Triploid and tetraploid Zhikong scallop, Chlamys farreri Jones et Preston, produced by inhibiting polar body I

Triploid scallops are valuable for aquaculture because of their enlarged adductor muscle, and tetraploids are important for the commercial production of triploids. We tested tetraploid induction in the zhikong scallop by inhibiting polar body I in newly fertilized eggs. The ploidy of resultant embryos was determined by chromosome counting at 2- to 4-cell stage and by flow cytometry thereafter. Embryos from the control groups were mostly diploids (79%), along with some aneuploids. Embryos from the treated groups were 13% diploids, 18% triploids, 26% tetraploids, 13% pentaploids, and 36% aneuploids. Tetraploids, pentaploids, and most aneuploids suffered heavy mortality during the first week and became undetectable among the larvae at day 14. Five tetraploids (2%) were found among a sample of 267 spat from one of the replicates, and none was detected at day 450. The adductor muscle of triploid scallops was 44% heavier (P < .01) than that of diploids, confirming the value of the triploid technology in this species.

Construction and characterization of two bacterial artificial chromosome libraries of Zhikong scallop, Chlamys farreri Jones et Preston, and identification of BAC clones containing the genes involved in its innate immune system

Large-insert bacterial artificial chromosome (BAC) libraries are necessary for advanced genetics and genomics research. To facilitate gene cloning and characterization, genome analysis, and physical mapping of scallop, two BAC libraries were constructed from nuclear DNA of Zhikong scallop, Chlamys farreri Jones et Preston. The libraries were constructed in the BamHI and MboI sites of the vector pECBAC1, respectively. The BamHI library consists of 73,728 clones, and approximately 99% of the clones contain scallop nuclear DNA inserts with an average size of 110 kb, covering 8.0x haploid genome equivalents. Similarly, the MboI library consists of 7680 clones, with an average insert of 145 kb and no insert-empty clones, thus providing a genome coverage of 1.1x. The combined libraries collectively contain a total of 81,408 BAC clones arrayed in 212 384-well microtiter plates, representing 9.1x haploid genome equivalents and having a probability of greater than 99% of discovering at least one positive clone with a single-copy sequence. High-density clone filters prepared from a subset of the two libraries were screened with nine pairs of Overgos designed from the cDNA or DNA sequences of six genes involved in the innate immune system of mollusks. Positive clones were identified for every gene, with an average of 5.3 BAC clones per gene probe. These results suggest that the two scallop BAC libraries provide useful tools for gene cloning, genome physical mapping, and large-scale sequencing in the species.

ICP-OES测定浮游有孔虫的Mg/Ca比值及其在SST分析中的应用

海洋是驱动大气环流的主要“引擎”，海表温度（SST）又是揭示海洋环境变化的重要参数。利用有孔虫Mg/Ca比值恢复SST具有较高精密度，是目前恢复SST较成功的方法之一。本论文旨在建立一套有效的有孔虫清洗方法和ICP-OES测试有孔虫Mg/Ca比值的分析方法。 文中分析了53份西菲律宾海区(N18°40.23′，E135°37.11′；水深3225m)表层沉积物中浮游有孔虫Globigerinoides ruber（白色）Mg/Ca比值。结合本研究区的特点，文中使用的清洗方法是有孔虫Mg/Ca比值测试常用的“Mg”清洗方法，主要是由以下步骤构成：①用超纯水和甲醇在超声波水浴中清洗，②显微镜下剔除深色硅酸盐颗粒，③用氧化试剂去除有机质(加入30%H2O2和0.1M NaOH溶液在90℃左右的沸水浴中加热)，④酸洗(250μl 0.001N HNO3 超声波水浴中清洗10S)。清洗干净的样品经超纯硝酸溶解成溶液后，利用电感耦合等离子体发射光谱(ICP-OES)对其进行Mg/Ca比值的分析测试。仪器操作条件如下：辅助气流量（Ar）为0.5L/min（0.5L/min-1.5L/min），雾化器压力为0.2Mpa（0-0.4Mpa），泵速为20rpm（0-125rpm），高频输出功率（RF）为1150W（750W-1500W），火焰高度为15.5mm（8-21mm），通过多次重复测量一组Mg/Ca=3.333mmol/mol的标准溶液，其Mg/Ca短期精密度<0.5%,长期精密度为1%。53份样品之间Mg/Ca比值的RSD为2.7%，利用Lea(2000)建立的太平洋地区浮游有孔虫G. ruber(白色)的Mg/Ca与SST的校正公式:Mg/Ca(mmol/mol) ＝0.30exp[0.089×SST(℃)]，得到本研究区的SST为28℃±0.3℃。

Growth, survival and immune activity of scallops, Chlamys farreri Jones et Preston, compared between suspended and bottom culture in Haizhou Bay, China

We examined the growth, survival and immune response of the scallop, Chlamys farreri, during a 1-year period in deep water of Haizhou Bay. Scallops were cultured using two methods: (1) in lantern nets at a 5 m depth and (2) in a bottom culture system (sleeves) on the seabed at about a 25 m depth. Shell heights, meat dry weight and immune activities in the haemolymph (superoxide dismutase and myeloperoxidase) were measured bimonthly or quarterly from July 2007 to June 2008. Survival was measured at the end of the study and environmental parameters in the experimental layers were monitored during the experiment. The growth and immune activities of scallops were lower when the water temperature was high, which was consistent with the main mortality occurring in summer. The growth and immunity of scallops were higher in the suspended culture than in the bottom culture during the experiment, with the exception of shell growth during the last study period. Survival of scallops in the suspended culture (54.6 +/- 12.3%) was significantly lower than that in the bottom culture (86.8 +/- 3.5%) at the end of this study. We conclude from our results that the high mortality of C. farreri can be prevented by culturing them in a bottom culture system before November of the first year, and then transferring them to a suspended culture to improve scallop production.

INVOLVEMENT OF CA(2+) SIGNALING PATHWAY DURING OOCYTE MATURATION OF THE NORTHERN QUAHOG MERCENARIA MERCENARIA

In many molluses, it has been found that Ca2+ signaling pathway is involved in the resumption of meiotic maturation in oocytes. To better understand the possible role of Ca2+ signaling pathway in regulating meiotic maturation in oocytes of the northern quahog Mercenaria mercenaria, free extracellular Ca2+, A23187 (calcium ionophore), verapamil (calcium channel blocker), and trifluoperazin (calmodulin antagonist) were used to incubate oocytes or serotonin-induced oocytes by pharmacological methods. Results show that extracellular Ca2+ (50 similar to 200 mM) and A23187 (1 similar to 10 mu M) can stimulate the meiotic maturation. In addition, verapamil (1 similar to 100 mu M) and trifluoperazin (10 similar to 1,000 mu M) could inhibit serotonin-induced oocyte maturation. Therefore, Ca2+ is essential for the reinitiation of meiotic maturation in oocytes of the northern quahog. Moreover, an increase i [Ca2+]i can promote meiotic maturation.

Studies on mass summer mortality of cultured zhikong scallops (Chlamys farreri Jones et Preston) in China

Mass mortalities of cultured zhikong scallops (Chlamys farreri) have occurred each summer in most culture areas of northern China since 1996. Among the hypothesized causes are high culture density, infectious disease and genetic inbreeding. To investigate these potential agents, C. farreri were deployed at three densities (low, medium and high) at three sites (Jiaonan, Penglai and Yantai) in the summer of 2000. Scallops were sampled for survival, growth and histopathology before, during and after a mortality episode. Most of the mortality occurred in July and August, during and toward the later part of the spawning season, when water temperature reached 23-26 degrees C. Final cumulative mortalities reached 85% to 90% at all three sites. Scallops in the medium and high densities had higher initial death rates than did those at the low density. High densities also inhibited growth. Ciliates from the genus Trichodina, larvae of various organisms and anomalous secretions were observed in sections of the gill cavity, with highest prevalence during and at the end of the mortality period. Prokaryotic inclusion bodies were found in the soft tissues, but their prevalence was low and apparently without correlation with mortalities. Genetic analysis with random amplified polymorphic DNA markers showed slightly lower heterozygosity in the cultured stocks (0.301) than in the wild stocks (0.331). It is possible that the mortalities are caused by a combination of several factors such as stress associated with reproduction, high temperature, overcrowding and poor circulation in the growout cages, opportunistic invaders or pathogens, and possibly inbreeding. (c) 2005 Elsevier B.V. All rights reserved.

低氧对高原鼠兔肝细胞内质网和心肌肌浆网Ca~(2+)泵的影响

模拟高原低氧条件研究高原鼠兔肝细胞内质网(Endoplasmic reticulum , ER ) 和心肌肌浆网(Sarcoplasmic reticulum , SR) 钙泵功能变化。实验设对照组(海拔2 300 m ) 和两个低氧实验组(模拟海拔5 000m 和7 000m )。24 h 急性低氧时, 海拔5 000 m 组高原鼠兔ER 的Ca2+ 泵活性无变化, 海拔7 000 m 组高原鼠兔ER Ca2+泵活性下降29102%。7 d 亚急性低氧时高原鼠兔SR 的Ca2+泵活性无显著变化。高原鼠兔ER 的Ca2+泵活性在海拔5 000 m 组和7 000 m 组分别升高32.50% 和33.33%。25 d 慢性低氧时高原鼠兔ER , SR 的Ca2+ 泵活性均无显著变化.表明: 急性低氧对Ca2+泵功能有抑制作用, 低氧7 d 后抑制缓解, 至25 d 低氧时趋于恢复。

水热条件下钾长石碱性分解反应机理的实验研究

It is well known that our country is short of water-soluble potassium, but rich in insoluble potassium ores. Based on the work of the formers, using the orthogonal and monofactor experiments, the author optimized the production technology of micro-porous potassium silicon calcium mineral fertilizer by non-stirring hydrothermal chemical reaction when the alkaline earth booster CaO was available. The influences of temperature、time、reactant ratio and water-solid ratio on the dissolution rate of production’s elements were studied by orthogonal experiments, and the production technology was further optimized by monofactor experiments. By XRD、SEM、EDS and dissolving experiments, it was systematically studied that the effects of the reactant ratio、reaction time and reaction temperature on the properties of the production obtained by the hydrothermal reaction between KAlSi3O8 and CaO. The results showed that：when changing of the reaction condition, the reaction productions included tobermorite、 hibschite、α-C2SH and K2Ca(CO3)2; among which, K2Ca(CO3)2 was not the first production containing potassium, but K2Ca(CO3)2 was synthesized by the reaction among KOH、Ca(OH)2 and CO2. Whether the phase was synthesized was related to not only the reaction condition, but also their physicochemical properties; when the reaction condition was changed, the changes of different phases were different. The results of XRD and dissolution rate experiments explained the dissolution characteristic of every element of hydrothermal productions very well, and the relation between the dissolution rate of element and the phase of productions poured a good illumination on the production technology. The results of SEM and EDS showed that: hydrogarnet looked like spherical, and its surface was covered by some productions including K phase and Ca、Si phase; but the morphology of tobermorite was platy or lamellar or needlelike, and parts of Si in the structure of tobermorite were substituted by Al，and some K+ cations were inserted into the Ca interlayer of tobermorite at the same time. It was the first time that the interface between KAlSi3O8 and Ca(OH)2 was observed directly by SEM and EDS after the hydrothermal reaction, and the mechanism of hydrothermal reaction of KAlSi3O8 and Ca(OH)2 was further discussed. These results indicated that: the Ca-KAlSi3O8 intermediate compound was formed at first, and some K was released into the solution and KOH was produced at the same time; the C-S-H phase appeared before hydrogarnet, and then hydrogarnet was synthesized when the chemical reaction was carried on; if the reaction was carried on furthermore, α-C2SH、tobermorite and other C-S-H phases of different atom ratio appeared. The author found that the structure of KAlSi3O8 would be more drastically destroyed if there were some reactants, such as Ca(OH)2 which reacted with KAlSi3O8 and new phases were formed after the hydrothermal reaction between KAlSi3O8 and alkaline solution of equal ionic strength was finished. With the combination of calcination and hydrothermal reaction methods, the dissolution rate of products were greatly improved when the hydrothermal reaction was carried out after KAlSi3O8 and CaCO3 were calcined. Furthermore, the author has tentatively explored how to evaluate the effects of the differences of the activity of lime on the dissolution properties of hydrothermal products.

生物壳体Sr／Ca、Mg／Ca、^87Sr／^86Sr比值在恢复古气候、古环境中的应用—以小渡口剖面为例

生物壳体的Sr/Ca、Mg/Ca比值能反映其沉积水体的Sr/Ca、Mg/Ca比值，在一定条件下反映水体的盐度、温度，并分别与宿生水体的盐度、温度呈正比关系。生物壳体的^87Sr/^86Sr比值是恢复盆地古水文条件的一种重要手段。本文以著名的泥河湾盆地小渡口剖面的第28层为实例，力图通过对有孔虫、介形虫、腹足类生物壳体Sr/Ca、Mg/Ca比值与^87Sr/^86Sr比值的对比研究，并结合前人所作的中更新世古地理特征研究，来恢复生物壳体沉积时泥河古湖水的温度、盐度，进而进行古气候、古环境的恢复。研究结果表明：在0.97-0.94Ma间，古泥河湾湖为一陆相湖泊，湖水的温度、盐度变化趋势相吻合，均呈低－高－低的演化特征，可能对应着盆地水体的扩展－退缩－扩展变化。其中，在0.96-0.95Ma间，古泥河湾湖具有较高的温度、盐度特征。生物壳体的Sr/Ca、Mg/Ca比值能反映其沉积水体的Sr/Ca、Mg/Ca比值，在一定条件下反映水体的盐度、温度，并分别与宿生水体的盐度、温度呈正比关系。生物壳体的^87Sr/^86Sr比值是恢复盆地古水文条件的一种重要手段。本文以著名的泥河湾盆地小渡口剖面的第28层为实例，力图通过对有孔虫、介形虫、腹足类生物壳体Sr/Ca、Mg/Ca比值与^87Sr/^86Sr比值的对比研究，并结合前人所作的中更新世古地理特征研究，来恢复生物壳体沉积时泥河古湖水的温度、盐度，进而进行古气候、古环境的恢复。研究结果表明：在0.97-0.94Ma间，古泥河湾湖为一陆相湖泊，湖水的温度、盐度变化趋势相吻合，均呈低－高－低的演化特征，可能对应着盆地水体的扩展－退缩－扩展变化。其中，在0.96-0.95Ma间，古泥河湾湖具有较高的温度、盐度特征。

喀斯特石灰土-灌丛系统主要养分含量分布特征研究---以贵阳花溪杨中石灰土-灌丛系统Ca、Mg、 K 、P为例

本论文通过对贵州贵阳花溪杨中喀斯特山区的植被调查与土壤样品分析, 研究土壤-植被中Ca、Mg、 K、P分布特征、变化规律及其影响因素，并在此基础上尝试Ca同位素的预处理研究，以期为喀斯特地区土壤-植被中Ca、Mg、 K、P（尤其是Ca）的生物地球化学循环研究奠定基础，同时为植被演替、物质循环研究和退耕还林还草的人工恢复提供植物自身的营养依据。本次研究取得如下认识： 1.受石灰岩母岩控制，土壤样品元素含量的特征为：CaO>MgO>K2O> P2O5。同一元素沿海拔变化有一定的相关性，并且相关性随元素的不同存在差异：随海拔高度的增加CaO含量有升高的趋势，而MgO 、K2O、Na2O含量都有所降低。表层土壤中K2O-Na2O含量正相关变化，CaO-K2O含量负相关变化。并且从土壤表层到底层在剖面-1中随深度增加，CaO、MgO、K2O含量升高，而P2O5的含量则有上升的趋势。土壤中元素的上述分布特征主要与石灰岩的风化成土作用过程中不同分化程度土壤Ca的淋失和来自高处的富钙雨水输入有关。 2.喀斯特生态环境富钙的这一特点对喀斯特灌丛植物的种类成分和营养元素含量有很大影响。地貌部位、海拔高度对植物Ca、Mg 、K、P含量也有不同程度的影响。植物Ca、Mg 、K、P含量由高到低依次为：Ca > K > Mg >P。苔藓与其它植物比较， Ca、Mg、 K、P的含量都小于其它三类较高等的植物，这一现象与其自身的生物学特性有关：苔藓植物组织结构简单，不具备发达的输导组织，不具备真正的根系，不利于从土壤等基质中吸收营养。在、灌草、灌木、藤本三类植物营养元素之间Ca –P含量具有负相关变化，Mg –P含量具有正相关变化；在所分析的植物种，P- K含量变化呈正相关，并且植物叶片Ca/K比值随着P含量的增加而显著地以指数形式减小。这是因为植物体内的K和P对于Ca来说起着平衡离子（counter ion）的作用。 3. 生境中基岩裸露和土体浅薄的特点，极大地制约了喀斯特灌丛的发育，喀斯特生境的富含钙的特点对喀斯特灌丛植物的种类成分和元素含量有很大的影响。但是杨中灌丛群落植物Ca、Mg、K、P含量和对应土壤CaO、MgO、K2O和P2O5含量相关性不明显。植物Ca、Mg、K、P元素虽主要来源于土壤，但因其含量受元素地球化学性质、植物种类、土壤元素对植物有效态的含量等因素的制约，而使植物元素含量和土壤元素的含量没有明显的相关性． 4.从Ca同位素的预处理研究来看，离子交换分离法对去除样品中的杂质效果较好。在以后测定过程中，有利于增强离子流稳定性和强度, 提高测量精度和准确度，与沉淀法相比较，此法适宜作为钙同位素样品的前处理方法。 关键词： 喀斯特 石灰土-灌丛系统 Ca、Mg、K、P含量分布 钙同位素 环境地球化学

湖泊沉积物碳氧同位素和介形虫Li/Ca比值与古环境重建

了解过去是认识现在和预测未来的基础。晚全新世是PAGES研究的目标时段之一。寻找和解译最近2000年连续、高分辨率自然记录是古气候研究的热点与难点。本项研究有针对性地选择程海、草海和青海湖开展湖泊沉积物环境记录的精细研究，结合14C、210Pb和137Cs定年，系统探讨了碳酸盐碳氧同位素、纤维素碳氧同位素、有机质碳同位素以及介形虫壳体Li/Ca比值指标的古环境指示意义。 通过研究，本论文取得了如下成果和新的认识： 1. 建立了一套有效的四阶段湖泊沉积物有机质纤维素提取方法（5%NaOH碱洗、5%HCl酸洗、亚氯酸钠和冰醋酸混合溶液漂白以及17.5%NaOH碱洗），红外光谱鉴定提取物为纯的α-纤维素，表明该实验方法是可行的，为今后广泛开展湖泊沉积物纤维素稳定同位素研究打下了坚实基础。 2. 多指标综合辨识了程海和草海沉积物碳酸盐主要是自生碳酸盐。程海和草海沉积物有机质C/N比值结合有机质碳同位素结果表明两湖的有机质分别源于水生植物藻类和大型水草。程海和草海沉积物碳酸盐含量主要反映了湖区温度的变化。 3. 程海、草海沉积物碳酸盐与青海湖沉积物介形虫壳体氧同位素组成均反映了湖区降水/蒸发比。降水/蒸发比大的湿润期，碳酸盐δ18O值小；降水/蒸发比小的干旱期，碳酸盐δ18O值大。程海沉积物碳酸盐碳同位素组成影响因素复杂，除了受大气与湖水之间的CO2交换影响外，还受水生植物光合/吸收作用的影响；草海沉积物碳酸盐碳同位素组成更大的变化范围，反映了湖区水生植物光合/吸收作用的影响，其异常的正值可能指示了湖区细菌参与有机质碳同位素分馏过程。 4. 利用草海沉积物有机质纤维素氧同位素定量恢复了湖水氧同位素组成变化。在此基础上，结合碳酸盐氧同位素组成初步恢复了草海地区过去500年来温度变化历史：草海地区在过去500年明显存在四个冷期，包括1550－1610年，1670－1730年，1770－1870年和1890－1920年冷期，其中前三个发生在传统意义上的现代小冰期时段。与其它记录研究结果的一致性表明纤维素氧同位素结合碳酸盐氧同位素是恢复古温度变化的最有效途径之一，同时也为现代小冰期在中国西南地区的存在提供了湖泊沉积学方面的证据。 5. 不同类型水生植物湖泊，湖泊沉积物有机质δ13C值对湖泊初级生产力变化的响应过程不同。大型水草为主的湖泊（草海），其沉积物有机质δ13C值随湖泊生产力的增大呈现增加的变化趋势；藻类为主的湖泊（程海），其沉积物有机质δ13C随湖泊生产力的增大呈现减小的变化趋势，藻类易降解是导致δ13C值随湖泊生产力的增大呈现减小变化趋势的主要原因。 6. 程海沉积物碳酸盐和有机质碳同位素组成的正相关变化以及草海沉积物碳酸盐和有机质碳同位素组成的负相关变化.表明湖泊生产力变化并不是导致碳酸盐和有机质碳同位素正相关变化的主要原因，湖泊水生植物类型以及湖泊大小均起着重要作用。湖泊沉积物碳酸盐与有机质之间的碳同位素分馏（△δ13C）是一种有效的湖泊生产力指示剂，即使是在有微生物参与有机质碳同位素分馏过程的草海，△δ13C值也反映了湖泊生产力的变化过程。 7. 青海湖沉积物单一种属介形虫壳体胖真星介（Eucypris inflata）Li/Ca比值与气象记录以及邻近地区都兰和祁连山树轮宽度指数恢复的古温度变化序列的对比研究揭示，介形虫壳体Li/Ca比值与温度呈明显的负相关变化（Li/Ca比值高，温度低；Li/Ca比值低，温度高），表明介形虫壳体Li/Ca比值是一种有效的古温度代用指标。 8. 青海湖沉积物单一种属介形虫壳体胖真星介（Eucypris inflata）Li/Ca比值、氧同位素与反映太阳活动的大气14C含量和冰芯10Be含量的一致性变化表明青海湖地区温度和降雨量的同步变化主要受太阳活动控制。