近江牡蛎3个种群生化遗传变异的研究

用同工酶电泳方法研究广东省官渡、阳江和汕尾的近江牡蛎（Ostea rivularis）的遗传变异。分析了11种同工酶20个的位点，其中出现13个位点显示为多态。结果表明官渡、阳江、汕尾种群的多态位点比例分别为0．200、0．200、0．500。官渡和阳江种群间，官渡和汕尾种群间以及阳江和汕尾种群间的遗传距离（D）分别为0．0451，0．1125,0．1088。     

近江牡蛎的染色体组型

以近江牡蛎的担轮幼虫为材料,采用秋水仙素——低渗处理——空气干燥制片法制作染色体标本,对其核型进行分析,结果表明,近江牡蛎体细胞染色体数目2n=20,NF=40,核型公式为20m。     

近江牡蛎3个种群生化遗传变异的研究

用同工酶电泳方法研究广东省官渡、阳江和汕尾的近江牡蛎(Ostea rivularis)的遗传变异。分析了11种同工酶20个的位点,其中出现13个位点显示为多态。结果表明官渡、阳江、汕尾种群的多态位点比例分别为0.200、0.200、0.500。官渡和阳江种群间,官渡和汕尾种群间以及阳江和汕尾种群间的遗传距离(D)分别为0.0451,0.1125,0.1088。     

近江牡蛎肠道细菌及其产酶能力

从近江牡蛎肠道中分离出21株菌,其中, 9株为革兰阳性菌, 12株革兰阴性菌。为寻找能分泌多种酶的有益微生物,研究了它们蛋白酶、脂肪酶、淀粉酶、纤维素酶的产酶能力。结果表明,有61 .9%的菌株能分泌蛋白酶或淀粉酶(各有13株菌);有52. 4%的菌株能分泌脂肪酶(11株菌);有33. 3%的菌株能产纤维素酶(7株菌)。在这21株菌中,产4种酶的有5株菌,产3种酶的3株菌,产两种酶的5株菌,产1种酶的3株菌,不产酶的仅有5株菌。不产酶的菌株仅占细菌总数的23. 8%,而各种产酶菌则高达76. 2%,由此表明细菌在近江牡蛎对食饵消化过程的重要性。     

近江牡蛎肠道细菌及其产酶能力

从近江牡蛎肠道中分离出21株菌，其中，9株为革兰阳性菌，12株革兰阴性菌。为寻找能分泌多种酶的有益微生物，研究了它们蛋白酶、脂肪酶、淀粉酶、纤维素酶的产酶能力。结果表明，有61．9％的菌株能分泌蛋白酶或淀粉酶(各有13株菌)；有52．4％的菌株能分泌脂肪酶(11株菌)；有33．3％的菌株能产纤维素酶(7株菌)。在这21株菌中，产4种酶的有5株菌，产3种酶的3株菌，产两种酶的5株菌，产1种酶的3株菌，不产酶的仅有5株菌。不产酶的菌株仅占细菌总数的23．8％，而各种产酶菌则高达76．2％，由此表明细菌在近江牡蛎对食饵消化过程的重要性。     

不同方式提取的近江牡蛎糖蛋白的体外抗氧化活性

比较研究了水浸提、盐液浸提及超声辅助盐液浸提的近江牡蛎糖蛋白粗提物对羟自由基、DPPH自由基、超氧阴离子自由基的清除效果。结果表明:水浸提、盐液浸提及超声辅助盐液浸提的糖蛋白中可溶性蛋白与总糖含量之比分别为1.83∶1、4.49∶1和2.51∶1;3种糖蛋白粗提物均具有一定的清除羟自由基、DPPH自由基和超氧阴离子自由基的能力;它们对羟自由基的清除能力高于对DPPH自由基和超氧阴离子自由基;盐液浸提物清除超氧阴离子的能力最弱;超声辅助盐液浸提的糖蛋白粗提物清除羟自由基、超氧阴离子自由基及DPPH自由基的能力比水浸提和盐液浸提的糖蛋白的能力强。     

湛江港区底泥悬浮物对近江牡蛎的毒害效应

模拟湛江港涉海工程施工产生的悬浮物,探讨底泥悬浮物对工程附近海区近江牡蛎养殖的影响,研究不同浓度悬浮物对大、小规格近江牡蛎(Crassostrea rivularis)的毒害效应。结果表明,悬浮物对大规格近江牡蛎(8.12±1.76 cm)的毒害影响不明显;对小规格近江牡蛎(2.33±0.78 cm),悬浮物浓度≤270 mg/L时死亡率低于30%,悬浮物浓度≥810 mg/L时死亡明显增加,死亡率达60%。通过概率单位法计算,小规格近江牡蛎的96 h半致死浓度为627.17 mg/L。     

近江牡蛎两个野生种群的遗传多样性分析

采用RAPD分子标记和rDNA-ITS1序列分析了近江牡蛎Crassostrea rivularis湛江官渡和阳江程村野生种群的遗传多样性。12个RAPD引物共扩增出8838条片段,157个位点,平均每个引物可产生13个位点,片段长度在200～2200bp之间。湛江种群和阳江种群的多态位点比例分别为89.62%和89.57%,遗传多样性指数分别为0.4170和0.4334。种群间平均遗传距离为0.0327,平均遗传相似性为0.9681,平均遗传分化系数为0.0437。得到近江牡蛎18S、5.8S部分序列和ITS1全部序列,其中ITS1序列片段长度为478～485bp,共有11个变异位点,两个为转换(A/G),其他为插入/缺失(A/-、T/-)。湛江和阳江种群各获得8个单倍型,其中有一个单倍型为两个种群共享。两个种群的CG碱基平均含量较高,分别为58.29%和58.41%。种群间的遗传分化系数0.0254。结果说明,近江牡蛎湛江种群和阳江种群间具有较高的遗传多样性和较低的遗传分化。     

近江牡蛎耗氧率和排氨率的研究

采用室内实验生态学方法研究近江牡蛎的耗氧率和排氨率。结果表明,在实验温度13~33℃范围内,近江牡蛎的耗氧率(RO)和排氨率(RN)与体重(W)均呈负相关,可用Y=aW-b表示。耗氧率和温度的关系可表示为RO=-c+b1t-b2 t2,耗氧率为0.626~2.354 mg.g-1.h-1;在温度13~28℃范围内,耗氧率随温度的升高而增加,28℃时,耗氧率达最大值,温度升高至33℃时,耗氧率反而下降。排氨率与温度的关系可表示为RN=c1e d1t,排氨率为0.075~0.318 mg.g-1.h-1,且随温度的升高,排氨率呈持续升高趋势。近江牡蛎呼吸和排泄Q10值范围分别为0.631~3.399和1.046~2.288。在28℃,不同规格近江牡蛎呼吸氧原子数与排出氨氮原子数的比值(nO/nN)均达到最大值。方差分析表明,体重、温度及二者的交互作用对近江牡蛎的耗氧率和排氨率均有极显著的影响(P<0.01)。近江牡蛎的日常代谢明显高于标准代谢,耗氧率和排氨率平均值分别提高46.8%和67.7%。     

Pigment Distribution and Secretion in the Mantle of the Pacific Oyster(Crassostrea gigas)

The color of Mollusca shells is one of the most important attributes to consumers. At the cellular level, black color is mainly from the melanin produced by melanocytes. The melanosome is a specialized membrane-bound organelle that is involved in melanin synthesis, storage, and transportation. How the complex pigmentation process in the Crassostrea gigas is established remains an open question. The objectives of this studies are to examine the morphological characteristics of melanosomes or mela...     

Relationship Between Shell Color and Growth and Survival Traits in the Pacific Oyster Crassostrea gigas

Shell color is regarded as an economic trait in many breeding programs of bivalve mollusks, but the relationship between shell color and production traits remains controversial. In our breeding program of Crassostrea gigas, solid white, black, and orange shell lines were obtained, and second-generation(F2) and testcross families with segregating shell colors were constructed by crossing these three lines. These segregating families provided ideal samples for investigating the relationship between shell color and production traits in C. gigas. The growth and survival of 7-month-old oysters with different shell colors sampled within the same families were compared in seven F2 families and 13 testcrosses. In addition, the growth and survival of oysters from the three shell color lines were compared at 4 and 16 months of age. The growth and survival rates of the orange shell line were significantly lower than those of the white and black shell lines. However, no significant difference in growth between oysters with different shell colors was observed within segregating families, except the testcrosses produced by crossing orange-and white-shelled grandparents, and no significant difference in survival was observed in any family. Overall, no significant correlation was observed between shell color and production traits in C. gigas. These results suggest that shell color cannot be used as a marker to guide the selection of growth and survival traits. Thus shell color and production traits should be selected independently in oyster breeding programs.     

Linkage Disequilibrium in Wild and Cultured Populations of Pacific Oyster(Crassostrea gigas)

Linkage disequilibrium(LD) can be applied for mapping the actual genes responsible for variation of economically important traits through association mapping.The feasibility and efficacy of association studies are strongly dependent on the extent of LD which determines the number and density of markers in the studied population,as well as the experimental design for an association analysis.In this study,we first characterized the extent of LD in a wild population and a cultured mass-selected line of Pacific oyster(Crassostrea gigas).A total of 88 wild and 96 cultured individuals were selected to assess the level of genome-wide LD with 53 microsatellites,respectively.For syntenic marker pairs,no significant association was observed in the wild population;however,three significant associations occurred in the cultured population,and the significant LD extended up to 12.7 c M,indicating that strong artificial selection is a key force for substantial increase of genome-wide LD in cultured population.The difference of LD between wild and cultured populations showed that association studies in Pacific oyster can be achieved with reasonable marker densities at a relatively low cost by choosing an association mapping population.Furthermore,the frequent occurrence of LD between non-syntenic loci and rare alleles encourages the joint application of linkage analysis and LD mapping when mapping genes in oyster.The information on the linkage disequilibrium in the cultured population is useful for future association mapping in oyster.     

粤西镇海湾近江牡蛎线粒体16S rRNA基因序列变异分析

采用聚合酶链式反应 (PCR)技术对粤西镇海湾水域的近江牡蛎Crassostrearivularis(Gould)群体 2 7个个体的线粒体DNA16SrRNA基因序列片段进行扩增 ,获得大约 5 0 0bp的扩增产物。PCR产物经纯化后进行序列测定 ,经ClustalX同源排序 ,除去引物及部分端部序列 ,得到4 14bp的核苷酸片段。 2 7个个体共检测到 2个变异位点 ,均为颠换位点 ,没发现碱基位点插入、缺失及转换位点 ,共 3种单倍型 ,每个单倍型只有一个碱基的差异。运用DNASP软件计算得该群体的核苷酸多样性和平均核苷酸差异数分别为 0 .0 0 0 36和 0 .14 815。此结果提示镇海湾近江牡蛎群体遗传多样性已很低 ,很有必要从其他分布区引进近江牡蛎亲贝来扩大该种群的遗传多样性     

Effects of Salinity and Temperature on Growth and Survival of Juvenile Iwagaki Oyster Crassostrea nippona

Iwagaki oyster Crassostrea nippona occurs naturally along the coasts of Japan and Korea. Because of its unique flavor, delicious taste, edibility during the summer and high commercial value, it has been identified as a potential aquaculture species. To determine the optimum aquaculture conditions and provide necessary information for mass production of the juvenile, the effects of six salinities(15, 20, 25, 30, 35 and 40) and five temperatures(16, 20, 24, 28 and 32℃) on growth and survival of juvenile C. nippona were examined in this study. In the salinity experiment, the largest values of mean shell height and growth rate were observed at salinity 25(20.96 ± 0.36 mm and 172.0 μm d~(-1), respectively), which were significantly different(P 0.05) with those of other treatments, except at salinity 30(20.56 ± 1.05 mm and 160.3 μm d~(-1), respectively)(P 0.05). The maximum survival rate 84.44% was always observed at salinity 20, and there was no significant difference(P 0.05) in survival rate among salinities varying between 15 and 35. In the temperature-related experiments, the highest growth and survival rates of juvenile were observed at 24℃(180.8 μm d~(-1) and 84.4%) and 28℃(190.7 μm d~(-1) and 83.3%), respectively, on day 20, and showed significantly(P 0.05) larger size and higher survival rate than any other groups. Both juvenile survival and growth were significantly depressed at extreme salinities(15, 40) and temperatures(16℃, 32℃). Based on the results of the present study, a salinity range from 25 to 30 and a temperature range from 24℃ to 28℃ are considered optimal conditions for survival and growth of juvenile C. nippona.     

Mass Selection for Growth Improvement in Black Shell Line of Pacific Oyster Crassostrea gigas

The Pacific oyster, Crassostrea gigas, naturally distributing along the coast of northwest Pacific, is one of the most important bivalve species due to its high economic value and fecundity. In China, we have initiated a selective breeding program on both shell color and growth rate of C. gigas since 2010. A black shell line was obtained through four-generation family selection. In this study, mass selection for growth improvement was conducted in the sixth generation and seventh generation of black shell lines. To assess the progress of potential genetic improvement, the progeny of two generations of black shell lines were selected to evaluate their shell heights via a 450-day farming experiment. As the results, after growing for 450 days, the sixth generation and seventh generation of selected lines were 9.03% and 11.42% larger than the control lines, respectively. During the grow-out stage, the genetic gain of two generations was 8.82% ± 0.18% and 11.54% ± 0.43%, respectively; and the corresponding realized heritability was 0.45 ± 0.04 and 0.41 ± 0.04, respectively. These results indicated that the mass selection for shell height achieved steady progress in the two generations of C. gigas.     

Biochemical Composition and Nutritional Value of Different Shell Color Strains of Pacific Oyster Crassostrea gigas

The shell color of Pacific oyster(Crassostrea gigas) is a desirable trait; but the nutritional studies on C. gigas with different shell colors have not been conducted. Through successive selective breeding, five shell color strains of black(B), purple(P), orange(O), golden(G) and white(W) C. gigas have been developed. The aim of this study was to evaluate the chemical composition and nutritional value of five shell color strains and one commercial population with a common color. The biochemical composition including moisture, total protein, glycogen, ash, total fat, fatty acids(FA), amino acids and minerals was detected. The results indicated that the protein(50.76%–56.57%) was the major component. The content of glycogen showed a significant difference between orange shell and golden shell strains, as well as between commercial population and golden shell strain. In addition, all shell color strains contained a large amount of essential amino acids(12.20–14.15 g(100 g)~(-1)), of them leucine(2.81–3.29 g(100 g)~(-1)) and lysine(2.79–3.28 g(100 g)~(-1)) were predominant. The oysters were rich in polyunsaturated fatty acids(42.26%–45.24% of total fatty acid) with high levels of DHA(18.53%–21.16% of total fatty acid) and EPA(17.23%–18.68% of total fatty acid). Significant differences of mineral contents(Mg, Zn, Fe and Cu) were identified among the six populations. These results indicated that C. gigas with different shell colors presented rich nutritional value with high protein, glycogen, essential amino acids and polyunsaturated fatty acids. The biochemical composition obtained in this study is useful for selective breeding of C. gigas with different shell colors.