大口黑鲈选育群体遗传结构的微卫星分析

采用微卫星标记技术分析不同世代大口黑鲈选育群体的遗传结构。利用11对微卫星引物对大口黑鲈第2～4代选育群体及南水群体(对照)共120个个体进行PCR扩增,共得到28个等位基因。每个位点获得2～3个等位基因,平均等位基因为2.54。第2代(F_2)、第3代(F_3)、第4代(F_4)及南水群体(CG)的平均多态信息含量(PIC)值分别为:0.423、0.419、0.386、0.366。与F_2相比,F_4的遗传多样性减少8.76%,但与CG相比,F_4仍具有较高的遗传多样性,表明大口黑鲈选育群体的遗传基础逐步趋向纯化,且仍有较大选育潜力。此外,尽管F_2与F_4的遗传距离(0.022)比F_2与F_3的(0.011)大,但世代间的F_(st)值(0.006～0.009)均小于0.05,且世代群体总遗传分化指数为0.013,表明选育群体已出现遗传分化,但分化程度较低。     

环境胁迫对栉孔扇贝(Chlamys farreri)血浆蛋白及其抗菌活性的影响

采集采自山东某养殖场的栉孔扇贝(Chlamys farreri)健康贝和病贝的血淋巴,测定其血浆蛋白浓度、血浆蛋白图谱和相对分子质量,以及血浆抗菌活性、凝集素活性等几种重要的体液免疫因素,研究环境胁迫对血浆蛋白及其抗菌活性的影响。结果表明:栉孔扇贝(C.farreri)血浆蛋白质量浓度约1 260~1 650μg/mL;血浆蛋白SDS-PAGE图谱显示3条蛋白带,相对分子质量分别为34 000、25 000和14 000;健康贝血浆抗菌活性指数BI为0.34,病贝BI为0.21;凝集素活力水平为1.602~2.204。氨氮胁迫下血浆蛋白浓度下降约780μg/mL,凝集素活力下降为1.301;高温胁迫下血浆蛋白质量浓度下降约为900μg/mL,凝集素活力为1.903;高盐胁迫下血浆蛋白浓度下降约为990μg/mL,凝集素活力为2.204。     

Isozyme expression of Chinese and Japanese populations of Chlamys farreri and their reciprocal hybrids

Chinese and Japanese population of Chlamys farreri and their reciprocal hybrids were surveyed in isozyme variability at 13 loci by polyacrytamide gel electrophoresis (PAGE). Isozyme banding patterns indicated these hybrids were diploid. Loci that were observed as being monomorphic in inbred populations of C. farreri were also found to be monomorphic in filial progeny; loci that observed to be polymorphic in parental type generations were also polymorphic in hybrid generations. Differences existed among allelic frequency of the four types of cross. Within the reciprocal hybrids the expression of malic enzyme (ME) isozyme was sufficient to distinguishing individual hybrids because of the band, Rf=0.38. However, there were no noticeable variations among all the samples to differentiate one from another. Inbreeding was likely to be the main problem in aquaculture. The introduction of new broodstock can improve the genetic diversity. Hybrid vigor has manifested to a certain extent in the present study.     

Immunohistochemical identification of dynorphin A and Kappa opioid receptor-1 in the digestive system of scallop Chlamys farreri

Journal of Oceanology and Limnology - Little is known about the roles of dynorphin and Kappa opioid receptor (KOR) in mollusks. In this study, we aim to determine the distribution of dynorphin A...     

Biomarker responses in the bivalve Chlamys farreri to the water-soluble fraction of crude oil

To investigate the effect of the water soluble fraction of crude oil(WSF) on marine bivalves, the scallop C hlamys farreri was exposed to three WSF concentrations(0.18 mg/L, 0.32 mg/L, and 0.51 mg/L, respectively) in seawater. Petroleum hydrocarbon contents in scallops and a suite of enzymes [7-Ethoxyresorufin-O-deethylase(EROD), aryl hydrocarbon hydroxylase(AHH), glutathione S-transferase(GST), and glutathione peroxidase(GPx)] in gills and digestive glands were monitored over 10 days. The results revealed that WSF affected the activity of the four enzymes in the gills and digestive glands. EROD activity in the gills was significantly induced in most individuals of the three test groups, while in the digestive gland it was significantly induced in the low-concentration group within 4 days but was inhibited in the middle- and high-concentration groups on days 1, 4, and 10. AHH activity in the gills of all treatment groups was significantly induced on day 1. In the digestive gland, AHH activity was induced in most individuals from the treatment groups. In all treatment groups, GST activity was significantly inhibited from days 2 to 10 in the gills and was induced after day 4 in the digestive gland. GPx activity in the gills was significantly inhibited throughout the exposure period in all treatment groups. There was no overall significant difference in GPx activity in the digestive gland between the control and treatment groups. Our results also revealed that petroleum hydrocarbon concentrations in the tissues increased linearly with exposure time. EROD activity in the digestive gland and GST and GPx activity in the gill tissue were negatively correlated with petroleum hydrocarbon body burden. These enzymes play important roles in detoxification and can act as potential biomarkers for monitoring petroleum hydrocarbon contaminants in the marine environment.     

Mapping Toll-Like Receptor Signaling Pathway Genes of Zhikong Scallop (Chlamys farreri) with FISH

Toll-like receptor(TLR) signaling pathway plays a pivotal role in the innate immune system. Studies on TLR signaling pathway genes in Zhikong scallop(Chlamys farreri) have mainly focused on sequence analysis and expression profiling, no research has been carried out on their localization. The chromosomal position of TLR signaling pathway genes can be valuable for assemblying scallop genome and analysizing gene regulatory networks. In the present study, five key TLR signaling pathway genes(Cf TLR, Cf Myd88, Cf TRAF6, Cf NFκB, and Cf IκB) containing bacterial artificial chromosomes(BACs) were isolated and physically mapped through fluorescence in situ hybridization on five non-homologous chromosome pairs, showing a similar distribution to another five model species. The isolation and mapping of these key immune genes of C. farreri will aid to the research on innate immunity, assignment of interested genes to chromosomes, and integration of physical, linkage and cytogenetic maps of this species.     

Effects of polypeptide from Chlamys farreri on amino acid con-tent in guinea pig skin irradiated by chronic ultraviolet A and B

1 INTRODUCTION Long-term exposure of the skin to ultraviolet (UV) radiation results in degenerative processes involved in photoaging and photocarcinogenesis (Differy, 1987). As UVC is absorbed entirely by the stratospheric ozone layer and does not reach the earth抯 surface, sunlight is composed of UVA (320 nm 400 nm) and UVB (290 nm 320 nm). Human skin is exposed constantly to both UVA and UVB radiation, it is, therefore, important to protect the skin against UVA and UVB-induce…     

Development of 101 novel EST-derived single nucleotide polymorphism markers for Zhikong scallop (Chlamys farreri)

Zhikong scallop(Chlamys farreri) is an important maricultured species in China.Many researches on this species,such as population genetics and QTL fine-mapping,need a large number of molecular markers.In this study,based on the expressed sequence tags(EST),a total of 300 putative single nucleotide polymorphisms(SNPs) were selected and validated using high resolution melting(HRM) technology with unlabeled probe.Of them,101(33.7%) were found to be polymorphic in 48 individuals from 4 populations.Further evaluation with 48 individuals from Qingdao population showed that all the polymorphic loci had two alleles with the minor allele frequency ranged from 0.046 to 0.500.The observed and expected heterozygosities ranged from 0.000 to 0.925 and from 0.089 to 0.505,respectively.Fifteen loci deviated significantly from Hardy-Weinberg equilibrium and significant linkage disequilibrate was detected in one pair of markers.BLASTx gave significant hits for 72 of the 101 polymorphic SNP-containing ESTs.Thirty four polymorphic SNP loci were predicted to be non-synonymous substitutions as they caused either the change of codons(33 SNPs) or pretermination of translation(1 SNP).The markers developed can be used for the population studies and genetic improvement on Zhikong scallop.     

Importance of kelp-derived organic carbon to the scallop Chlamys farreri in an integrated multi-trophic aquaculture system

Bivalves and seaweeds are important cleaners that are widely used in integrated multi-trophic aquaculture(IMTA) systems. A beneficial relationship between seaweed and bivalve in the seaweedbased IMTA system has been confirmed, but the trophic importance of seaweed-derived particulate organic materials to the co-cultured bivalve is still unclear. We evaluated the trophic importance of the kelp Saccharina japonica to the co-cultured scallop Chlamys farreri in a typical IMTA farm in Sungo Bay(Weihai, North China). The dynamics of detritus carbon in the water were monitored during the culturing period. The proportion of kelp-derived organic matter in the diet of the co-cultured scallop was assessed via the stable carbon isotope method. Results showed that the detritus carbon in the water ranged from 75.52 to 265.19 μg/L, which was 25.6% to 73.8% of total particulate organic carbon(TPOC) during the study period. The amount of detritus carbon and its proportion in the TPOC changed throughout the culture cycle of the kelp. Stable carbon isotope analysis showed that the cultured scallop obtained 14.1% to 42.8% of its tissue carbon from the kelp, and that the percentages were closely correlated with the proportion of detritus carbon in the water( F =0.993, P= 0.003). Evaluation showed that for 17 000 tons(wet weight) of annual scallop production, the kelp contributed about 139.3 tons of carbon(535.8 tons of dry mass). This confirms that cultured kelp plays a similar trophic role in IMTA systems as it does in a natural kelp bed. It is a major contributor to the detritus pool and supplies a vital food source to filter-feeding scallops in the IMTA system, especially during winter and early spring when phytoplankton are scarce.     

Identification of Cytochrome P450 (CYP) genes in Zhikong scallop (Chlamys farreri)

Cytochrome P450 (CYP) superfamily is one of the membership largest and function most diverse protein superfamily recogniozed among living beings. Members of this superfamily were further assigned to different families and subfamilies based on their amino acid similarities. According to their phylogenetic relationships, the CYP genes which likely diverged from common ancestor gene and may share common functions were grouped into one clan. Widely distributing scallops are a group of the most conspicuous bivalve; however the studies on their CYP is acarce. In this study, we searched the genome and expressed sequence tags of Zhikong scallop (Chlamys farreri) for CYP genes. In total, 88 non-redundant CYP were identified, which were homed in 13 CYPs gene families. Phylogenetic analysis divided these genes into 4 CYP clans. As in deuterostomes, Clan 2 was the largest, which contained 33 genes belonging to CYP1, CYP2, CYP17 and CYP356 families. Clan 3 contgained 19 genes belonging to CYP3, CYP5 and CYP30 families. Clan 4 contained 23 genes, all belonging to CYP4 family. The mitochondrial CYP clan contained 9 genes belonging to CYP10 and CYP24 families. In comparison, protostomes (C. farreri, D. pluex, D. melanogaster) contained more CYP genes than deuterostomes (S. purpuratus and vertebrates) in Clan 2 but less genes in Clan 3 and Clan 4. Our findings will aid to deciphering CYP function and evolution in scallops and bivalves.     

Effects of benzo（a）pyrene exposure on the antioxidant enzyme activity of scallop Chlamysfarreri

Scallop Chlamys farreri was exposed to different concentrations of benzo(a)pyrene (BaP) (0.5 μg/L, 1.0 μg/L, 10.0 μg/L and 50.0 μg/L) for 30 days in seawater. The 7-ethoxyresorufin O-deethylase (EROD) activity was significantly induced, and increased with the increasing BaP concentration. The glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), Glutathione peroxidase (GPx) activities increased in short time at low concentration of BaP, and was significantly depressed at high concentr...     

军曹鱼全人工繁殖群体遗传特征的SSR分析

利用8个微卫星DNA位点分析南海海域5个军曹鱼全人工繁育群体(HN1、HN2、ZJ、FJ和LS)子代的遗传多样性特征和群体间遗传分化。结果显示,军曹鱼养殖群体与天然群体的遗传结构特征基本一致:1)平均有效等位基因数为3.910±0.440,观测杂合度为0.595±0.049,分子方差分析(AMOVA)表明,个体内分化占主导(46%),军曹鱼养殖群体整体遗传多样性较高;2)群体间基因流明显(N_m=2.5959,F_(st)=0.0878),整体分化程度较低。各养殖群体表现出不同于天然大群体的特征:1)绝大部分位点均明显偏离哈温平衡,杂合子缺失或过剩现象普遍存在;2)聚类和群体分配分析等表明HN2与另四个养殖群体(HN1、ZJ、FJ和LS)分化明显。     

Polypeptide from Chlamys farreri inhibits UVB-induced apoptosis of HaCaT cells via iNOS/NO and HSP90

Polypeptide from Chlamys farreri(PCF) is a novel marine bioactive product that was isolated from the gonochoric Chinese scallop Chlamys farreri,and was found to be an effective antioxidant in our recent studies.In this study,we investigated the effect of PCF on ultraviolet B(UVB)-induced apoptosis of HaCaT cells and the intracellular signaling pathways involved.Pretreatment with the inducible nitric oxide synthase(iNOS) inhibitor S-methylisothiourea sulfate inhibited UVB-induced apoptosis,indicating that iN...     

Protective Effect of Topically Applied Polypeptide from Chlamys farreri Against Ultraviolet Radiation-Induced Chronic Skin Damage in Guinea Pig

INTRODUCTIONSolarultraviolet(UV)radiationisknowntobeamajoretiologicalriskofphotoaging.AsUVC(2 0 0nm -2 90nm)isabsorbedentirelybythestratosphericozonelayer,sunlightontheearthsur faceiscomposedofUVA (3 2 0nm -4 0 0nm)andUVB (2 90nm -3 2 0nm)only.UVAradiationenergyaccountsformorethan 90 %ofsolarUV ;andpenetratesthebasallayerofepidermismoreefficientlythanUVB (Brulsetal.,1984) .However,atsimilardosages,UVBismuchmoremuta genicthanUVA (Drobetskyetal.,1995) .Furthermore ,humanskinisexp…