拟穴青蟹GRP78基因的克隆与应激表达

葡萄糖调节蛋白78(glucose regulated protein 78 ku,GRP78)是热休克蛋白70家族成员之一,在调节蛋白质折叠和维持内质网稳态过程中起着分子伴 侣作用。采用RT-PCR、RACE等技术,首次从拟穴青蟹获得了GRP78的cDNA全长序列。该序列全长2 284 bp,开放阅读框(ORF)为1 962 bp,编码653个氨基酸残基。 同源分析显示,该基因编码的蛋白含有HSP70家族的签名序列,C末端为内质网蛋白滞留信号KDEL,与其他物种具有很高相似性。实时荧光定量PCR结果表明,GRP78 基因在拟穴青蟹多个组织中均有表达。第一期仔蟹在不同的温度和盐度下暴露12 h后,GRP78基因表达量随环境温度升高而增加;在高盐(30)条件下GRP78表达量 较高,进而推测拟穴青蟹GRP78参与蛋白质折叠和环境胁迫的应答。     

锯缘青蟹促雄腺发育的组织学研究

采用组织切片技术，以OlympusBH-2型显微镜对锯缘青蟹(Scylla serrata)促雄腺的发育进行观察。结果显示，促雄腺附着于射精管近末端，发育过程可分为4期：Ⅰ期促雄腺短小，腺细胞数量少；Ⅱ期促雄腺呈明显的索状；Ⅲ期促雄腺体积达到最大，一些部位膨大增生；Ⅳ期促雄腺不再生长，腺体急剧退化。腺细胞具有2种类型：A型细胞，核圆、异染色质少、胞质染色浅；B型细胞，核扁平、异染色质多，细胞染色深、界限不清晰，有时细胞质消失。促雄腺发育过程中，A、B型细胞数量、比例发生变化。B型细胞为A型细胞完成分泌后的存在形式，B型细胞比例反映了促雄腺分泌活动的状况。锯缘青蟹促雄腺发育和精巢成熟具有一定的相关性。     

江蓠与新对虾、青蟹混养试验

细基江蓠繁枝变种与新对虾，青蟹混养试验结果表明：（１）细基江蓠繁枝变种生长的最适温，盐度分别为１７－３０℃和１７－２９‰，这一温，盐度范围也适合新对虾和青蟹的生长；鱼塘海水的温，盐度变化与沿海不同，因此，移植野生江蓠到鱼塘养殖时要考虑这一特点。（２）江蓠在不同的生长季节的最适生长水层是不同的。（３）江蓠的主要有效氮是ＮＨ４＾＋，而新对虾，青蟹的排泄物可分解出大量的ＮＨ４＾＋。这样，江蓠与新对虾，青     

锯缘青蟹死亡的组织病理学初步观察

对2008年8月福建省泉州市某海水场养殖锯缘青蟹中发生的死亡现象,开展锯缘青蟹的组织病理学研究。解剖可见肌肉明显坏死、溶解;头胸甲储积大量暗黄色体液,经福尔马林固定成固态物质;光镜下发现病蟹的中肠腺上皮细胞坏死、细胞质中出现大型的包涵体;电镜下发现病蟹的细胞质中有长约为500hm、宽约为200hm杆状结构的疑似杆状病毒存在。     

A reovirus disease in cultured mud crab, Scylla serrata, in southern China

A reovirus, designated mud crab reovirus (MCRV), associated with large economic losses was recently isolated from marine cultured mud crab, Scylla serrata, in southern China. The complete viral particle is 70 nm in diameter, icosahedral and non-enveloped. The virus infects connective tissue cells of the hepatopancreas, gills and intestine in mud crab and develops in the cytoplasm. Hundred per cent mortality was observed in mud crab experimentally infected by intramuscular injection, bath inoculation and oral inoculation, while cohabitation infection caused 80% mortality. The viral genome consists of 13 linear dsRNA segments, with an electrophoretic pattern 1/5/7. The results of this study suggest that the virus is highly pathogenic and can be transmitted enterically as well as via the body surface of mud crab. Although the genomic organization of this virus is different from that of the other crab reoviruses, CcRV-W2 and DpPV, all three of these reoviruses have similar electrophoresis patterns. Therefore, MCRV may be a new member of the DpPV and CcRV-W2 group.     

拟穴青蟹Cactus基因的cDNA克隆、序列及生物学功能

为获取拟穴青蟹Cactus基因cDNA全长、分析基本生物学信息,并初步探索其在病原物刺激下的免疫反应,实验采用RACE技术获得了拟穴青蟹Cactus(SpCactus)基因的cDNA全长序列,其cDNA全长为2035 bp,开放阅读框(ORF)1311 bp,编码436个氨基酸,分子量为46.01 ku。对蛋白理化性质进行预测发现,SpCactus为亲水性蛋白,等电点pI为4.91。经预测,SpCactus与其他物种的IκB蛋白具有相似的功能结构域。同源性比对结果显示,SpCactus与凡纳滨对虾和中国明对虾的Cactus蛋白同源性均高达62%,相似度为73%。系统进化树分析显示,SpCactus与甲壳动物聚为一支,与无脊椎动物聚为一大支。实时荧光定量PCR(RT-PCR)检测发现,SpCactus基因在拟穴青蟹不同组织中均有表达,肌肉中的表达量最高,其次为心脏、眼柄、血液和鳃,肝胰腺中的表达量最低。LPS和金黄色葡萄球菌刺激均能显著诱导SpCactus基因的表达。本实验成功扩增了SpCactus基因全长,并进行了生物信息学分析、理化性质预测,初步探讨了其生物学功能,为进一步研究其在免疫反应中的生物学功能提供参考依据。     

Assessment of dietary lecithin and cholesterol requirements of mud crab, Scylla serrata, megalopa using semi-purified microbound diets

The effects of varying dietary lecithin and cholesterol levels on growth, development and survival of mud crab, Scylla serrata, megalopa were evaluated using six semi‐purified, microbound diets formulated to be iso‐energetic and containing three levels of supplemental lecithin (0, 20 and 40 g kg⁻¹ diet dry weight) and two levels of supplemental cholesterol (0 and 7 g kg⁻¹ diet dry weight). Fifteen megalopa were reared individually in each treatment and the nutritional value of diets was assessed on basis of mean dry weight and mean carapace width of newly settled first crab stage, as well as development time to the first crab stage and overall survival. A significant interaction between supplemental dietary lecithin and supplemental dietary cholesterol was found for final mean dry weight of newly settled crabs, and highest survival (60%) was recorded for megalopa fed diets containing the highest levels of dietary lecithin (39.7–44.1 g kg⁻¹) (diet 5 and 6) regardless of whether diets were supplemented with cholesterol; this rate of survival was identical to that of megalopa fed live Artemia nauplii. The results indicate that supplemental dietary cholesterol may not be essential for mud crab megalopa when fed diets containing sufficient levels of supplemental dietary phospholipids.     

青蟹的营养需求研究及其配合饲料研制

青蟹属（Scylla）有锯缘青蟹（Scylla serrata），拟穴青蟹（S．paramamosain），紫螯青蟹（S．tranquebarica）和榄绿青蟹（S．olivacea）等4个种。我国以拟穴青蟹占绝对优势，它有个体大、生长快、适应性强、肉味鲜美、营养丰富等特点，已成为具有重要经济价值的海水养殖蟹类之一。     

Acorn dispersal and predation patterns of four tree species by wood mice in abandoned cut-over land

We compared patterns of acorn dispersal and predation by wood mice among four tree species (Quercus serrata, Quercus crispula, Castanea crenata, and Juglans mandshurica var. sieboldiana) that are abundant in cool temperate woodlands. We devised an acorn dispersal experiment using 400 magnet-inserted acorns and a magnetic locator in a 1.8-ha study plot, which spanned a cut-over area and an adjacent deciduous forest. Ten wire mesh baskets, each containing 40 acorns (10 acorns per species), were placed on the border between these two habitat types. About 13.0% (n = 52) of the total acorns remained in the baskets, while 77.3% (n = 309) were dispersed throughout the study plot and subsequently retrieved using the magnetic locator. Microhabitat, distance, and burial depth of transported acorns were significantly different among species. In the cut-over area, J. mandshurica var. sieboldiana acorns were dispersed under fallen trees or branches and near stumps, and were buried deeply in the soil. Dispersal distances of J. mandshurica var. sieboldiana acorns were significantly greater than those of Q. serrata acorns. The number and microhabitat of transported acorns significantly differed between habitat types. J. mandshurica var. sieboldiana acorns were dispersed in the cut-over area rather than in the forest. For all four species, the numbers of acorns delivered to fallen trees or branches, stumps, and crumbled soil with overhang under any vegetation type were greater in the cut-over area than in the forest.     

饥饿对拟穴青蟹大眼幼体和Ⅰ期仔蟹蜕皮和生长的影响#br#

为了给拟穴青蟹(Scylla paramamosain)苗种培育过程中饵料的合理投喂提供基础理论数据,采用投喂-饥饿和饥饿-投喂的处理方式对拟穴青蟹大眼幼体和Ⅰ期仔蟹分别开展营养储存饱和点实验(PRS)和不可恢复点实验(PNR),研究饥饿对大眼幼体和Ⅰ期仔蟹蜕皮和生长的影响。结果显示,大眼幼体的PRS实验中,投喂时间≤2 d时,大眼幼体的蜕皮率为0;投喂时间≥4 d时,大眼幼体的蜕皮率和发育时间与连续投喂组(F组)无显著性差异;但投喂4 d-饥饿组(F4S组)大眼幼体蜕皮后Ⅰ期仔蟹的增重率和个体大小均显著小于F组(P<0.05)。大眼幼体的PNR实验中,饥饿2 d时大眼幼体的存活率仅为(30.00±13.23)%。Ⅰ期仔蟹的PRS实验中,连续饥饿组(S组)Ⅰ期仔蟹的蜕皮率为0,而投喂2 d-饥饿组(F2S组)、投喂3 d-饥饿组(F3S组)和F组3组Ⅰ期仔蟹的蜕皮率均无显著性差异。投喂1 d-饥饿组(F1S组)和F2S组Ⅰ期仔蟹蜕皮后Ⅱ期仔蟹的增重率和个体大小均显著小于F组(P<0.05),且F1S组Ⅰ期仔蟹的发育时间显著延长(P<0.05)。Ⅰ期仔蟹的PNR实验中,除饥饿1 d-投喂组(S1F组)外,F组Ⅰ期仔蟹的蜕皮率显著高于其他处理组(P<0.05);各组Ⅰ期仔蟹蜕皮后Ⅱ期仔蟹的增重率与饥饿时间呈负相关,而发育时间却与饥饿时间呈正相关;饥饿时间≥2 d时,Ⅰ期仔蟹蜕皮后Ⅱ期仔蟹的个体大小明显变小。拟合分析结果显示,大眼幼体的PRS;值是3.57 d,Ⅰ期仔蟹的PRS;和PNR;值分别是0.90 d和2.01 d。研究结果表明,新蜕皮的大眼幼体必须给予充足的饵料才可确保高的存活率,大眼幼体投喂6 d后可以适当减少投喂量,短期饥饿对Ⅰ期仔蟹蜕壳和生长的影响相对较小,Ⅰ期仔蟹的耐饥饿能力强于大眼幼体。