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Xu D.,Marine Fishery Institute of Zhejiang Province | Xu D.,Zhejiang Ocean University | Lou B.,Marine Fishery Institute of Zhejiang Province | Bertollo L.A.C.,Federal University of São Carlos | Cioffi M.D.B.,Federal University of São Carlos
Molecular Cytogenetics | Year: 2013

Despite the theoretical and experimental progress, our understanding on sex chromosome differentiation is still diagrammatic. The accumulation of repetitive DNA sequences is believed to occur in early stages of such differentiation. As fish species present a wide range of sex chromosome systems they are excellent models to examine the differentiation of these chromosomes. In the present study, the chromosomal distribution of 9 mono-, di- and tri-nucleotide microsatellites were analyzed using fluorescence in situ hybrization (FISH) in rock bream fish (Oplegnathus fasciatus), which is characterized by an X1X2Y sex chromosome system. Generally, the males and females exhibited the same autosomal pattern of distribution for a specific microsatellite probe. The male specific Y chromosome displays a specific amount of distinct microsatellites repeats along both arms. However, the accumulation of these repetitive sequences was not accompanied by a huge heterochromatinization process. The present data provide new insights into the chromosomal constitution of the multiple sex chromosomes and allow further investigations on the true role of the microsatellite repeats in the differentiation process of this sex system. © 2013 Xu et al.; licensee BioMed Central Ltd.


Xu D.,Marine Fishery Institute of Zhejiang Province | Xu D.,Zhejiang Ocean University | Lou B.,Marine Fishery Institute of Zhejiang Province | Xu H.,Marine Fishery Institute of Zhejiang Province | And 2 more authors.
Marine Biotechnology | Year: 2013

Sex-specific DNA markers applicable were very useful for elucidating the sex-determination mechanism and sex control in fishes. In the present study, amplified fragment-length polymorphism (AFLP) approach with 144 primer combinations was employed to identify sex-specific markers in the rock bream. Four male-specific AFLP fragments were identified which were designated as Opl286, Opl237, Opl422, and Opl228. Further sequence analysis of the sex markers' genomic region revealed subtle differences between the males and females. We identified four male-specific single-nucleotide polymorphisms (SNPs) and a deletion of 8 bp in marker Opl286, six male-specific SNPs in marker Opl237, three male-specific SNPs in marker Opl422, and eight male-specific SNPs and 1 bp inversions in marker Opl228. Specific primers were designed based on the nucleotide variation in the sequences to develop a simple polymerase chain reaction method for identifying the genetic sex of rock bream. As a result, three out of the four male-specific markers were converted into SNP markers. The male-specific AFLP markers and AFLP-derived SNP markers were tested in 100 individuals collected from three locations around the coast of Zhoushan, yielding reproducible sex identification. These male-specific DNA markers are a useful tool for the identification of the sex-determining locus in rock bream and for guiding artificial breeding programs. © 2012 Springer Science+Business Media, LLC.


PubMed | National Taiwan Ocean University, Marine Fishery Institute of Zhejiang Province, Zhejiang Ocean University, Chung Yuan Christian University and CAS Qingdao Institute of Oceanology
Type: | Journal: Genomics data | Year: 2016

The rock bream (Oplegnathus fasciatus) is considerably one of the most economically important marine fish in East Asia and has a unique neo-Y chromosome system that is a good model to study the sex determination and differentiation in fish. In the present study, we used Illumina sequencing technology (HiSeq2000) to sequence, assemble and annotate the transcriptome of the testis and ovary tissues of rock bream. A total of 40,004,378 (NCBI SRA database SRX1406649) and 53,108,992 (NCBI SRA database SRX1406648) high quality reads were obtained from testis and ovary RNA sequencing, respectively, and 60,421 contigs (with average length of 1301bp) were obtained after de novo assembling with Trinity software. Digital gene expression analysis reveals 14,036 contigs that show gender-enriched expressional profile with either testis-enriched (237 contigs) or ovary-enriched (581 contigs) with RPKM >100. There are 237 male- and 582 female-abundant expressed genes that show sex dimorphic expression. We hope that the gonad transcriptome and those gender-enriched transcripts of rock bream can provide some insight into the understanding of genome-wide transcriptome profile of teleost gonad tissue and give useful information in fish gonad development.


Hu H.,Marine Fishery Institute of Zhejiang Province | Sun X.,Marine Fishery Institute of Zhejiang Province | Zhong Z.,Marine Fishery Institute of Zhejiang Province | Chen X.,Marine Fishery Institute of Zhejiang Province | And 2 more authors.
Journal of Separation Science | Year: 2012

A simple, efficient, solvent-free, and commercial readily available approach for determination of 11 chlorobenzenes (CBs) in water samples using the static headspace (HS) sampling and gas chromatography-electron capture detector has been described. The proposed static HS sampling method was initially optimized and the optimum experimental conditions found were 10 mL water sample containing 20% (w/v) sodium chloride placed in a 20 mL vial and stirred at 70°C for 30 min. The linearity of the method ranged from 0.16 to 8.0 μg/L for dichlorobenzene isomers, 0.0176∼0.88 μg/L for trichlorobenzene isomers, 0.004∼0.2 μg/L for tetrachlorobenzene isomers, and from 0.001 to 0.05 μg/L for pentachlorobenzene and hexachlorobenzene, with correlation coefficients ranging between 0.9992 and 0.9999. The limits of detection were in the low μg/L level, ranging between 0.0002 and 0.04 μg/L. The relative recoveries of spiked CBs with external calibration or standard addition method at different concentration levels in pure, tap, and sea water samples were 83∼116%, 89∼108%, and 93∼112%, respectively, and with relative standard deviations of 1.9∼6.3%, 1.6∼5.4%, and 2.5∼5.7% (n = 5), respectively. It is concluded that this method can be successfully applied for the determination of CBs in pure, tap, and sea water samples. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Zhang X.,University of North Carolina at Chapel Hill | Hu H.,Marine Fishery Institute of Zhejiang Province
Central European Journal of Chemistry | Year: 2014

In addition to its role as a carrier of genetic information, DNA has been recognized as a construction material for the assembly of different objects and structural arrangements with nanoscale features. As a result of DNA's self-recognition properties (based on the specific base-pairing of G-C and T-A), monolayer films of nucleic acids on solid supports have attracted an escalating attentions. Recently, numerous novel materials based on two-dimensional (2D) and three-dimensional (3D) DNA structures have been reported, which extends their utility to a large number of appliations. This review paper intends to be a new and comprehensive overview of recent strategies to site- specifically immobilized DNA on various materials, including carbonaceous substances, gold, and silica substrate, emphasizing the applications of site-specific DNA nanostructure-based devices for diagnostic, bioanalytical, food safety and environmental monitoring. Additionally, an up-to-date perspective is proposed at the end of this review. © Versita Sp. z o.o.


Xu D.,Marine Fishery Institute of Zhejiang Province | Lou B.,Marine Fishery Institute of Zhejiang Province | Shi H.,Marine Fishery Institute of Zhejiang Province | Geng Z.,Marine Fishery Institute of Zhejiang Province | And 2 more authors.
Biochemical Systematics and Ecology | Year: 2012

The genetic diversity and population genetic structure of Nibea albiflora were investigated using mitochondrial cytochrome c oxidase subunit I (COI) gene sequences. A total of 108 individuals from four localities around the China Sea were included in the analysis. Overall, 21 polymorphic sites were observed and 19 haplotypes were defined. The N. albiflora populations were characterized by medium/high haplotype diversity (0.580-0.815) and low nucleotide diversity (0.00125-0.00219). Analysis of neutral evolution and mismatch distribution implied that N. albiflora might have experienced a recent population expansion. Pairwise fixation index (Fst) analysis indicated significant genetic differentiation among populations from different localities. The hierarchical analysis of molecular variance (AMOVA) analysis also showed significant genetic divergence ( st = 0.0233, P < 0.01) among these populations. The present results suggest that N. albiflora populations around the China Sea have developed significantly divergent genetic structures. This study provides new information for genetic assessment and will be crucial for establishing fisheries management and strategies for this species. © 2012 Elsevier Ltd.


Zhang X.-N.,Paine College | Hu H.-M.,Marine Fishery Institute of Zhejiang Province
Wuli Huaxue Xuebao/ Acta Physico - Chimica Sinica | Year: 2016

The alignment of the ionic liquid (IL) cation and anion at the interface is of interest because it would affect the surface structures and properties of IL at the interfaces. In this study, Kelvin probe force microscopy (KPFM), a scanning probe microscopy technique, was used to investigate the interfacial properties of the IL at room temperature. A model molecule, 1-butyl-3-methylimidazolium chloride ([Bmim]Cl), was selectively assembled on the lyophilic chemical patterns prepared on a substrate, forming ultrathin solid-like adsorbate layers and droplets. Because the surface potential is a direct indicator of the surface dipole, which is useful for examining molecular orientation, the surface potential maps captured by KPFM indicated that the [Bmim]Cl molecules demonstrated different orientations at the gas-liquid interface (in the form of a droplet) and at the gassolid interface (in the form of a solid-like adsorbate layer). Our results indicate that KPFM has potential for the characterization of IL molecular alignment at interfaces. © Editorial office of Acta Physico-Chimica Sinica.


Hu H.,Marine Fishery Institute of Zhejiang Province | Li Z.,Marine Fishery Institute of Zhejiang Province | Zhang X.,Marshall University | Xu C.,Hanshan Normal University | Guo Y.,Marine Fishery Institute of Zhejiang Province
Journal of Separation Science | Year: 2013

A method was developed for the rapid separation of catecholamines by nonaqueous microchip electrophoresis (NAMCE) with LIF detection, A homemade pump-free negative pressure sampling device was used for rapid bias-free sampling in NAMCE, the injection time was 0.5 s and the electrophoresis separation conditions were optimized. Under the optimized conditions, the samples were separated completely in <1 min. The average migration times of the epinephrine (E), dopamine (DA), and norepinephrine (NE) were 34.26, 43.81, and 50.07 s, with an RSD of 1.05, 1.26, and 0.89% (n = 7), respectively. The linearity of the method ranged from 0.0125 to 2.0 mg/L for E and 0.025∼4.0 mg/L for DA and NE, with correlation coefficients ranging between 0.9978 and 0.9986. The detection limits of E, DA, and NE were 2.5, 5.0, and 5.0 μg/L, respectively. The recoveries of E, DA, and NE in spiked urine samples were between 86 and 103%, with RSDs of 4.5∼6.8% (n = 5). The proposed NAMCE with LIF detection combined with a pump-free negative pressure sampling device is a simple, inexpensive, energy efficient, miniaturized system that can be successfully applied for the determination of catecholamines in urine samples. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Hu H.-M.,Zhejiang University | Hu H.-M.,Marine Fishery Institute of Zhejiang Province | Yin X.-F.,Zhejiang University | Wang X.-Z.,Zhejiang University | And 2 more authors.
Journal of Separation Science | Year: 2013

This article describes a further development of our previously reported miniaturized analysis system of microchip electrophoresis with on-line chemiluminescence detection. The system, developed first time for nonaqueous microchip electrophoresis with peroxyoxalate chemiluminescence detection, consists of a suction pressure device for sample or reagent introduction, a constant voltage supplied for electrophoretic separation, an either hydrophilic or hydrophobic porous polymer plug for preventing chemiluminescence reagent flowing upstream and a spiral detection channel for enhancement of both detection sensitivity and reproducibility. Especially, by using organic solvent as BGE medium, the developed system avoided the interface problem between aqueous running buffer and low-water-content chemiluminescence solvent in previous reports. The influencing factors on chemiluminescence signal were optimized using rhodamine 6G as model molecule. The system performance was further investigated in the experiment of separation of hydrophilic rhodamine dyes and analysis of hydrophobic polycyclic aromatic hydrocarbon, providing the detection limit (S/N = 3) of 3.5 nmol/L for rhodamine 123, 6.8 nmol/L for rhodamine 6G, and 60 nmol/L for 1-aminopyrene, respectively. The experimental results showed the system offered a number of benefits, including compact structure, high sensitivity, good reproducibility, and a wide range of application prospect. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


PubMed | Paine College and Marine Fishery Institute of Zhejiang Province
Type: Journal Article | Journal: Journal of separation science | Year: 2016

A simple, efficient, solvent-free, and commercial readily available approach for determination of five volatile chlorinated hydrocarbons in water samples using the static headspace sampling and gas chromatography with electron capture detection has been described. The proposed static headspace sampling method was initially optimized and the optimum experimental conditions found were 10 mL water sample containing 20% w/v sodium chloride placed in a 20 mL vial and stirred at 50C for 20 min. The linearity of the method was in the range of 1.2-240 g/L for dichloromethane, 0.2-40 g/L for trichloromethane, 0.005-1 g/L for perchloromethane, 0.025-5 g/L for trichloroethylene, and 0.01-2 g/L for perchloroethylene, with coefficients of determination ranging between 0.9979 and 0.9990. The limits of detection were in the low g/L level, ranging between 0.001 and 0.3 g/L. The relative recoveries of spiked five volatile chlorinated hydrocarbons with external calibration method at different concentration levels in pure, tap, sea water of Jiaojiang Estuary, and sea water of waters of Xiaomendao were in the range of 91-116, 96-105, 86-112, and 80-111%, respectively, and with relative standard deviations of 1.9-3.6, 2.3-3.5, 1.5-2.7, and 2.3-3.7% (n = 5), respectively. The performance of the proposed method was compared with traditional liquid-liquid extraction on the real water samples (i.e., pure, tap, and sea water, etc.) and comparable efficiencies were obtained. It is concluded that this method can be successfully applied for the determination of volatile chlorinated hydrocarbons in different water samples.

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