yangtze River and the Yellow River intuitively and quantitatively, and to trace the sediment provenance of the eastern seas of China. The difference between the sediments from two rivers in the delta Eu-N-I REEs pound plot is caused by different mineral compositions and regional climate patterns of the source areas. The relationship between delta Eu-N and I REEs pound is changed little during the transport from the source area to the river, and from river to the sea. Thus the original information on mineral compositions and climate of the source area was preserved.

Formation and evolution of the modern warm current system in the East China Sea and the Yellow Sea since the last deglaciation

To reconstruct the formation and evolution process of the warm current system within the East China Sea (ECS) and the Yellow Sea (YS) since the last deglaciation, the paleoceangraphic records in core DGKS9603, core CSH1 and core YSDP102, which were retrieved from the mainstream of the Kuroshio Current (KC), the edge of the modern Tsushima Warm Current (TWC) and muddy region under cold waters accreted with the Yellow Sea Warm Current (YSWC) respectively, were synthetically analyzed. The results indicate that the formation and evolution of the modern warm current system in the ECS and the YS has been accompanied by the development of the KC and impulse rising of the sea level since the last deglaciation. The influence of the KC on the Okinawa Trough had enhanced since 16 cal kyr BP, and synchronously the modern TWC began to develop with the rising of sea level and finally formed at about 8.5 cal kyr BP. The KC had experienced two weakening process during the Heinrich event 1 and the Younger Drays event from 16 to 8.5 cal kyr BP. The period of 7-6 cal kyr BP was the strongest stage of the KC and the TWC since the last deglaciation. The YSWC has appeared at about 6.4 cal kyr BP. Thus, the warm current system of the ECS and the YS has ultimately formed. The weakness of the KC, indicated by the occurrence of Pulleniatina minimum event (PME) during the period from 5.3 to 2.8 cal kyr BP, caused the main stream of the TWC to shift eastward to the Pacific Ocean around about 3 cal kyr BP. The process resulted in the intruding of continent shelf cold water mass with rich nutrients. Synchronously, the strength of the YSWC was relatively weak and the related cold water body was active at the early-mid stage of its appearance against the PME background, which resulted in the quick formation of muddy deposit system in the southeastern YS. The strength of the warm current system in the ECS and the YS has enhanced evidently, and approached to the modern condition gradually since 3 cal kyr BP.

Sedimentation rates and provenance analysis in the Southwestern Okinawa Trough since the mid-Holocene

As a high-sedimentation rate depocenter along the path of the Kuroshio Current, the southwesternmost part of the Okinawa Trough is a key area to understand the Kuroshio history and sediments transportation. A 34.17-m-long sediment core was obtained by the advanced piston corer of Marco Polo/IMAGES XII MARION DUFRESNE during the May 2005 from the Southern Okinawa Trough at site MD05-2908. The recovered sediments were analyzed by AMS C-14 dating, coarse size fraction (> 63 mu m) extraction and moisture content determination in order to study its sedimentation flux and provenance. The depth-age relationship of core MD05-2908 was well constrained by 17 C-14 dating points. The sediments span across the mid-Holocene (6.8 ka B.P.) and have remarkablely high sedimentation rates between 1.8 and 21-2 m/ka, which is well consistent with the modern observations from sediment traps. We identified five 70-200 a periods of abnormally rapid sedimentation events at 6790-6600 a B.P., 5690-5600 a B.P., 4820-4720 a B.P., 1090-880 a B.P., and 260-190 a B.P., during which the highest sedimentation rate is up to 21-2 m/ka. In general, the lithology of the sediments were dominated by silt and clay, associated with less than 5% coarse size fraction (a parts per thousand << 63 mu m). As the most significant sediment source, the Lanyang River in northeastern Taiwan annually deliver about 10Mt materials to the coastal and offshore region of northeast Taiwan, a portion of which could be carried northward by currents toward the study area. Therefore, we concluded that the 5 abnormally rapid sedimentation events may be related to intensified rainfall in Taiwan and thus increased materials to our study area at that time. However, a few extreme-rapid sedimentation events cannot be explained by normal river runoff alone. The large earthquakes or typhoons induced hyperpycnal discharge of fluvial sediment to the ocean may also act as a potential source supply to the Okinawa Trough.

Relative paleointensity of the geomagnetic field during the past 200 ka from the West Philippine Sea and its chronological significance

Our rock magnetic analysis of core Ph05 from the West Philippine Sea demonstrates that the core preserves a strong, stable remanent magnetization and meets the magnetic mineral criteria for relative paleointensity (RPI) analyses. The magnetic minerals in the sequence are dominated by pseudosingle-domain magnetite, and the concentration of magnetic minerals is at the same scale. Both the conventional normalizing method and the pseudo-Thellier method were used in conjunction with the examination of the rock magnetic properties and natural remanent magnetization. Susceptibility (chi), anhysteretic remnant magnetization (ARM) and saturation isothermal remnant magnetization (SIRM) were used as the natural remanent magnetization normalizer. However, coherence analysis indicated that only ARM is more suitable for paleointensity reconstruction. The age model of core is established based on oxygen isotope data and AMS(14)C data, which is consistent with the age model estimated from RPI records. The relative paleointensity data provide a continuous record of the intensity variation during the last 200 ka, which correlates well with the global references RPI stacks. Several prominent low paleointensity values are identified and are correlated to the main RPI minima in the SINT-200 record, suggesting that the sediments have recorded the real changes of geomagnetic field.

Recent 2000-year geological records of mud in the inner shelf of the East China Sea and their climatic implications

AXIS(14)C dating and grain-size analysis for Core DD2, located at the north of the Yangtze River-derived mud off the Zhejiang-Fujian coasts in the inner shelf of the East China Sea, provide us a high-resolution grain-size distribution curve varying with depth and time. Data in the upper mud layer of Core DD2 indicate that there are at least 9 abrupt grain-size increasing in recent 2000 years, with each corresponding very well with the low-temperature events in Chinese history, which might result from the periodical strengthening of the East Asian Winter Monsoon (EAWM), including the first-revealed maximum temperature lowering event at around 990 a BP. At the same time, the finer grain size section in Core DD2 agrees well with the Sui-Tang Warming Period (600-1000 a AD) defined previously by Zhu Kezhen, during which the climate had a warm, cold and warm fluctuation, with a dominated cooling period of 750-850 a AD. The Little Ice Age (LIA) can also be identified in the core. It starts around 1450 a AD and was followed by a subsequent cooling events at 1510, 1670 and 1840 a AD. Timing of these cold events revealed here still needs to be further verified owing to some current uncertainty of dating we used in this study.

The history of the Yangtze River entering sea since the last glacial maximum: a review and look forward

This review paper provides a brief review on the development of ideas in the fields of the sea level change of the ECS (East China Sea), the history of the Yangtze River entering the sea and paleochannels in the shelf of the ECS since the Last Glacial Maximum (LGM). The paper summarizes two opposite theories about the Yangtze River entering the sea during the LGM. One theory is that the Yangtze River input a lacustrine in the north of Jiangsu province which was defunct in middle Holocene, and the river was once dry. The other was that the Yangtze River still existed and entered into the Okinawa Trough during the LGM, but scholars share different opinions on which course the river ran across and which place the river input the trough. This paper concludes future work is to study the evolution of the Yangtze River and the paleoclimate and the corresponding events as a whole from the view of regional and even global change, and more attention should be paid to the study on mud sediment, the Yangtze River's response to the changes in climate and sea-level, and the channel metamorphosis.