长牡蛎dpp同源基因的克隆及其在贝壳发生中的功能研究

dpp是转化生长因子β家族的重要成员,在胚胎发育中作为形态发生子参与体轴形成及附肢发育等过程。为了研究牡蛎dpp同源基因在贝壳发生中的作用,揭示贝壳发生的分子机理,作者克隆了一种长牡蛎(Crassostrea gigas)的dpp基因,命名为cgdpp。序列和进化分析显示,cgdpp分子中不同位点的氨基酸残基变异程度不同。作者用整装原位杂交的手段研究了dpp同源基因在长牡蛎早期发育中的表达情况。结果表明,dpp同源基因参与了从贝壳形成区开始分化到早期贝壳形成的全过程。在担轮幼虫(Trochophore)中,dpp同源基因似乎调控了贝壳的形状与扩张速度;在早期D形幼虫中,dpp同源基因表达量突然下降至痕量水平,并与贝壳发育区无明显的关联,提示dpp同源基因可能仅仅参与了贝壳的发生,不参与其进一步的发育过程。     

Genome-wide survey and analysis of microsatellites in the Pacific oyster genome: abundance,distribution, and potential for marker development

Microsatellites are a ubiquitous component of the eukaryote genome and constitute one of the most popular sources of molecular markers for genetic studies. However, no data are currently available regarding microsatellites across the entire genome in oysters, despite their importance to the aquaculture industry. We present the fi rst genome-wide investigation of microsatellites in the Pacifi c oyster Crassostrea gigas by analysis of the complete genome, resequencing, and expression data. The Pacifi c oyster genome is rich in microsatellites. A total of 604 653 repeats were identifi ed, in average of one locus per 815 base pairs(bp). A total of 12 836 genes had coding repeats, and 7 332 were expressed normally, including genes with a wide range of molecular functions. Compared with 20 different species of animals, microsatellites in the oyster genome typically exhibited 1) an intermediate overall frequency; 2) relatively uniform contents of(A)n and(C)n repeats and abundant long(C)n repeats(≥24 bp); 3) large average length of(AG)n repeats; and 4) scarcity of trinucleotide repeats. The microsatellite-fl anking regions exhibited a high degree of polymorphism with a heterozygosity rate of around 2.0%, but there was no correlation between heterozygosity and microsatellite abundance. A total of 19 462 polymorphic microsatellites were discovered, and dinucleotide repeats were the most active, with over 26% of loci found to harbor allelic variations. In all, 7 451 loci with high potential for marker development were identifi ed. Better knowledge of the microsatellites in the oyster genome will provide information for the future design of a wide range of molecular markers and contribute to further advancements in the fi eld of oyster genetics, particularly for molecular-based selection and breeding.     

Effects of Salinity on Embryonic Development,Survival, and Growth of Crassostrea hongkongensis

This study examined the effects of salinity on embryonic development,survival,and growth of the Hong Kong oyster Crassostrea hongkongensis.The embryos,larvae,and juveniles of C.hongkongensis were held in artificial seawater at three different salinities （low,15; medium,23; and high,30） to determine the optimum hatchery and nursery conditions for mass production of the seeds.Results showed that the percentage production of straight-hinged larvae from fertilized eggs was significantly lower at the high salinity than at the low-and medium-salinities （P ＜ 0.05）.The survival rates of larvae and juveniles differed significantly among the three salinity trials,with the highest survival rate observed at the low salinity （P＜ 0.05）.The shell height of larvae was significantly larger at the low salinity than at the high and medium salinities from days 9 to 15 （P＜ 0.05）,whereas that of juveniles was significantly larger at the low salinity than at the high and medium salinities on day 70 （P ＜ 0.05）.These results indicate that the larvae and juveniles of C.hongkongensis are tolerant to a wide range of salinities （15 to 30）,but show better growth and survival at relatively low salinities.Thus,it is recommended to use relatively low salinities in hatchery and nursery systems for improved yields of C.hongkongensis.     

不同规格近江牡蛎的排氨率和耗氧率分析

采用实验生态学方法研究了不同规格近江牡蛎的呼吸和排泄生理生态学参数。结果显示,近江牡蛎耗氧率R_O随软体干重(W )的增加而减小, 体重对近江牡蛎耗氧率(R_O)的影响达极显著水平,R_O = 1.8232W^-0.7600, (R²= 0.8313 , P <0.01); 近江牡蛎的排氨率(R_N)和排磷率(R_P)随软体干重的增加而降低,呈负相关关系;而氧氮比O/N和氮磷比N/P在各组间未达显著水平(P >0.05) , R_N = 0.0930W^-0.6582 (R² = 0.7444, P <0.01)、R_P = 0.0126W^-0.8874 (R² = 0.9224,P <0.01);各组不同规格近(大小)江牡蛎的O/N值较高,而N/P值则随规格增大而呈上升趋势,但差异也不显著(P >0.05)。这些研究结果对牡蛎养殖密度和环境关系分析及牡蛎健康养殖具有重要参考价值。     

Hauraki Gulf snapper fishery, 1972 and 1973: Some evidence for a declining catch‐rate

The commercial fishery for snapper Chrysophrys auratus (Bloch and Schneider) in the Hauraki Gulf is one of New Zealand's principal marine fisheries. The snapper catch from this area declined by 22% in 1972, and by a further 4% in 1973, following 4 y of high catches by Danish seiners.

Some of the decline can be attributed to economic factors within the industry, and adverse winter weather. However, the 1972 decline in Danish seine catches is an acceleration of an existing trend. It probably results from heavy exploitation of an accumulated stock, and from movement of a few strong year classes past the age at which they have contributed most to the catch. The slight recovery in the 1973 Danish seine catch resulted only from increased effort; catch per day at sea continued to fall. An improvement in catch and catch per day is unlikely for some years at least.

Snapper catches by other methods (trawl, long line, and set net) also declined during 1972, but the available data on fishing effort do not allow a detailed analysis. The trawl catch in the gulf fell further in 1973 as fishing effort was diverted to other grounds, but the catch by lines and nets improved slightly.     

Comparative descriptions of some oyster larvae from New Zealand and Chile,and a description of a new genus of oyster,Tiostrea

Detailed comparisons of the larvae of four species of New Zealand oyster (Saccostrea glomer‐ata, Crassostrea gigas, Ostrea lutaria, and a new species of Ostrea) and one Chilean oyster (Ostrea chilensis) showed that larvae of species other than O. lutaria and O. chilensis share certain features. One conspicuous feature is a distinct tract, termed here the posterior dorsal sulcus, which runs from the posterior margin of prodissoconch I to the valve margin of prodissoconch II; other characteristic features are an umbonate, inequivalve shell and a provinculum bearing distinct teeth. The larvae of O. lutaria and O. chilensis, in contrast, lack the posterior dorsal sulcus, are nonumbonate, and have nearly equal valves with an edentulous provinculum. These distinctive features, together with other adult features, suggest that the two species are more appropriately included in a separate genus. The name Tiostrea is proposed for the new genus. Certain supraspecific groupings within the Ostreinae need to be re‐examined in the light of breeding experiments.     

Larval development of Boccardia knoxi,a shell‐infesting spionid polychaete

Larvae of Boccardia knoxi (Rainer) were reared in the laboratory after dissection from brood capsules extracted from the outer shell and perio‐stracum of Cookia sulcata and Haliotis iris collected from Tasman Bay, New Zealand. All the eggs within each brood capsule developed into embryos. The dissected larvae had provisional setae and fed on phytoplankton. Larval development is described through to metamorphosis.     

长牡蛎(Crassostrea gigas)两个Dmrt家族基因的时空表达

本文以长牡蛎27个幼虫发育阶段个体以及成体的5个组织织作为实验材料,采用实时荧光定量PCR技术对其Dmrt家族中的2个基因(CgDsx和CgDmrtA2)的表达模式以及在性别决定中的作用进行了研究。结果表明,长牡蛎CgDsx基因在胚胎发育初期有大量表达,从囊胚期到担轮幼虫初期表达量最高,之后表达量开始降低,在D形幼虫后一直维持在极低的表达水平,此后仅在成体的雄性性腺中具有高度表达。由此可见,CgDsx可能也对早期胚胎发育起一定调控作用,同时参与了性别决定。CgDmrtA2在长牡蛎所有组织中均有表达,各组织间表达差异不显著,在D形幼虫至壳顶后期表达量较高,说明它参与了胚胎中后期的发育过程,可能与神经的形成相关,但是其具体功能还需要进一步研究。     

香港牡蛎(Crassostrea hongkongensis)水通道蛋白基因AQP1的克隆、分子特性和表达分析

水通道蛋白1(Aquaporin1,AQP1)是一类通过渗透梯度将水或一些小的中性分子快速穿过细胞膜的通道蛋白。通过RT-PCR和RACE技术克隆获得了香港牡蛎AQP1基因全长,并命名为Ch AQP1(Gen Bank登录号:KJ704847)。该基因全长1153bp,开放阅读框长度为888bp,编码295个氨基酸残基。Ch AQP1基因包括1个保守的MIP结构域、6个跨膜区、5个连接环、2个NPA(Asn-Pro-Ala)基序和选择性水孔构件ar/R(aromatic/arginine)。系统学分析表明Ch AQP1属于AQP1-like型水通道蛋白。m RNA组织分布结果显示,Ch AQP1在各个组织中均有表达,其中在闭壳肌和外套膜中表达量相对较高。利用实时定量PCR分析高、低盐胁迫下其在鳃中的表达模式,结果表明,Ch AQP1 m RNA表达量在低盐处理下基本没有太大变化;在高盐胁迫下,第1天(P0.01)、第3天和第5天(P0.05)显著下调;这说明Ch AQP1基因参与了香港牡蛎的渗透压平衡调节。     

长牡蛎(Crassostrea gigas)中ChIP-seq 方法的建立

染色质免疫共沉淀技术(ChIP)作为研究DNA 结合蛋白与DNA 相互作用最有力的工具, 越 来越受到重视。但目前该技术在牡蛎等贝类中的应用尚未见相关报道。为了研究长牡蛎早期胚胎发 育过程中表观遗传信息的变化模式, 本文在长牡蛎胚胎中探索并建立了一套完整的染色质免疫共沉 淀方法, 构建了高质量的高通量测序文库, 首次获得了牡蛎全基因组水平的组蛋白H3K4me3 修饰分 布, 从而为研究牡蛎的表观遗传调控奠定了基础。