新型皱纹盘鲍防御素hd-def的cDNA序列分析、基因组克隆及表达研究

从构建的皱纹盘鲍肝肾cDNA文库中筛选到了鲍防御素基因EST.通过序列分析发现该基因的全长cDNA序列编码66个氨基酸残基,其前体由信号肽、前导肽和成熟肽组成.该前体的成熟肽含42个氨基酸(6个Cys),推测分子量为4323Da,等电点为8.02.氨基酸序列同源性分析表明,该多肽与昆虫防御素的相似性较高,最高可达70%.因成熟肽二级结构具有典型的昆虫防御素结构特征,该多肽应属于抗菌肽中的昆虫防御素亚家族,是一种新型抗菌肽,将其命名为鲍防御素hd-def.采用基因组步移法获得了全长4032bp的基因组序列.分析表明,该基因由3个内含子和4个外显子编码组成;3个内含子大小分别为497bp、2357bp和528bp,其中两个内含子存在于编码信号肽的序列中.用鳗弧菌和金黄色葡萄球菌刺激皱纹盘鲍,能诱导hd-def的表达.实验检测了5种组织,发现hd-def基因仅在肝胰腺中表达,具有明显的组织表达特异性;其表达属于诱导型表达,提示该基因可能参与皱纹盘鲍的抗细菌感染.     

维生素D对皱纹盘鲍生长和体组织抗氧化反应的影响

设计了维生素D添加水平分别为0IU/kg(缺乏组),5×103 IU/kg(正常组)和5×105 IU/kg(过量组)的3种半精制饲料,在循环水养殖系统中用其饲养初始体重为(0.81±0.00)g,初始壳长为(16.42±0.03)mm的皱纹盘鲍240d,以期探讨维生素D对皱纹盘鲍生长和体组织抗氧化反应的影响.结果表明:与维生素D正常添加组相比,维生素D缺乏对皱纹盘鲍的增重率(WGR)和贝壳日增长(DISL)均没有显著差异(P＞0.05);维生素D过量则显著降低皱纹盘鲍的增重率和贝壳日增长(P＜0.05).皱纹盘鲍的不同组织(内脏团和肌肉)对维生素D缺乏或过量的敏感性不一致.维生素D缺乏显著降低内脏团中的GPX(谷胱甘肽过氧化物酶)和GST(谷胱甘肽-S转移酶)活力(P＜0.05),显著提高GR(谷胱甘肽还原酶)/GPX比值(P＜0.05),对CAT(过氧化氢酶)、SOD(超氧化物歧化酶)、GR活力以及CAT/SOD比值没有显著影响(P＞0.05);而维生素D过量显著降低SOD和CAT活力的同时(P＜0.05),显著升高GPX和GR活力(P＜0.05).肌肉中,维生素D缺乏对CAT、SOD、GST和CAT/SOD比值没有显著影响(P＞0.05),但显著降低GPX和GR活力(P＜0.05),同时显著升高GR/GPX比值(P＜0.05);维生素D过量对CAT、GPX、GST、CAT/SOD和GR/GPX比值没有显著影响(P＞0.05),但显著升高SOD和GR活力(P＜0.05).     

缺磷对皱纹盘鲍贝壳生物矿化的影响

以皱纹盘鲍（Haliotis discus hannai Ino）成鲍为对象，采用“平珠”技术，将圆形玻片插入鲍鱼的外套膜外腔，配制了2种不同磷含量的半精制饲料，在缺磷的人工海水中喂养鲍鱼40天。养殖结束后取出“平珠”，利用扫描电镜对其超微结构进行了观察，并与正常贝壳作比较。结果发现：磷添加饲料组所得“平珠”具有与正常贝壳一致的矿化结构，而缺磷饲料组所得“平珠”晶体的形成、生长和结构都受到显著影响，方解石的排列失去规律，文石的晶态也变得无规则。本研究证明了“平珠”生成法能够模拟贝壳的自然生长，是研究贝壳生物矿化及其营养学机理的有效途径。同时证明了磷对于维持贝壳的晶体形成、正常生长和结构是必不可少的。     

马氏珠母贝选育系遗传变异的AFLP分析

利用AFLP技术对马氏珠母贝两个选育系DDS-G1和JCS-G1及大亚湾养殖群体(DDC)进行了遗传变异分析.5对选择性扩增引物共扩增谱带265条,DDC的多态位点比率为79.25%,DDS-G1为73.58%,JCS-G1为75.85%.两个选育系的单态位点明显增加,而Shannon's遗传多样性指数和Nei's基因多样性有所下降.三个组间的遗传距离为0.1615～0.2744,两个选育系间的遗传距离较大.研究结果表明,经过一代的选择,马氏珠母贝选育系的遗传结构和遗传多样性已发生改变,AFLP标记能够有效地监测选育系的遗传变异.     

獐子岛海洋生态牧场

正獐子岛海洋牧场位于黄海北部北纬39度海域。经过几十年的建设和创新发展,构建了以虾夷扇贝、海参、皱纹盘鲍、海胆、海螺、三倍体单体牡蛎等海珍品增养殖为主、大型海藻生态调控为辅的生态系统水平的海洋牧场,并正在完善育种-育苗-养殖-增殖-加工全产业链的海洋牧场生态产业体系。     

AFLP技术在水生动物遗传学研究中的应用及前景展望

综述了AFLP技术在水生动物遗传多样性、亲子代遗传关系、遗传图谱构建及功能基因相关分子标记、主效基因筛选等方面的应用，并简要介绍了AFLP技术发展及其在水生动物遗传学研究中的应用前景。     

五个紫菜品系间遗传差异的RAPD分析

应用随机引物扩增片段多态性（RAPD）技术对2种紫菜的5个品系进行遗传多样性分析。共筛选出21条随机引物,PCR反应得到147条扩增片段。根据共享扩增片段计算遗传相似性指数（F）和相对遗传距离,利用NJ法构建系统树。结果表明,条斑紫菜或坛紫菜的养殖品系首先聚类在一起,两个条斑紫菜养殖品系之间的遗传距离是0．32,两个坛紫菜养殖品系之间的遗传距离是0．31。条斑紫菜养殖品系CPY1－08A与坛紫菜养殖品系CPH8－83之间的遗传距离最大,达0．42。本文结果显示RAPD可以作为简便有效的分子工具应用于紫菜的遗传多样性和种质鉴定研究中。     

利用AFLP技术分析中国明对虾的韩国南海种群和养殖群体的遗传差异

利用AFLP技术对新发现的中国明对虾的一个地理种群——韩国南海种群（SP）和中国明对虾的养殖群体（CP）进行了遗传分析。每个群体随机取样30个,5对AFLP引物获得326个位点。其中SP多态位点比例（P0.99）46.93％,Shannon多样性指数（Ⅰ）0．1884,Nei（1978）基因多样性指数（h）0．1197,群体差异性位点9个,占检测位点总数的2．7％。CP多态位点比例（R9q）51．84％,Shannon多样性指数（Ⅰ）0．1954,Nei（1978）基因多样性指数（h）0．1229,群体差异性位点19个,占检测位点总数的5．8％。SP种群各项遗传参数都低干CP种群。两个群体的非偏差遗传相似度和遗传距离分别为0．9899和0．0102。利用中国明对虾的韩国南海种群和养殖群体杂交可以获得新的种质资源,这将为获得最大变异数量性状表型和基因多样性的产生提供可能。     

用RAPD技术对我国栉孔扇贝野生种群与养殖群体的遗传结构及其遗传分化的研究

利用RAPD技术对我国栉孔扇贝野生种群和养殖群体的遗传结构及其分化进行了研究，在对20个野生栉孔扇贝和20个养殖栉孔扇贝的基因组DNA的检测中，20个10碱基的随机引物共扩增出153条清晰可分辨的DNA片段，片段的长度为200－3000bp其中多态性片段分别为116和112条，野生种群和养殖群体的多态位点比例分别为75．82％，和73．47％，杂合度分别为0．27和0．26。根据基因频率计算出两个群体的近交系数为0．00129，遗传距离为0．028。栉孔扇贝野生种群的多态位点比例和杂合度处于较高水平，说明我国栉孔扇贝野生种群的遗传多样性水平较高，种质资源尚处于较好状态，应制定相应的渔业生产和管理措施加以保护，养殖群体的多态位点比例和杂合度都低于野生种群，这与人工累代养殖过程中，群体较小，近交机会增加有关。     

中国不同纬度野生大豆和栽培大豆AFLP分析

采用ＡＦＬＰ分子标记技术,对我国不同纬度的２０份野生大豆和载培大豆（Ｇ．ｍａｘ）进行了多样性分析,结果表明：（１）我国野生大豆的遗传多样性较栽培大豆更为丰富;（２）根据ＡＦＬＰ分析结果,将野生大豆和栽培大豆完全划分为２类,并发现野生大豆栽培大豆的种特异谱带,说明野生大豆和栽培大豆作为２个种有遗传基因的;（３）野生大啼和栽培大豆不同纬度品种间纬度相傧首先聚在一起,表明不同进化类型的大豆其遗传距离与纬     

16S rRNA基因与16S-23S rRNA转录单元内间隔区序列分析及其在节旋藻和螺旋藻分类鉴定中的应用

测定了节旋藻属3个品系和螺旋藻属1个品系的全长16SrRNA基因基因和16S rRNA转录单元内间隔区序列（ITS）,分析了已知的节旋藻、螺旋藻和相关品系的相应序列的同源性,构建了系统发生树,并评价了这两段DNA序列在节旋藻、螺旋藻种属分类和种质鉴定中的意义。结果表明：（1）16SrRNA基因序列和ITS序列均可用于节旋藻属和螺旋藻属的属间分类,以两序列为基础的系统学分析结果一致;（2）ITS序列变异程度高于16SrRNA序列,适用于节旋藻和螺旋藻属内品系或种质鉴定;（3）节旋藻属可明确界定,16SrRNA基因序列相似性大于98％,ITS序列相 似性大于88％;（4）螺旋藻属某些品系间16SrRNA序列和ITS序列相似性较低,与不同属间的序列相似性程度为同一水平。     

利用AFLP技术研究海湾扇贝不同养殖群体的遗传结构及其分化

采用AFIP分子标记技术对我国海湾扇贝的4个不同地理群体共80个个体利用7对引物组合进行了遗传多样性分析。4群体的多态位点比例分别为：加拿大养殖群体48．8235％,墨西哥湾养殖群体49．2914％,秦皇岛养殖群体38．2353％,浙江养殖群体41．1765％。平均杂和度分别为0．1878,0．1886,0．1265和0．1618。遗传距离介于0．1188～0．0941之间。4个群体的Fst为0．1883。根据遗传距离绘制了UPGMA聚类图。4个群体间的遗传关系如下：加拿大群体和浙江群体聚为一支,墨西哥湾群体和秦皇岛群体聚成一支,最后这两支聚在一起。试验结果表明,经过长时间累代养殖的浙江、加拿大、墨西哥湾群体仍保持较高的杂合度,与引种时间最短的(2年)、最为接近美国自然群体的秦皇岛群体比较,以上3个群体没有出现多态位点比例和杂合度显著下降的现象。     

Mechanical properties and structure of Strombus gigas,Tridacna gigas,and Haliotis rufescens sea shells: A comparative study

Sea shells are composed of calcium carbonate crystals interleaved with layers of viscoelastic proteins, having dense, tailored structures that yield excellent mechanical properties. Shells such as conch (Strombus gigas), giant clam (Tridacna gigas), and red abalone (Haliotis rufescens) have hierarchical architectures that differ depending on growth requirements and shell formation of the particular mollusk. Mechanical tests have been carried out on these shells for a comparison of strength with respect to the microstructural architecture and sample orientation. The mechanical response is found to vary significantly from specimen to specimen and requires the application of Weibull statistics in order to be quantitatively evaluated. The complex micro-laminate structure of these biocomposite materials is characterized and related to their mechanical properties. The red abalone has the highest compressive (233–540 MPa) and flexure strengths of the three shells. The giant clam has the lowest strength (87–123 MPa) and the conch has an intermediate value (166–218 MPa) in compression. The high compressive strength observed in the abalone is attributed to an optimization of microstructural architecture in the form of 2-D laminates, enhancing the fracture toughness of this shell material and enabling higher stresses to develop before fracture.     

Surface plasmon resonance imaging measurements of DNA and RNA hybridization adsorption onto DNA microarrays

Surface plasmon resonance (SPR) imaging is a surface-sensitive spectroscopic technique for measuring interactions between unlabeled biological molecules with arrays of surface-bound species. In this paper, SPR imaging is used to quantitatively detect the hybridization adsorption of short (18-base) unlabeled DNA oligonucleotides at low concentration, as well as, for the first time, the hybridization adsorption of unlabeled RNA oligonucleotides and larger 16S ribosomal RNA (rRNA) isolated from the microbe Escherichia coli onto a DNA array. For the hybridization adsorption of both DNA and RNA oligonucleotides, a detection limit of 10 nM is reported; for large (1,500-base) 16S rRNA molecules, concentrations as low as 2 nM are detected. The covalent attachment of thiol-DNA probes to the gold surface leads to high surface probe density (10(12) molecules/cm2) and excellent probe stability that enables more than 25 cycles of hybridization and denaturing without loss in signal or specificity. Fresnel calculations are used to show that changes in percent reflectivity as measured by SPR imaging are linear with respect to surface coverage of adsorbed DNA oligonucleotides. Data from SPR imaging is used to construct a quantitative adsorption isotherm of the hybridization adsorption on a surface. DNA and RNA 18-mer oligonucleotide hybridization adsorption is found to follow a Langmuir isotherm with an adsorption coefficient of 1.8 x 10(7) M(-1).     

Growth and structure in abalone shell

The growth and self-assembly of aragonitic calcium carbonate found in the shell of abalone (Haliotis) is described. This was accomplished through the close examination of laboratory-grown flat pearl samples and cross-sectional slices of the nacreous shell. Further understanding of the sequenced assembly has been obtained. It has been confirmed that the growth of the aragonite component of the composite occurs by the successive nucleation of aragonite crystals and their arrest by means of a protein-mediated mechanism; it takes place in the “Christmas-tree pattern” [Nature 49 (1994) 371]. It is shown that the protein layer is virtually absent where plates on a same plane abut (along lateral surfaces of tiles). This suggests a mechanism of c-axis aragonite growth arrest by the deposition of a protein layer of approximately 20–30 nm that is periodically activated and determines the thickness of the aragonite platelets, which are remarkably constant (0.5 μm). This platelet size was measured for animals with shell diameters of 10, 50, and 200 mm and was found to be constant. The overall growth process is expressed in terms of parameters incorporating the anisotropy of growth velocity in aragonite (V_c, the velocity along c axis, and V_ab, the velocity in basal plane). Comparison of laboratory-raised and naturally-grown abalone indicates growth regulated by the level of proteinaceous saturation. Naturally-grown abalone exhibits mesolayers (growth bands) 0.3 mm apart; it is proposed that they result from seasonal interruptions in feeding patterns, creating thicker (10–20 μm) layers of protein. These mesolayers play a critical role in the mechanical properties, and are powerful crack deflectors. The viscoplastic deformation of the organic inter-tile layers is responsible for the significant improvement of tensile strength over the tensile strength of monolithic aragonite.     

鲍鱼壳的结构和力学性能

以皱纹盘鲍鱼壳为对象,研究了其结构与力学性能之间的关系.研究表明,鲍鱼壳主要由方解石和文石构成,且外层为方解石,内层为文石.鲍鱼壳的力学性能随其加热温度的升高而明显降低,密度也呈现同样的变化,说明鲍鱼壳中所含有的少量有机质随着加热温度的升高而减少,致使鲍鱼的力学性能明显下降.     

Changes in the quality of abalone (Haliotis asinina Linnaeus) packaged under atmospheric air,vacuum and modified atmosphere

Changes in the quality of abalone (Haliotis asinina Linnaeus) meat packaged under modified atmosphere (MA) of 40% CO2/30% O2/30% N2, vacuum and atmospheric air in polyvinylidene chloride (PVDC)/nylon/oriented polypropylene (OPP) pouches, and stored at 2 ± 1°C were investigated. Biochemical indices, such as pH, total volatile basic nitrogen (TVB‐N), and trimethylamine (TMA) and sensory evaluation, as well as total plate counts of packaged abalone meat were determined periodically. TMA was not affected by the packaging conditions, and remained low during storage periods. A decrease in pH of the MA packaged abalones during the storage reflected the apparent absorption of CO2. The content of TVB‐N slowly increased in the MA packaged abalone, whereas those stored in atmospheric air markedly increased during the storage. The sensory quality of MA packaged abalones was shown to be acceptable up to 15 days, while atmospheric and vacuum‐packaged abalones were not acceptable after 3 days of storage. Copyright © 2007 John Wiley & Sons, Ltd.