Seawater carbonate chemistry, sample density and Strongylocentrotus purpuratus size, filtering and respiration rate during experiments, 2011

Anthropogenic CO2 emissions are acidifying the world's oceans. A growing body of evidence is showing that ocean acidification impacts growth and developmental rates of marine invertebrates. Here we test the impact of elevated seawater pCO2 (129 Pa, 1271 µatm) on early development, larval metabolic and feeding rates in a marine model organism, the sea urchin Strongylocentrotus purpuratus. Growth and development was assessed by measuring total body length, body rod length, postoral rod length and posterolateral rod length. Comparing these parameters between treatments suggests that larvae suffer from a developmental delay (by ca. 8%) rather than from the previously postulated reductions in size at comparable developmental stages. Further, we found maximum increases in respiration rates of + 100 % under elevated pCO2, while body length corrected feeding rates did not differ between larvae from both treatments. Calculating scope for growth illustrates that larvae raised under high pCO2 spent an average of 39 to 45% of the available energy for somatic growth, while control larvae could allocate between 78 and 80% of the available energy into growth processes. Our results highlight the importance of defining a standard frame of reference when comparing a given parameter between treatments, as observed differences can be easily due to comparison of different larval ages with their specific set of biological characters.

Seawater carbonate chemistry and elemental composition of purple sea urchin (Strongylocentrotus purpuratus) calcite in a laboratory experiment

Ocean acidification will likely have negative impacts on invertebrates producing skeletons composed of calcium carbonate. Skeletal solubility is partly controlled by the incorporation of "foreign" ions (e.g. magnesium) into the crystal lattice of these skeletal structures, a process that is sensitive to a variety of biological and environmental factors. Here we explore effects of life stage, oceanographic region of origin, and changes in the partial pressure of carbon dioxide in seawater (pCO2) on trace elemental composition in the purple sea urchin (Strongylocentrotus purpuratus). We show that, similar to other urchin taxa, adult purple sea urchins have the ability to precipitate skeleton composed of a range of biominerals spanning low- to high-Mg calcites. Mg / Ca and Sr / Ca ratios were substantially lower in adult spines compared to adult tests. On the other hand, trace elemental composition was invariant among adults collected from four oceanographically distinct regions spanning a range of carbonate chemistry conditions (Oregon, Northern California, Central California, and Southern California). Skeletons of newly settled juvenile urchins that originated from adults from the four regions exhibited intermediate Mg / Ca and Sr / Ca between adult spine and test endmembers, indicating that skeleton precipitated during early life stages is more soluble than adult spines and less soluble than adult tests. Mean skeletal Mg / Ca or Sr / Ca of juvenile skeleton did not vary with source region when larvae were reared under present-day, global-average seawater carbonate conditions (400 µatm; pHT = 8.02 ± 0.03 1 SD; Omega calcite = 3.3 ± 0.2 1 SD). However, when reared under elevated pCO2 (900 µatm; pHT = 7.73 ± 0.03; Omega calcite = 1.8 ± 0.1), skeletal Sr / Ca in juveniles exhibited increased variance across the four regions. Although larvae from the northern populations (Oregon, Northern California, Central California) did not exhibit differences in Mg or Sr incorporation under elevated pCO2 (Sr / Ca = 2.10 ± 0.06 mmol/mol; Mg / Ca = 67.4 ± 3.9 mmol/mol), juveniles of Southern California origin partitioned ~8% more Sr into their skeletons when exposed to higher pCO2 (Sr / Ca = 2.26 ± 0.08 vs. 2.09 ± 0.005 mmol/mol 1 SD). Together these results suggest that the diversity of carbonate minerologies present across different skeletal structures and life stages in purple sea urchins does not translate into an equivalent geochemical plasticity of response associated with geographic variation or temporal shifts in seawater properties. Rather, composition of S. purpuratus skeleton precipitated during both early and adult life history stages appears relatively robust to spatial gradients and predicted future changes in carbonate chemistry. An exception to this trend may arise during early life stages, where certain populations of purple sea urchins may alter skeletal mineral precipitation rates and composition beyond a given pCO2 threshold. This potential for geochemical plasticity during early development in contrast to adult stage geochemical resilience adds to the growing body of evidence that ocean acidification can have differing effects across organismal life stages.

Seawater carbonate chemistry and Strongylocentrotus purpuratus body length and gene expression pattern changes during experiments, 2011

Extensive use of fossil fuels is leading to increasing CO2 concentrations in the atmosphere and causes changes in the carbonate chemistry of the oceans which represents a major sink for anthropogenic CO2. As a result, the oceans' surface pH is expected to decrease by ca. 0.4 units by the year 2100, a major change with potentially negative consequences for some marine species. Because of their carbonate skeleton, sea urchins and their larval stages are regarded as likely to be one of the more sensitive taxa. In order to investigate sensitivity of pre-feeding (2 days post-fertilization) and feeding (4 and 7 days post-fertilization) pluteus larvae, we raised Strongylocentrotus purpuratus embryos in control (pH 8.1 and pCO2 41 Pa e.g. 399 µatm) and CO2 acidified seawater with pH of 7.7 (pCO2 134 Pa e.g. 1318 µatm) and investigated growth, calcification and survival. At three time points (day 2, day 4 and day 7 post-fertilization), we measured the expression of 26 representative genes important for metabolism, calcification and ion regulation using RT-qPCR. After one week of development, we observed a significant difference in growth. Maximum differences in size were detected at day 4 (ca. 10 % reduction in body length). A comparison of gene expression patterns using PCA and ANOSIM clearly distinguished between the different age groups (Two way ANOSIM: Global R = 1) while acidification effects were less pronounced (Global R = 0.518). Significant differences in gene expression patterns (ANOSIM R = 0.938, SIMPER: 4.3% difference) were also detected at day 4 leading to the hypothesis that differences between CO2 treatments could reflect patterns of expression seen in control experiments of a younger larva and thus a developmental artifact rather than a direct CO2 effect. We found an up regulation of metabolic genes (between 10 to 20% in ATP-synthase, citrate synthase, pyruvate kinase and thiolase at day 4) and down regulation of calcification related genes (between 23 and 36% in msp130, SM30B, SM50 at day 4). Ion regulation was mainly impacted by up regulation of Na+/K+-ATPase at day 4 (15%) and down regulation of NHE3 at day 4 (45%). We conclude that in studies in which a stressor induces an alteration in the speed of development, it is crucial to employ experimental designs with a high time resolution in order to correct for developmental artifacts. This helps prevent misinterpretation of stressor effects on organism physiology.

I. Studies on alkaline phosphatase activity in developing sea urchin embryos. II. Studies on the activation of protein biosynthesis in sea urchin eggs at fertilization

I. Alkaline phosphatase activity in the developing sea urchin Lytechinus pictus has been investigated with respect to intensity at various stages, ionic requirements and intracellular localization. The activity per embryo remains the same in the unfertilized egg, fertilized egg and cleavage stages. At a time just prior to gastrulation (about 10 hours after fertilization) the activity per embryo begins to rise and increases after 300 times over the activity in the cleavage stages during the next 60 hours.

The optimum ionic strength for enzymatic activity shows a wide peak at 0.6 to 1.0. Calcium and magnesium show an additional optimum at a concentration in the range of 0.02 to 0.07 molar. EDTA at concentrations of 0.0001 molar and higher shows a definite inhibition of activity.

The intracellular localization of alkaline phosphatase in homogenates of 72-hour embryos has been studied employing the differential centrifugation method. The major portion of the total activity in these homogenates was found in mitochondrial and microsomal fractions with less than 5% in the nuclear fraction and less than 2% in the final supernatant. The activity could be released from all fractions by treatment with sodium deoxycholate.

II. The activation of protein biosynthesis at fertilization in eggs of the sea urchins Lytechinus pictus and Strongylocentrotus purpuratus has been studied in both intact eggs and cell-free homogenates. It is shown that homogenates from both unfertilized and fertilized eggs are dependent on potassium and magnesium ions for optimum amino acid incorporation activity and in the case of the latter the concentration range is quite narrow. Though the optimum magnesium concentrations appear to differ slightly in homogenates of unfertilized and fertilized eggs, in no case was it observed that unfertilized egg homogenates were stimulated to incorporate at a level comparable to that of the fertilized eggs.

An activation of amino acid incorporation into protein has also been shown to occur in parthenogenetically activated non-nucleate sea urchin egg fragments or homogenates thereof. This activation resembles that in the fertilized whole egg or fragment both in amount and pattern of activation. Furthermore, it is shown that polyribosomes form in these non-nucleate fragments upon artificial activation. These findings are discussed along with possible mechanisms for activation of the system at fertilization.

The Status of Eelgrass, Zostera marina, as Bay Scallop Habitat: Consequences for the Fishery in the Western Atlantic

Zostera marina is a member of a widely distributed genus of seagrasses, all commonly called eelgrass. The reported distribution of eelgrass along the east coast of the United States is from Maine to North Carolina. Eelgrass inhabits a variety of coastal habitats, due in part to its ability to tolerate a wide range of environmental parameters. Eelgrass meadows provide habitat, nurseries, and feeding grounds for a number of commercially and ecologically important species, including the bay scallop, Argopecten irradians. In the early 1930’s, a marine event, termed the “wasting disease,” was responsible for catastrophic declines in eelgrass beds of the coastal waters of North America and Europe, with the virtual elimination of Z. marina meadows in the Atlantic basin. Following eelgrass declines, disastrous losses were documented for bay scallop populations, evidence of the importance of eelgrass in supporting healthy scallop stocks. Today, increased turbidity arising from point and non-point source nutrient loading and sediment runoff are the primary threats to eelgrass along the Atlantic coast and, along with recruitment limitation, are likely reasons for the lack of recovery by eelgrass to pre-1930’s levels. Eelgrass is at a historical low for most of the western Atlantic with uncertain prospects for systematic improvement. However, of all the North American seagrasses, eelgrass has a growth rate and strategy that makes it especially conducive to restoration and several states maintain ongoing mapping, monitoring, and restoration programs to enhance and improve this critical resource. The lack of eelgrass recovery in some areas, coupled with increasing anthropogenic impacts to seagrasses over the last century and heavy fishing pressure on scallops which naturally have erratic annual quantities, all point to a fishery with profound challenges for survival.

History of Molluscan Fishery Regulations and the Shellfish Officer Service in Massachusetts

Oysters, Crassostrea virginica, and softshell clams, Mya arenaria, along the Massachusetts coast were harvested by European colonists beginning in the 1600’s. By the 1700’s, official Commonwealth rules were established to regulate their harvests. In the final quarter of the 1800’s, commercial fishermen began harvesting northern quahogs, Mercenaria mercenaria, and northern bay scallops, Argopecten irradians irradians, and regulations established by the Massachusetts Legislature were applied to their harvests also. Constables (also termed wardens), whose salaries were paid by the local towns, enforced the regulations, which centered on restricting harvests to certain seasons, preventing seed from being taken, and personal daily limits on harvests. In 1933, the Massachusetts Legislature turned over shellfisheries management to individual towns. Local constables (wardens) enforced the rules. In the 1970’s, the Massachusetts Shellfish Officers Association was formed, and was officially incorporated in 2000, to help the constables deal with increasing environmental problems in estuaries where fishermen harvest mollusks. The constables’ stewardship of the molluscan resources and the estuarine environments and promotion of the fisheries has become increasingly complex.

A Cooperative Management Solution to a Fishery Commons

A common property resource with open access, such as a fishery, will be used to excess when faced with sufficient demand. This will lead to an excessive amount of effort on the part of the fishery, resulting in a depletion of the stock. This paper discusses the development of a property rights regime for the Atlantic calico scallop, Argopecten gibbus, fishery of Florida. The management solution of the Calico Scallop Conservation Association (CSCA) provides an example of the assignment of property rights to a common property resource without resorting to governmental intervention. In this particular fishery, self-regulation limited early harvesting which would be uneconomic; there may be other fisheries in which self-regulation could be economically efficient and biologically appropriate. While this solution may not be applicable to all common property resources, for those cases which may be similar; the example of the CSCA provides valuable information that may be helpful in establishing a more efficient use of the resource. Some types of government facilitation may also be useful.

Contribución de las comunidades bentónicas en la dieta del róbalo (Eleginops maclovinus) en la Ría Deseado (Santa Cruz, Argentina)

Se analiza el aporte de la comunidad bentónica de la ría Deseado a la dieta del róbalo (Eleginops maclovinus), para contribuir al conocimiento de la trama alimentaria costera en la Patagonia austral. Entre la primavera 2005 y otoño 2006 se estudiaron las comunidades bentónicas submareales en áreas de pesca de E. maclovinus y paralelamente, se analizaron los contenidos alimentarios estomacales de róbalos provenientes de la pesca deportiva. La comunidad bentónica de planicies areno-fangosas fue dominada por poliquetos, representados principalmente por las familias Onuphidae, Orbiniidae y Maldanidae. Los crustáceos constituyeron el segundo grupo en importancia y estuvieron representados principalmente por los anfípodos gamáridos Heterophoxus sp. y Ampelisca sp. La comunidad submareal de fondos de rodados estuvo dominada por poliquetos de las familias Nereididae, Cirratulidae y Polynoidae, y los moluscos Perumytilus purpuratus y Margarites violacea. E. maclovinus presentó una dieta bentónica de tipo oportunista y generalista, con una tendencia hacia la ingesta de anfípodos gamáridos y algas clorofíceas. Durante la marea baja, la mayor contribución a su dieta la realizó la comunidad de planicies areno-fangosas submareales. Durante la marea alta, E. maclovinus se alimentó también en el intermareal rocoso, donde preda preferentemente las clorofíceas Enteromorpha spp. ENGLISH: The role of the benthic communities at Ría Deseado in the diet of the Patagonian blenny (Eleginops maclovinus) was analyzed in order to increase the understanding of the coastal food web in southern Patagonia. Subtidal benthic communities were surveyed between spring 2005 and autumn 2006 in areas of E. maclovinus sport fishing. Simultaneously, the stomach contents of patagonian blenny specimens caught during sport fishing were analyzed. The benthic community over flat sandy-muddy bottoms was dominated by polychaetes, mainly from the families Onuphidae, Orbiniidae and Maldanidae, followed by crustaceans, which were mainly represented by the gammarid amphipods Heterophoxus sp. and Ampelisca sp. The subtidal benthic community over pebbly bottoms was dominated by polychaetes from the families Nereididae, Cirratulidae and Polynoidae and the mollusks Perumytilus purpuratus and Margarites violacea. The diet of E. maclovinus was benthic opportunist and generalist, with a preference to feed on gammarid amphipods and chlorophycea algae. During low tide, the main dietary contribution came from the subtidal community over flat sandy-muddy bottoms whereas, during high tide, E. maclovinus also preyed on rocky intertidal species, mainly the Chlorophycea Enteromorpha spp.

First molluscan TNFR homologue in Zhikong scallop: Molecular characterization and expression analysis

Tumor necrosis factor receptors (TNFRs) are a superfamily of proteins characterized by the unique cysteine-rich domain (CRD) and their important roles in diverse physiological and pathological events such as inflammation, apoptosis, autoimmunity and organogenesis. The first member of the molluscan TNFR family, designated as CfTNFR, was identified from Zhikong scallop Chlamys farreri by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of CfTNFR was of 1334 bp, consisting of a 5' UTR of 17 bp, a 3'UTR of 69 by with a poly (A) tail, and an open reading frame (ORE) of 1248 by encoding a polypeptide of 415 amino acids with a theoretical isoelectric point of 8.33 and predicted molecular weight of 47.07 kDa. There were a signal peptide, a CRD, a transmembrane region and a death domain in the deduced amino acid sequence of CfTNFR, suggesting that it was a typical type 1 membrane protein. The high identities (22-40%) of CfTNFR with other TNFR superfamily members indicated that CfTNFR should be a member of TNFR superfamily, and moreover, it should be the first death domain-containing TNFR found in invertebrates. Phylogenetic analysis revealed that CfTNFR was closely related to TNFR-like proteins from Strongylocentrotus purpuratus, Drosophila melanogaster and Ciona intestinalis, and they formed a separate branch apart from vertebrate TNFRs. The spatial expression of CfTNFR transcripts in healthy and bacteria challenged scallops was examined by quantitative real-time PCR. CfTNFR transcripts could be detected in all tested tissues, including haemocytes, gonad, gill, mantle and hepatopancreas, and significantly up-regulated in the tissues of gonad, gill, mantle and hepatopancreas after Listonella anguillarum challenge, indicating that CfTNFR was constitutive and inducible acute-phase protein involved in immune defence. The present results suggested the existence of the TNFR-like molecules and TNF-TNFR system in low invertebrates, and provided new insights into the role of CfTNFR in scallop innate immune responses to invading microorganisms. (C) 2009 Elsevier Ltd. All rights reserved.

中国北方海域扇贝产品质量的比较研究

由于局部养殖环境质量恶化和病害频发的困扰，严重影响了我国扇贝养殖产量、质量和出口创汇。针对扇贝养殖面临产品质量不高等问题，系统评估我国扇贝产品质量十分必要。本文以我国北方海域海湾扇贝Argopecten irradians和栉孔扇贝Chlamys farreri为研究对象，较为系统地研究了大连湾、秦皇岛、莱州湾、烟台、威海、胶州湾、胶南等七海域扇贝质量，包括营养品质、重金属富集、麻痹性贝毒污染三个方面。 研究结果表明，栉孔扇贝的蛋白质营养优于海湾扇贝，而脂类营养则低于海湾扇贝，海湾扇贝在不同海域的主成分分析可以看出，地理位置接近的海域营养状况比较相似，各海域的综合营养品质依次为：莱州湾＞烟台＞大连湾＞秦皇岛＞威海＞胶州湾＞胶南； 重金属含量存在组织差异性，贝边的含量总是高于贝柱，取自同一海域的栉孔扇贝的重金属含量高于海湾扇贝，六种重金属的平均含量排序为：Zn＞Cu＞Cd＞As＞Pb＞Hg，部分海域扇贝Cd和Zn有超标现象，两种重金属分别超标1.41～17.28倍和1.24～1.59倍，Hg、Pb、As和Cu均未超标； 用小白鼠生物测试法对扇贝内脏团、贝柱和其他组织的贝毒含量进行测定，结果仅在秦皇岛和大连湾海湾扇贝的内脏团中检测出麻痹性贝毒，其毒力分别为2.17 Mu•g -1和2.48 Mu•g -1，未超过食用安全标准（4 Mu•g -1），其他海域的样品均未检测到麻痹性贝毒的存在。

栉孔扇贝与海湾扇贝免疫学比较研究

扇贝是我国重要的海水养殖品种，但自1997年以来，养殖扇贝陆续爆发的大规模死亡，不但造成了巨大的经济损失，而且直接威胁到现有产业的生存和发展。引起扇贝大规模死亡原因是多方面的，其主要原因是病原滋生、养殖环境恶化和种质衰退。因此，深入研究扇贝免疫防御机制，探讨提高机体抗病力的有效途径和方法，改良种质和培育抗病品系，促进我国扇贝养殖业的健康、可持续发展。本文通过比较栉孔扇贝和海湾扇贝正常状态和重金属污染胁迫下相关免疫指标的变化及HSP70和HSP90在重金属胁迫后表达规律，以期更好的了解贝类的防御机制，为扇贝病害防治提供资料。 本研究采用流式细胞仪技术对栉孔扇贝和海湾扇贝血细胞死亡率、细胞吞噬率和呼吸爆发进行了测定，发现正常状态下两种扇贝的细胞死亡率相差不大，呼吸爆发的基础值相差也不大，而海湾扇贝细胞吞噬率显著高于栉孔扇贝。在血淋巴和肝胰腺中，海湾扇贝SOD、ACP酶活性均显著高于栉孔扇贝；海湾扇贝的MDA含量也高于栉孔扇贝，但差异不显著。鳗弧菌感染实验发现，海湾扇贝累积死亡率远低于栉孔扇贝。上述结果表明，海湾扇贝对细菌等异物的吞噬和杀灭能力以及机体自身的抗氧化能力高于栉孔扇贝，这为海湾扇贝比栉孔扇贝具有更高的抗逆性提供了证据。 不同浓度Pb2+刺激后，海湾扇贝血细胞的死亡率明显低于栉孔扇贝。海湾扇贝细胞吞噬率和呼吸爆发均高于栉孔扇贝。对两种扇贝体液免疫指标的测定发现，海湾扇贝的SOD、ACP活性和MDA含量均高于栉孔扇贝。结果表明，在Pb2+刺激下两种扇贝都处于重金属氧化胁迫下，其免疫系统均受到了影响，但相同剂量的Pb2+对两种扇贝的毒害程度不同，海湾扇贝受到的影响要低于栉孔扇贝。 采用同源克隆和cDNA 末端快速扩增（RACE）技术，从海湾扇贝血淋巴中克隆到了热休克蛋白90（AiHSP90）基因。AiHSP90的cDNA序列全长为2669bp，其中5' 非编码区（UTR）为90bp，3' UTR为44bp，开放阅读框（Open Reading Frame, ORF）包括2175bp，2524-2528位AATAA为推测的AiHSP90基因的 mRNA 多聚腺苷酸加尾信号（Polyadenylation Signal）。开放阅读框编码724个氨基酸残基，推测的理论分子量为83.08 kDa，等电点为4.81。 利用荧光实时定量PCR 技术检测了AiHSP90基因在革兰氏阴性菌鳗弧菌和革兰氏阳性菌藤黄微球菌感染后不同时间点相对于β-actin基因的表达。在鳗弧菌和藤黄微球菌刺激后，除72h外AiHSP90均上调表达，在9h其mRNA表达达到最大值，分别约为空白组的6.8倍和3倍；随后随着时间的推移，其表达下降，在48h恢复到接近空白组的水平；到72h时降到最低点。鳗弧菌对AiHSP90的诱导比藤黄微球菌要高，这也反映出鳗弧菌对海湾扇贝有更强的毒力，诱导的HSP90表达也更强。本研究首次揭示出在软体动物中HSP90在细菌感染条件下的表达情况，结果表明海湾扇贝AiHSP90参与了机体对细菌感染的应答。 采用荧光实时定量PCR技术，对栉孔扇贝和海湾扇贝在不同浓度、不同种类重金属胁迫后HSP70和HSP90基因表达规律进行了比较。结果显示，镉和铅对栉孔扇贝处理10天后即诱导HSP70以较高水平表达，20天后镉处理组HSP70恢复到与对照接近的水平，而在铅处理组也呈现出下降的趋势。而海湾扇贝HSP70基因表达与栉孔扇贝恰好相反，处理20天后其HSP70表达才出现较高水平。铜离子处理对诱导两种扇贝HSP70的影响相似，对栉孔扇贝HSP70的诱导强度更大。对于同一种重金属，不同的浓度诱导的HSP70表达也各不相同。而对于HSP90，海湾扇贝在受到三种重金属刺激后HSP90的表达量均高于栉孔扇贝，而且长时间的刺激（20天）诱导了更高的HSP90基因表达。不同的浓度的同一种重金属离子诱导的HSP90表达也各不相同。这些结果表明，在重金属胁迫下HSP70以及HSP90在栉孔扇贝和海湾扇贝中均表现出不同的应答方式。结合前面海湾扇贝抗逆性较强的综合结果，两种扇贝热休克蛋白基因表达的比较提示我们栉孔扇贝HSP70的激烈反应预示着抗逆能力的不足，而海湾扇贝HSP90的高表达却预示着更强的抗逆能力，有待于更多应激实验如热刺激、病原刺激等的进一步验证。

海湾扇贝微卫星标记的开发及遗传连锁图谱的构建

海湾扇贝（Argopecten irradians）不连续分布于美国大西洋沿岸和墨西哥湾沿岸，自1982年以来北部亚种（A. i. irradians）和南部亚种（A. i. concentricus）被先后引进到中国，由于其生长速度快、繁殖周期短和适宜温度范围广的特点，迅速成为我国海水养殖的重要品种。近年来飞速发展的分子标记技术为优良品种的选育注入了新的活力，相对于传统的表型选择来说，标记辅助选择不易受环境的影响，尤其是对于低遗传力性状和后期表达的性状，能增强选择效率，提高选择的准确度，缩短育种周期。本文通过构建海湾扇贝微卫星富集文库获得大量的微卫星DNA序列，筛选多态的微卫星标记构建了海湾扇贝的遗传连锁图谱，并应用复合区间作图法对生长相关性状进行了QTL定位。 本研究利用富集文库-菌落原位杂交法筛选海湾扇贝微卫星DNA，吸附(AC)15和(AG)15探针的尼龙膜捕捉并富集含有微卫星序列的片段，菌落原位杂交结果显示阳性克隆率达到40%，测序比对后获得521个独立的阳性克隆，其中微卫星506个，小卫星15个。微卫星中，完美型248个，占49.0%，非完美型216个，占42.7%；复合型42个，占8.3%；AG/TC重复占大多数（356个，70.4%），AC/TG重复有150个（29.6%）。设计合成了382对引物，利用38个海湾扇贝个体对其中15个微卫星位点进行了遗传多样性评价，不同位点扩增得到的等位基因数从3到7个不等，期望杂合度和观测杂合度的范围分别为0.198~0.813和0.083~0.833，实验结果表明富集文库-菌落原位杂交法适合大规模筛选微卫星标记。 利用8个微卫星标记对海湾扇贝1个野生种群和3个养殖群体的遗传多样性与分化进行了比较和分析。8个位点共扩增得到35个等位基因，平均每个位点4.38个等位基因，平均有效等位基因数为2.30，平均观测杂合度和期望杂合度分别为0.41和0.46。相比于野生群体（美国），养殖群体（北卡罗来那、浙江和胶南）的等位基因数和杂合度都有所降低，在封闭环境下养殖19代的浙江群体等位基因数丢失最严重，共有9个等位基因丢失（25.7%）。经过多代人工养殖后，海湾扇贝养殖与野生群体之间和养殖群体之间出现了明显的遗传分化，胶南群体与野生群体的遗传距离最大，而胶南群体与浙江群体的遗传距离已经超过了胶南群体（北部亚种）和北卡罗来那（南部亚种）群体的遗传距离，这种分化将有利于海湾扇贝的杂交选育。 利用167个微卫星标记和1个壳色标记，以海湾扇贝2个全同胞F1代为作图群体，构建了海湾扇贝的性别遗传连锁图谱。整合的雌性连锁图谱含有118个标记，覆盖了16个连锁群，每个连锁群含有的标记数目从4到16个不等，平均每个连锁群上有7.4个标记，图谱总长度为761.0 cM，标记间的平均间隔为8.55 cM，图谱的覆盖率为73.5%；整合雄性连锁图谱含有126个标记，覆盖了17个连锁群，每个连锁群含有的标记数目从2到11个不等，平均每个连锁群上有7.4个标记，图谱总长度为729.1 cM，标记间的平均间隔为6.75 cM，图谱的覆盖率为74.7%。雌性亲本的重组率高于雄性，雌雄亲本共享标记间的重组率比值为1.18：1。偏分离标记在性别间呈现不对称分布，雄性亲本的偏分离高于雌性亲本，可能与雄性亲本来源于亚种间杂交的遗传背景相关。 利用海湾扇贝微卫星遗传连锁图谱在两个作图家系中对5个生长性状的QTL进行了定位，5个生长性状的表型相关均达到极显著水平（P < 0.01），Pearson相关系数均超过0.781，总重、壳长、壳宽、壳高和壳重的QTL（LOD > 2.0）的数目分别为8、6、6、7和6个。这些QTL成簇分布于CC5家系的LG1、LG3、LG4、LG8和CC10家系的LG1、LG3、LG6、LG8、LG9连锁群，单个QTL可解释的表型方差为5.5%到29.2%，QTL成簇分布现象说明这些生长相关的性状可能具有共同的遗传基础，家系特异性QTL暗示在不同的遗传背景和环境下存在不同的主效QTL。本研究定位的QTL，尤其在两个家系中共享的QTL为下一步分子标记辅助选择提供了参考区间。