Changes of Biogenic Elements in Phragmites australis and Suaeda salsa from Salt Marshes in Yellow River Delta,China

Little information is available on biogenic elements(carbon, nitrogen, phosphorus and sulfur) and the ecological stoichiometric characteristics of plants in coastal wetlands. To investigate the contents of carbon, nitrogen, phosphorus and sulfur of plants, and their ecological stoichiometric characteristics in the Yellow(Huanghe) River Delta, plant samples were collected from two typical salt marshes(Suaeda salsa and Phragmites australis wetlands) during the period of from August to October in 2007, and the ratios of C/N, C/P, N/P, C/N/P and C/N/P/S were calculated. Results showed that during the studying period, plant C, N and P were lower than the global average values, and plant N and P were lower than the China's average values. Leaf C and S in Suaeda salsa were significantly lower than those in Phragmites australis(P 0.05), and leaf N and P in Suaeda salsa and Phragmites australis showed no significant differences(P 0.05). Average C/N ratios were 23.75 in leaf, 73.36 in stem, 65.67 in root of Suaeda salsa, and 33.77 in leaf, 121.68 in stem, 97.13 in root of Phragmites australis. Average C/N ratios of Suaeda salsa and Phragmites australis were all great than 25, indicating the salt marsh in the Yellow River Delta is an N limitation system. Average C/P ratios were 276.78 in leaf, 709.28 in stem and 1031.32 in root of Suaeda salsa, and 536.94 in leaf, 768.13 in stem and 875.22 in root of Phragmites australis. The average N/P ratios of Suaeda salsa were 12.92 in leaf, 10.77 in stem and 10.91 in root, and the average N/P ratios of Phragmites australis were 16.40 in leaf, 7.40 in stem and 6.92 in root, indicating the Suaeda salsa wetlands were N limited and Phragmites australis wetlands were N limited in August and P limited in October in 2007. The average C/N, C/P and C/N/P ratios in Suaeda salsa and Pragmites australis were higher than the global average values, indicating the lower quality of organic matter provided by wetland plants in the Yellow River delta.     

Diurnal expression of circadian clock genes period 1 and period 3 in Pelteobagrus vachellii

Circadian clock genes are crucial for generating and sustaining most rhythmic daily functions in the animal kingdom,which entrain the rhythms of biochemical,physiological,and behavioural processes.To better understand the molecular oscillations of the circadian rhythms in darkbarbel catfish(Pelteobagrus vachellii),we isolated and characterized two circadian clock genes in P.vachellii,period 1(perl),and period3(per3).The circadian clock gene perl was found to encode a 1 428-amino acid polypeptide,including PER-ARNT-SIM(PAS) dimerisation domains,a PAS-associated C-terminal motif(PAC),a short mutable domain(S/M),and a nuclear export signal(NES).The 4 902-bp per3 cDNA includes an open reading frame encoding a 1 292-amino acid residue polypeptide with a PER-ARNT-SIM(PAS) domain,cytoplasmic localisation domain(CLD),interaction site(TIS),and a nuclear localisation signal(NLS).The perl and per3 gene was constitutively expressed in all examined tissues.Moreover,perl expression within a light/dark cycles showed rhythmic expression in the diencephalon,brain,liver and intestine,with,the acrophase at 15:15,12:52,7:51,and 12:55,respectively.Daily expression of per3 was rhythmic in the diencephalon,brain,liver and intestine,with the acrophase at 8:15,9:54,10:39,and 10:25 h,respectively.These findings expand our understanding of circadian mechanism at the molecular level in this species.     

Molecular cloning and mRNA expression analysis of myosin heavy chain (MyHC) from fast skeletal muscle of grass carp, Ctenopharyngodon idella

The myosin heavy chain (MyHC) is one of the major structural and contracting proteins of muscle. We have isolated the cDNA clone encoding MyHC of the grass carp, Ctenopharyngodon idella. The sequence comprises 5 934 bp, including a 5 814 bp open reading frame encoding an amino acid sequence of 1 937 residues. The deduced amino acid sequence showed 69% homology to rabbit fast skeletal MyHC and 73%–76% homology to the MyHCs from the mandarin fish, walleye pollack, white croaker, chum salmon, and carp. The putative sequences of subfragment-1 and the light meromyosin region showed 61.4%–80% homology to the corresponding regions of other fish MyHCs. The tissue-specific and developmental stage-specific expressions of the MyHC gene were analyzed by quantitative real-time PCR. The MyHC gene showed the highest expression in the muscles compared with the kidney, spleen and intestine. Developmentally, there was a gradual increase in MyHC mRNA expression from the neural formation stage to the tail bud stage. The highest expression was detected in hatching larva. Our work on the MyHC gene from the grass carp has provided useful information for fish molecular biology and fish genomics.     

Heavy metals in sediments of Yellow Sea and East China Sea: Chemical speciation,distribution, infl uence factor,and contamination

Metal species and the degree of environmental pollution are related to the hydrogen sulfi de(H 2 S), an important product of early diagenesis that can react with metals to form stable compounds. To investigate the eff ects of H 2 S to metals and evaluate metal environment eff ect in the sediments of the East China Sea(ECS) and Yellow Sea(YS), geochemical characteristic and spatial distribution of nine heavy metals and H 2 S profi le were studied. Higher H 2 S content and lower metal content was observed in the sediments 10 cm in depth in the North Yellow Sea and the west coast of South Korea. The pollution load index( I pl) indicates that the southern coast of Shandong Peninsula underwent moderate pollution( I pl =1) of heavy metals and no heavy metal pollution appeared in other areas( I pl 1). To some extents, the ecological risk of Cd and As enrichment was moderately severe in all stations. The chance of heavy metal combination to be toxic in ECS and YS during summer was 21%. In addition, correlation between H 2 S content and metals in both solid and porewater phases was obvious, corroborating important eff ect of H 2 S on metal distribution. Moreover, H 2 S could aff ect the spatial distributions of heavy metals in porewater directly and be indicative of potential biological eff ects of combined toxicant groups in the study region.     

Molecular Characterization and Expression and DNA Methylation Analyses of a Galectin-Related Protein Gene from Cynoglossus semilaevis

Galectins,a family of ?-galactoside-binding proteins,participate in both innate immunity and adaptive immunity.This study identified one novel galectin-related protein from half-smooth tongue sole Cynoglossus semilaevis,which was designated as Cs GRP.The full-length c DNA of its encoding gene was 785 bp in length with a 528 bp open reading frame encoding a putative protein of 176 amino acids.The deduced Cs GRP contained a putative 131-aa galactoside-binding lectin domain and 3 critical residues responsible for carbohydrate binding(R~(93),W~(109) and R~(114)).Genomic structural analysis revealed that Cs GRP consisted of five exons and four introns.Cs GRP showed 68% similarity with Poecilia latipinna GRP and 67% similarity with Stegastes partitus GRP.Cs GRP showed the highest expression level in liver,although its expression was detected in all tested tissues.When challenged with Vibrio harveyi,the expression of Cs GRP was significantly down-regulated in liver(P 0.05).In addition,we found that in spleen and kidney of C.semilaevis,the Cp G island of Cs GRP showed significantly higher(P 0.05) methylation level in disease-resistant family of C.semilaevis(DR-Cs) than in disease-susceptible ones(DS-Cs).Our results suggested that Cs GRP may play important roles in the immune response of C.semilaevis.Moreover,DNA methylation patterns provided valuable data for understanding the relationship between epigenetic regulation and immunity,which would assist the animal genetic research and improve the animal breeding in the future.     

口蹄疫病毒VP1植物表达载体的构建

P1 T重组质粒上含有口蹄疫病毒 (FMDV)GD10分离株的p1cDNA片段 ,以此为模板 ,用PCR方法扩增其中的VP1基因 ,获得大小约 6 40bp的片段。该片段用BglⅡ和BstEⅡ酶切消化后克隆至表达载体 pCAMBIA130 5 .2 ,转化EcoliTOP10感受态细胞。重组质粒经PCR、酶切及序列分析 ,证实VP1基因处于CaMV35S启动子控制 ,且读码框正确     

Construction of cDNA subtractive library from pearl oyster (Pinctada fucata Gould) with red color shell by SSH

The molecular basis of color polymorphism in the shells of the pearl oyster Pinctada fucata is largely unknown. We developed a red-shelled family line and used suppression subtractive hybridization (SSH) to screen for differentially expressed genes in red- and non-red-shelled pearl oysters. We constructed forward and reverse cDNA subtractive libraries consisting of 2 506 and 797 clones, respectively. Among 343 randomly selected clones in the forward library, 304 sequences were identified in GenBank using BLASTx and BLASTn. Of the 304 sequences, 13 showed no similarity to known sequences and 291 were matched with known genes of the pearl oyster, including shematrin-1, shematrin-2, shematrin-6, shematrin-7, nacrein, nacrein-like protein, aspein for shell matrix protein, glycine-rich protein, mantle gene 5, 28S, EST00031, EST00036, 16S, and COΙ. In the reverse library, 7 clones were sequenced and analyzed by BLAST. Two sequences shared similarity with EST00036 from the P. fucata subtraction cDNA library, four with the P. fucata mitochondrial gene for 16S rRNA and 1 with P. fucata shematrin-2. We evaluated the expression of 12 genes from the forward library using RT PCR. Two sequences matched with 16S and COΙ so were considered to be false positives. The remaining 10 sequences were differentially expression in the red-shelled pearl oysters. Our results suggest that differential expression of these genes may be related to color variation in the red-shelled family line of the pearl oyster.     

Potential Effects of Episodic Deposition on Nutrients and Heavy Metals in Decomposing Litters of Suaeda glauca in Salt Marsh of the Yellow River Estuary,China

Episodic deposition has been recognized as a major factor affecting the decomposition rate of detrital material in salt marshes. In this paper, three one-off burial treatments, no burial treatment(0 cm, NBT), current burial treatment(10 cm, CBT) and strong burial treatment(20 cm, SBT), were designed in intertidal zone of the Yellow River Estuary to determine the potential influences of episodic deposition on nutrient(C, N) and heavy metal(Pb, Cr, Cu, Zn, Ni, Mn, Cd, V and Co) variations in decomposing litters of Suaeda glauca. Results showed that although various burial treatments showed no statistical difference in decomposition rate of S. glauca, the values generally followed the sequence of CBT(0.002 403/d) SBT(0.002 195/d) NBT(0.002 060/d). The nutrients and heavy metals in decomposing litters of the three burial treatments exhibited different variations except for N, Cu, Cr, Ni and Co. Except for Mn, no significant differences in C, N, Pb, Cr, Cu, Zn, Ni, V and Co concentrations occurred among the three treatments(P 0.05). With increasing burial depth, Cr and Cd levels generally increased while Cu, Ni and Mn concentrations decreased. Although episodic deposition was generally favorable for C and N release from S. glauca, its influence on release was insignificant. In the three burial treatments, Pb, Cr, Zn, Ni, Mn, V and Co stocks in S. glauca generally evidenced the export of metals from litter to environment, and, with increasing burial depth, the export amounts increased greatly. The S. glauca were particular efficient in binding Cd and releasing Pb, Cr, Zn, Ni, Mn, V and Co, and, with increasing burial depth, stocks of Cu in decomposing litters generally shifted from release to accumulation. The experiment indicated that the potential eco-toxic risk of Pb, Cr, Zn, Ni, Mn, V and Co exposure would be serious as the strong burial episodes occurred in S. glauca marsh.     

Molecular Cloning and Expression ofPoIR2, a Novel Gene Involved in Immune Response in Japanese Flounder （Paralichthys olivaceus）

A novel immune-related gene was expressed in Japanese flounder (Paralichthys olivaceus) injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5’UTR, a 336 bp open reading frame (ORF) encoding 111 amino acids and a 556bp 3’UTR. Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S (NMS), but further alignment indicated they did not have NMS C-terminal conservational signature domain. So it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa signal peptide and was a secretory pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after infected with V. anguillarum. These results indicated that the PoIR2 might play some important role in Japanese flounder immune response system. This gene was named PoIR2 (P.olivaceus immune-related gene 2, GenBank accession number: EU224372). The mature PoIR2 peptide was expressed in BL21(DE3) pLysS using pET-32a(+) vector and a great part of the recombinant mature peptide existed as soluble type.     

Biosorption of Cr (VI), Cr (III), Pb (II) and Cd (II) from aqueous solutions by Sargassum wightii and Caulerpa racemosa algal biomass

Heavy metal pollution is one of the most important environmental problems today.Biosorption is an innovative tech-nology that employs biological materials to accumulate heavy metals from waste water through metabolic process or physicochemi-cal pathways of uptake.Even though several physical and chemical methods are available for removal of heavy metals,currently many biological materials such as bacteria,algae,yeasts and fungi have been widely used due to their good performance,low cost and large quantity of availability.The aim of the present study is to explore the biosorption of toxic heavy metals,Cr(VI),Cr(III),Pb(II) and Cd(II) by algal biomass obtained from algae Sargassum wightii(brown) and Caulerpa racemosa(green).Biosorption of algal biomass was found to be biomass concentration-and pH-dependent,while the maximal biosorption was found at pH 5.0 and with the metal concentration of 100 mg L-1.S.wightii showed the maximal metal biosorption at the biomass concentration of 25 g L-1,followed by C.racemosa with the maximal biosorption at 30 g L-1.S.wightii showed 78% biosorption of Cr(VI),Cr(III),Pb(II) and Cd(II) ions.C.racemosa exhibited 85% biosorption of Cd(II) and Cr(VI),and 50% biosorption of Cr(III) and Pb(II).The results of our study suggest that seaweed biomass can be used efficiently for