大菱鲆胃质子泵(H~+/K~+ ATPase)与消化机能发生的关系     

迟良  徐世宏  肖志忠  马道远  林帆  赵春彦  肖永双  武宁宁  刘清华  李军 《海洋与湖沼》2014,45(3):644-650

胃质子泵(H+/K+ATPase)是胃酸分泌的关键酶。本试验采用RACE和PCR方法从大菱鲆的胃组织中提取RNA克隆得到了H+/K+ATPaseα亚基cDNA全长序列。结果表明:大菱鲆H+/K+ATPaseα亚基序列全长3467 bp,开放阅读框为2964 bp,编码988个氨基酸。与GenBank上其它物种比对发现,大菱鲆H+/K+ATPaseα亚基与斑鳜同源性最高,为89%。进化树分析发现,H+/K+ATPase在进化上具有物种特异性。经RT-PCR和荧光定量PCR检测,大菱鲆H+/K+ATPase在胚胎孵化后22d开始表达,晚于大菱鲆胃腺出现的时间(16 d),说明大菱鲆胃腺的发育完成并不代表胃功能的完善。另外,大菱鲆H+/K+ATPase除了在胃中大量表达之外,在食道中的表达量也很高。结合组织学观察,作者认为,大菱鲆H+/K+ATPase在食道中大量表达是因为在发育上食道是胃的前体,因此保留了分泌H+/K+ATPase的能力。同时通过整体原位杂交试验表明:大菱鲆H+/K+ATPase会首先在食道的末端和胃的贲门处表达。本研究为进一步了解海水鱼类的消化机制提供了理论基础。  相似文献   

大菱鲆(Scophthalmus maximus)微卫星序列的开发与分析     

李猛  马爱军  岳亮  邹杰  王广宁  田岳强  马本贺  夏丹丹 《海洋与湖沼》2013,44(5):1365-1371

采用FIASCO (Fast Isolation by AFLP of Sequences Containing repeats)法和标记初步筛选法, 进行了大菱鲆(Scophthalmus maximus)微卫星标记开发的研究, 分析了大菱鲆基因组中微卫星DNA序列的特征, 并进行了拟作图标记的初步筛选。结果表明, FIASCO法的富集效率为78.56%, 二次筛选出655个阳性克隆, 测序后经分析有469条含有微卫星位点的单一序列, 共得到597个微卫星位点。其中完美型占51.76%, 非完美型占34.67%, 复合型占13.57%; 核心序列的重复次数在3—96次之间, 平均重复数为13.39次; 重复单元类别最多的是(CA/GT)n, 占77.6%。利用Primer Premier 5.0共设计413对引物, 其中有360对能够得到稳定表达的产物; 再经标记初筛试验得知, 183对引物(50.8%)的PCR产物有多态性, 其中110个微卫星位点(30.6%)符合遗传连锁图谱作图标准。  相似文献   

大菱鲆育苗期的细菌病研究   总被引：8，自引：0，他引：8  

姚志刚  丁天宝 《海洋科学》2004,28(9):10-12

检测出大菱鲆(Scophthlmus maxims)幼体发育过程中3种病原菌,即仔鱼期病原菌两种：链球菌(Streptococcus equisimilis)和溶藻弧菌(Vibrio alyinolyticus),稚鱼期一种病原菌即副溶血弧菌(Vibio parahaemolyticus),并进行药物敏感试验,利用药物青霉素,呋喃唑酮、四环素进行有效治疗,使育苗成活率达到24％,高于目前国外平均水平。  相似文献   

Ciliate Uronema marinum is the causative agent of scuticociliatosis in farm raised turbot Scophthalmus maximus     

DU Guangxun  QU Lingyun  SHANG Kun  SUN Chengjun  WANG Chen  GAO Ping 《海洋湖沼学报（英文）》2019,(5):1726-1735

To identify the pathogen that causes scuticociliatosis in farmed-raised turbot Scophthalmus maximus , we isolated a ciliate from the brain tissue of an infected turbot and identifi ed it as Uronema marinum based on morphological and molecular evidence. We then infected the turbots in artifi cial laboratory settings with pure cultured U . marinum . The infected turbots showed syndromes similar to those observed in naturally infected ones. Furthermore, microscopic examination and PCR detection confi rmed the presence of the ciliate in the tissues of those laboratory-infected turbots. To our best knowledge, this is the fi rst report of scuticociliatosis caused by U . marinum in farm-raised turbot S . maximus in China.  相似文献   

Estimation of genetic parameters for upper thermal tolerances and growth-related traits in turbot Scophthalmus maximus     

WANG Xin′an  MA Aijun  HUANG Zhihui  SUN Zhibin  CUI Wenxiao  QU Jiangbo  YU Hong 《海洋湖沼学报（英文）》2019,(5):1736-1745

An abnormally high temperature produces a stress response in turbot causing large economic losses in the turbot aquaculture industry of China. A genetic improvement of the upper thermal tolerance (UTT) of turbot could allow cultured fi sh to adapt. A genetic evaluation of UTT is required for determining the practicability of including this trait into a breeding program. In this study, data were recorded from a temperature tolerance test conducted on 3 200 individual turbots from 32 full-sib groups. A cross-sectional linear model and a cross-sectional threshold probit model were used to analyze the test-period survival and a cross-sectional threshold logit model was used to analyze the test-day survival. In addition, phenotypic and genetic correlations between body weight and survival data were estimated. The estimated heritability values obtained from the cross-sectional linear model (CSL), the cross-sectional threshold (probit) model (THRp), and the cross-sectional threshold (logit) model (THRl) were 0.247 9±0.108 3, 0.288 3±0.161 2, and 0.106 9±0.045 2, respectively. The correlation coeffi cients among the full-sib family estimated breeding values (EBVs) obtained from the three models were greater than 0.998 6 and all models produced an almost identical family ranking. The accuracies of selection obtained with the CSL, THRp, and THRl model were 0.773 8, 0.775 4, and 0.784 4, respectively, the greatest from the THRl model. The genetic correlations between body weight and survival data EBVs from the CSL, THRp, and THRl models were 0.020 1,-6.201 1×10^-4 , and -3.115 4×10^-4 , respectively, and the phenotypic correlations between the two traits were -0.837 1 and -0.667 1, respectively. The findings of this study provide background information to determine the best strategy of selection for the genetic improvement of UTT in turbot.  相似文献   

免疫增强剂紫锥菊(Echinacea purpurea)提取物对大菱鲆(Scophthalmus maximus)头肾内非特异性免疫基因表达量的影响     

秦志华  潘柳婷  董文宾  单虎 《海洋与湖沼》2018,49(2):340-345

通过给大菱鲆(Scophthalmus maximus)注射不同浓度的紫锥菊(Echinacea purpurea)提取物,测定其非特异性免疫指标的变化水平,研究其对大菱鲆的头肾内非特异性免疫分子基因相对表达量的影响,进而为其在大菱鲆养殖生产过程中推广应用提供科学依据。试验选取体重(250±20)g的大菱鲆作为试验动物,分别注射10、20、40mg/m L浓度的紫锥菊提取物,试验共进行28d,注射试验结束后,分别从高、中、低剂量组及空白对照组选出6尾大菱鲆,分别取出大菱鲆的头肾并提取总RNA。采用?Ct法进行目标基因的荧光定量分析,再应用SPSS17.0软件对获得的实验数据进行单因素方差分析。研究结果表明,紫锥菊提取物能够不同程度地提高大菱鲆头肾内非特异性免疫分子基因相对表达量。注射紫锥菊提取物28d后对大菱鲆头肾中Lysozyme基因相对表达量的影响显著(P0.05),对C3补体基因相对表达量的影响极显著(P0.01),对Transferrin基因相对表达量的影响显著(P0.05),对TGF-β1基因相对表达量的影响极显著(P0.01),对IL-1h基因相对表达量的影响极显著(P0.01),本试验研究表明,紫锥菊提取物能够显著提高大菱鲆头肾内非特异性免疫分子基因的相对表达量。  相似文献   

大菱鲆(Scophthalmus maximus)蛋白质二硫键异构酶SmPDIA3的表达分析和功能验证     

唐启政  孙志宾  王新安  马爱军  杨双双  黄智慧 《海洋与湖沼》2019,50(2):409-419

本研究利用SMART-RACE技术克隆了大菱鲆SmPDIA3基因。SmPDIA3基因cDNA序列全长2083bp,包括1479bp的开放阅读框,编码492个氨基酸,GenBank登录号：MG765516。组织表达分析发现：SmPDIA3基因在肠、鳃、肝脏等组织中均有表达,其中在肝脏中表达量最高,脑中最低。进一步研究了温度胁迫后SmPDIA3基因在肠和肝脏组织中的表达变化规律,发现随着胁迫温度的升高,SmPDIA3基因的相对表达量总体上升,并在28℃时达到最大。利用原核表达技术,构建了pET-28a-PDIA3原核表达载体,IPTG诱导后融合蛋白主要在上清中表达。将上清中的蛋白分离纯化后,利用Western blot技术验证了目的蛋白的准确性。将纯化后的目的蛋白浓缩后,利用BCA蛋白浓度测定试剂盒测得蛋白浓度为243.18μg/mL。最后设计变性溶菌酶的复性实验,验证了SmPDIA3蛋白具有分子伴侣功能,能够辅助变性蛋白的正确折叠。  相似文献   

急性热应激对大菱鲆血液生化指标的影响   总被引：1，自引：0，他引：1  

孟振  张鸿丽  刘新富  贾玉东  刘滨  曲江波 《海洋科学》2020,44(1):122-131

为探讨大菱鲆(Scophthalmusmaximus)大规格幼鱼热耐受性和热应激过程中敏感指标的变化,本研究设定18℃、21℃、24℃和27℃4个温度梯度,采用1℃/h的升温速率,对驯化温度为18±1℃的苗种进行96 h急性胁迫实验,统计各温度组96 h成活率和测定血浆部分生化指标的变化情况。结果表明,在pH 7.85、盐度29.5和溶解氧高于6.5 mg/L的环境条件下,平均体质量183.65±15.99 g的大菱鲆幼鱼96h高起始致死温度(UILT50)为28.05℃;温度和胁迫时间对血浆肾上腺素(EPI)、皮质醇(Cortisol)、血糖(GLU)、还原型谷胱甘肽(GSH)、超氧化物歧化酶(SOD)和碱性磷酸酶(AKP)含量/活性都存在显著影响,除GSH外,温度和胁迫时间存在显著的交互影响作用;热应激组血浆EPI含量随胁迫时间延长呈逐渐上升趋势,同一胁迫时间以21℃组含量最高;热应激组Cortisol含量的响应时间随温度升高相应提前、随胁迫时间呈先升后降的趋势,均在48 h达最高值; 24℃和27℃组GLU含量随胁迫时间呈先升后降趋势,胁迫3h～6h即达最高值,上升幅度与温度正相关,21℃组则呈逐渐升高的趋势;热应激组GSH含量和SOD活性仅在胁迫初期(0 h～3 h)与对照组存在显著差异;热应激组AKP活性处理12 h显著低于对照组, 48 h后则显著高于对照组,且升高幅度与温度呈正相关。  相似文献   

Annual and spatial variation in the abundance length and condition of juvenile turbot (Psetta maxima L.) on nursery grounds on the west coast of Ireland: 2000–2007     

Paula S. Haynes  Deirdre Brophy  David McGrath  Roisin O'Callaghan  Stephen Comerford  Paul Casburn 《Journal of Sea Research》2010,64(4):494-504

Turbot (Psetta maxima Linnaeus) is a high value commercially exploited marine flatfish which occurs in European waters, from the Northeast Atlantic to the Arctic Circle, the Baltic and Mediterranean Sea. In Ireland, turbot are the most valuable commercial non-quota species. Very little is known about their population dynamics in the wild, in particular during the sandy beach nursery phase of the life history. In 2000, a survey was established to assess flatfish species on nursery grounds on the west coast of Ireland. Eleven sandy beaches were assessed for 0+ turbot by beach seining, over an eight year period (2000–2007) during the months of August and September. The objective of the study was to estimate juvenile turbot abundance and size structure to determine if any spatial and annual trends existed. Large scale variability in the recruitment of fish to nursery grounds may be indicative of fluctuations in the adult stock. Turbot were found to recruit to five beaches consistently over the eight year period. Temporal and spatial variability in the relative abundance and length of turbot was discerned, with no apparent overall trend. However, certain nursery grounds were shown in most of the years examined to support higher abundances of turbot in comparison to other areas over the eight year period. Turbot abundances on nursery grounds were significantly correlated with mean spring sea temperatures during the pelagic stage. The condition of turbot did not significantly differ on an annual or spatial scale. Mean densities of 0+ turbot along the Irish coast were found to be similar and at times higher than other areas in Europe, ranging from 0.1 (± 0.3) individuals 1000 m^− 2 to 18.5 (± 6.9) individuals 1000 m^− 2. Mean turbot total length on beaches ranged from 3.8 cm (± 0.6) to 6.6 cm (± 4.3). The observed spatial and temporal variability in abundance and length highlights the need for long-term studies when assessing juvenile flatfish populations. Results from the present study have provided much needed baseline data on wild juvenile turbot populations which is severely lacking for this species both on an Irish and on a European scale.  相似文献   

大菱鲆(Scophthalmus maximus)选育家系的构建和培育技术研究   总被引：5，自引：2，他引：3  

马爱军  王新安  薛宝贵  黄智慧  杨志  曲江波 《海洋与湖沼》2010,41(3):301-306

采用巢式设计方法和人工采卵授精技术,对大菱鲆大规模家系的构建与培育技术进行研究。按照1雄配2雌的原则,选取英国、法国、丹麦和挪威4个群体的大菱鲆进行定向交配,并对大菱鲆早期发育阶段苗种进行了环境标准化和一级、二级、三级数量标准化培育。结果表明,构建父系半同胞家系的初始成功率不高,1/3父系半同胞家系的初始构建成功率≤20%;由于家系成功初始构建的不同步性,拉长了家系的培育时间,对同一发育阶段家系早期培育的同步性较差,导致对处于同一发育阶段、但在不同时期培育的家系所进行的环境标准化效果不好,每次数量标准化时,存在部分全同胞家系间数量差异显著。基于早期阶段家系构建和培育存在的问题,提出了拟解决的方法和对策,为大规模建立大菱鲆家系提供参考。  相似文献