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Construction of AFLP-based genetic linkage maps for the chinese shrimp Fenneropaeneus chinensis

chinese science Bulletin 2008年第8期53卷 1205-1216页

作者： tian yi1,2,3,4, kong jie1↑ & wang weiji1 1 yellow sea fisheries research institute, chinese academy of fishery science, qingdao 266071, china 2 College of Life science and Technology, Dalian fisheries University, Dalian 116023, china 3 institute of Oceanology, chinese academy of sciences, qingdao 266071, china 4 Graduate University of chinese academy of sciences, Beijing 100049, china chinese Acad fisheries Sci Yellow Sea Fisheries Res Inst Qingdao 266071 Peoples R China. op.uky.edu

Fenneropaeneus chinensis is an important species in marine fishery resources and aquaculture in china. A genetic linkage map is essential for improving the efficiency of its breeding by marker-as- sisted selection and identifying commercially important genes. Linkage maps of F. chinensis were constructed with an F2 mapping population (110 progenies) using amplified fragment length polymor- phic (AFLP) marker in this study. Fifty-five AFLP primer combinations produced 532 AFLP markers fitting for map strategy in mapping family. The markers with 3:1 segregating ratios were analyzed using F2 intercross model for the common linkage map, while the markers with 1:1 ratio were analyzed using the pseudo-testcross strategy. The maps of male, female and common were constructed. The female map included 103 markers that formed 28 linkage groups, covering a total length of 1090 cM. All mark- ers were linked with the linkage groups. Segregation distortion was observed for 6 of 103 markers in the female map. The average distance between markers was 14.53 cM and ranged from 4.4 to 24.8 cM. The male map included 144 markers that formed 35 linkage groups. Ten markers remained unlinked in male map. Segregation distortion was observed for 7 of 144 markers in the male map. The total dis- tance of male map covered 1617 cM. The average distance between markers was 16.36 cM. The male map was 32.6% longer than the female map, which may reflect sex-specific recombination rates in chinese shrimp. The common map was composed of 216 markers, including in 44 linkage groups covering a total distance of 1772.1 cM. Two markers remained unlinked. No distorted markers of 216 markers were shown in the common map. The distance between markers was 10.42 cM. An average estimated genome size for the chinese shrimp was 2420 cM, which was consistent with the relative size of the Penaeid genome. The distribution of AFLP markers was relatively even in chromosomes of Chi- nese shrimp maps. The linkage analysis

关键词： AFLP 海洋渔业虾中国

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Estimation of genetic parameters and breeding values in shrimp F enneropenaeus chinensis using the REML/BLUP procedure

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Acta Oceanologica Sinica 2011年第1期30卷 78-86页

作者： ZHANG tianshi kong jie LUAN Sheng wang Qingyin LUO Kun tian yia College of Life sciences and Technology Ocean University of China Qingdao 266003 China Key Laboratory for Sustainable Utilization of Marine fisheries Resources Ministry ofAgriculture Yellow Sea Fisheries Research Institute Chinese Academy of Fishery SciencesQingdao 266071 China institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China

An analysis of a selection experiment was used to assess the impact of various animal model struc- tures on REML estimates of variance components. The analyses were carried out based on 162 d body mass （BM） of 1 287 animals from 21 paternal half-sib groups of Fenneropenaeus chinensis. Estimated breeding values （EBV） of BM of all individuals were estimated using eight statistical models （A, AB, ABC, ABDC, ABMFC, ABMDC, ABFDC and ABMFDC） and BLUP （best linear unbiased prediction）. These models were designed involving factors such as sex, spawn date as fixed effects, maternal genetic effects, full-sib family effects as random effects, mean BM of families at tagging and age at recording （covariate）. The results demonstrate the importance of correct interpretation of effects in the data set, particularly those that can influence resemblance between relatives. The data structure and the particular model that was applied markedly influenced the magnitude of variance component estimates. Models based on few effects obtained upward biased estimates of additive genetic variance. The accuracy of genetic parameters and breeding value es- timated by ABFDC model was higher than other models. The results imply that additive genetic direct value, full-sib family effects, and covariance effects besides sex and spawn date as fixed effects were very important for estimating genetic parameters and breeding value of body mass. This model had a heritability estimate of 162 d BM of 0.44. The comparison of the efficiency of selection based on breeding values or phenotypic value revealed great difference： average breeding value of the best 24 families selected by the 162 d BM breeding value and phenotype were 0.577 g and 0.366 g, respectively, representing a 36.57% higher efficiency in the former. In conclusion, selection based on breeding value was more effective than selection based on phenotypic value. Our results indicate that effects influencing the magnitude of estimates should be tak

关键词： Fenneropenaeus chinensis animal model BLUP breeding value

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Estimates of genetic and phenotypic parameters for weight and length in Paralichthys olivaceus(Temminck et Schlegel)

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Acta Oceanologica Sinica 2011年第6期30卷 58-64页

作者： tian Yongsheng XU tianjun LIANG You CHEN Songlin Key Laboratory for Sustainable Utilization of Marine fisheries Resources Ministry of Agriculture Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences Qingdao 266071 China Laboratory for Marine Living Resources and Molecular Engineering College of Marine ScienceZhejiang Ocean University Zhoushan 316000 China

The complete diallel cross design was employed to estimate the genetic parameters of the growth of Japanese flounder （Paralichthys olivaceus）. A total of 60 full-sib families were cultivated and two growth-related traits, body weight （BW） and body length （BL）, were examined at average 2, 3 and 8 months of age respectively, with 1 800 individuals measured in each age group （30 per family）. Based on the additive-dominance-maternal-effect genetic analysis model, the restricted maximum likelihood approach was used to estimate various （co）variance components. The results showed that narrow-sense heritability estimates of BW and BL were respectively： 0.29 ± 0.01 and 0.22 ± 0.02 at 2 months of age, 0.32 ± 0.02 and 0.30 ± 0.04 at 3 months of age, 0.48 ± 0.04 and 0.40 ± 0.05 at 8 months of age; broad-sense heritability estimates were respectively： 0.44 ± 0.02 and 0.54 ± 0.04 at 2 months of age, 0.35 ± 0.01 and 0.36 ± 0.03 at 3 months of age, 0.49 ± 0.03 and 0.45 ± 0.04. All heritabilities were statistically significant （P 0.05）. Additive genetic correlations between BW and BL at 2, 3 and 8 months of age were consistently positive and highly significant （P 0.01）： 0.93 ± 0.02, 0.95 ± 0.03 and 0.92 ± 0.03 respectively. Maternal effect was significant （P 0.05） only at 2 months of age, and was not detected at 3 and 8 months of age. According to the heritability estimates, the mass selection strategy should be efficient for the breeding of Japanese flounder.

关键词： Paralichthys olivaceus heritability genetic correlation genetic mating design diallel cross maternal effect

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Programmed cell death in Laminaria japonica(Phaeophyta) tissues infected with alginic acid decomposing bacterium

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Progress in Natural science:Materials International 2004年第12期 32-36+105页

作者： wang Gaoge 1*, LIN Wei 1, ZHANG Lijing 2, YAN Xiaojun 1 and DUAN Delin 1**(1. institute of Oceanology, chinese academy of sciences, qingdao 266071, china 2. yellow sea fisheries research institute, chinese academy of fishery sciences, qingdao 266071, china) instituteofOceanology ChineseAcademyofSciencesQingdao266071China yellowseafisheriesresearchinstitute ChineseAcademyofFisherySciencesQingdao266071China

TdT mediated dUTP biotin nick end labeling (TUNEL) is a sensitive and valid method for detecting DNA cleavage in programmed cell death (PCD). Using this method, DNA cleavage was observed in Laminaria japonica sporophytic tissues, which were infected with alginic acid decomposing bacterium. It was found that DNA cleavage occurred 5 min after the infection, the fragments with 3′ OH groups of cleaved nuclear DNA increased with time of infection and spread from the infection site. Although no typical DNA ladder (200 bp/180 bp) was detected by routine agarose gel electrophoresis, the cleavage of nuclear DNA fragments of 97～48.5 kb could be detected by pulsed field gel electrophoresis (PFGE). By using CaspGLOW TM fluorescein active caspase 3 staining method, caspase 3 activity has been detected in response to the infection of alginic acid decomposing bacterium. Our results are similar to the observations in hypersensitive response (HR) of higher plant, suggesting that the rapid cell death of L. japonica infected by alginic acid decomposing bacterium might be involved in PCD, and indicating that the occurrence of PCD is an active defense process against the pathogen's infection.

关键词： alginic acid decomposing bacterium caspase-3 Laminaria japonica hypersensitive reaction (HR) programmed cell death (PCD) TUNEL.

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C-banding pattern and nucleolar organizer regions of Cynoglossus semilaevis Günther, 1873

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Progress in Natural science:Materials International 2006年第7期16卷 769-772页

作者： WU Di, ZHANG Shicui, ZHUANG Zhimeng, PANG Qiuxiang, wang Changliu and WAN Ruijing ( Department of Marine Biology, Ocean University of china, qingdao 266003, china yellow sea fisheries research institute, chinese academy of fishery sciences, qingdao 266071, china Department of Biology, Yantai Normal University, Yantai 264025, china) Ocean Univ china Dept Marine Biol Qingdao 266003 Peoples R China chinese Acad fishery Sci Yellow Sea Fisheries Res Inst Qingdao 266071 Peoples R China Yantai Normal Univ Dept Biol Yantai 264025 Peoples R China

The C-banding pattern and nucleolar organizer regions of the metaphase chromosomes of Cynoglossus semilaevis are reported. The interstitial regions in all chromosomes including the pair of sex chromosomes had positive C-bands, and the 6th and 12th pairs of chromosomes were entirely stained in most cases. Some chromosomes such as the 1st, 2nd, 7th, 8th, 9th and 10th pairs showed C-bands at centromeric or distal ends. The C-banding heretochromatin occupies 30.03% of the total chromosome surface in C. semilaevis, which is similar to that of amphibians such as Mixophyes fasciolatus (30. 2% ) and M. schevilli (20. 7% ), but is rather lower than that of cephalochordate Branchiostoma belcheri (54. 3 % ). Silver staining revealed a single pair of nucleolar organizer regions ( NORs) located in the telomeric regions of Chromosome 2. The association of NORs with heterochromatin observed in vertebrates also occurs in C. semilaevis as the telomeric regions of Chromosome 2 are always stained positively with C-banding.

关键词： Cynoglossus semilaevis karyotype C-banding NORs.

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3株水产动物病原弧菌主要分泌性蛋白的鉴定及其分泌性序列的分析

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水产学报 2011年第11期35卷 1701-1710页

作者：刘笋王秀华黄倢中国水产科学研究院黄海水产研究所农业部海洋渔业资源可持续利用重点开放实验室上海海洋大学水产与生命学院

从3株水产动物病原菌鳗弧菌MN、鳗弧菌3101、费氏弧菌培养液中分别获取了分泌性蛋白,进行SDS-PAGE,对表达量较高的5条蛋白区带进行MALDI-TOF/TOF质谱鉴定表明,它们分别是鳗弧菌MN金属蛋白酶（empA-MN）、鳗弧菌3101金属蛋白酶（empA-310... 详细信息

从3株水产动物病原菌鳗弧菌MN、鳗弧菌3101、费氏弧菌培养液中分别获取了分泌性蛋白,进行SDS-PAGE,对表达量较高的5条蛋白区带进行MALDI-TOF/TOF质谱鉴定表明,它们分别是鳗弧菌MN金属蛋白酶（empA-MN）、鳗弧菌3101金属蛋白酶（empA-3101）、锌金属蛋白酶（zinc empA）、费氏弧菌VFMJ111094蛋白（VFMJ111094）和费氏弧菌ES114的外膜蛋白（OMP）。根据所鉴定的序列设计引物,分别从鳗弧菌MN、鳗弧菌3101和费氏弧菌中扩增出相应基因,克隆后测定emp-MN、empA-3101、zinc empA、VFMJ11-1094和OMP相应基因的序列,经在线软件SignaIP 3.0分析,确定emp-MN、empA-3101、zincempA、VFMJ11-1094和OMP均存在不同的分泌性信号肽序列angMN-35、ang3101-35、ang3101-25、vf-38和vf-23,通过在线软件PSORT分析表明,所有信号肽均定位在细胞的周质空间或细胞外膜上。

关键词：水产动物病原菌分泌性蛋白 MALDI-TOF/TOF质谱分析分泌性序列细胞定位

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中国对虾杂交优势对自然感染白斑综合征病毒的抗病力分析

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水产学报 2007年第1期31卷 68-75页

作者：李素红张天时孟宪红孔杰中国水产科学研究院黄海水产研究所中国水产科学研究院黄海水产研究所山东青岛266071 上海水产大学生命科学与技术学院上海200090

一个包括42个全同胞家系的中国对虾选育群体,荧光标记后每个家系各取40尾健康个体混养,自然发病并确认感染白斑综合征病毒(white spot-syndrome virus,W SSV)。发病1周后将所有个体取出,统计各家系的存活率,并对活体利用巢式聚合酶链式... 详细信息

一个包括42个全同胞家系的中国对虾选育群体,荧光标记后每个家系各取40尾健康个体混养,自然发病并确认感染白斑综合征病毒(white spot-syndrome virus,W SSV)。发病1周后将所有个体取出,统计各家系的存活率,并对活体利用巢式聚合酶链式反应(polymerasechain reaction,PCR)进行W SSV检测。对存活率进行分析,结果显示:4个家系的存活率在60%以上,7个家系的存活率在20%以下;存活率与第一次扩增阳性率、第二次扩增阳性率呈不显著负相关。第一次扩增阳性率与第二次扩增阳性率呈极显著正相关。结合两次扩增的阳性率分析得出:青岛野生群体内自交产生的一个家系和韩国群体养殖家系与乳山野生群体杂交产生的一个家系在抗病力上显著优于其它家系;经独立样本t检验表明,群体间交配组合的存活率差异不显著,因此家系选育可能是进行抗W SSV品种选育的重要途径。多重比较结果发现:韩国群体养殖家系(♂)与乳山野生群体(♀)杂交后代的存活率(56.20%±8.75%)显著高于韩国群体养殖家系自交后代(22.50%±2.50%)(P26.95)与其反交后代(3.85)差异显著。

关键词：中国对虾巢式PCR 白斑综合征病毒

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用微卫星DNA技术对中国对虾人工选育群体遗传多样性的研究

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水产学报 2005年第1期29卷 6-12页

作者：张天时刘萍李健孔杰王清印中国水产科学研究院黄海水产研究所中国水产科学研究院黄海水产研究所山东青岛　266071中国海洋大学海洋生命学院山东青岛　266003 山东青岛　266071

利用微卫星技术对中国对虾人工选育群体第1代和第6代群体的遗传多样性进行了分析。对10个微卫星位点进行了扩增,共产生74个等位基因,每个位点产生的等位基因数从3到13不等。在两个群体中,所观察到的等位基因数都比有效等位基因数多。多... 详细信息

利用微卫星技术对中国对虾人工选育群体第1代和第6代群体的遗传多样性进行了分析。对10个微卫星位点进行了扩增,共产生74个等位基因,每个位点产生的等位基因数从3到13不等。在两个群体中,所观察到的等位基因数都比有效等位基因数多。多态信息含量PIC值0.5567～0.8877,说明这10个微卫星位点在中国对虾中具有较高的信息含量。两个群体的平均杂合度分别为0.6400(CP1)、0.6300(CP6),并通过计算基因型的P值,确定了对Hardy Weinberg平衡的偏离情况。对Fis值的计算表明两个群体内共有5个微卫星位点存在杂合度观察值过剩的现象。两个群体的Shannon多样性指数分别为1.6830、1.7382,整个选育群体(两个群体作为一个群体)的遗传多样性指数为1.7742。从遗传多样性所占的比例来看,96.415%的遗传变异是来自群体内,只有3.585%的遗传变异是来自群体间。两个群体间的相似性系数高达0.9187,彼此间的遗传距离仅为0.0848,体现出人工选育群体的遗传分化程度较低。结果均说明第6代群体还有较大的选育潜力,可以继续保持遗传效应,最终保证选种育种工作的成功。

关键词：中国对虾微卫星遗传变异杂合度

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大菱鲆肠道蛋白酶的分离纯化及性质的初步研究

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水产学报 2005年第5期29卷 42-47页

作者：王海英薛长湖孙谧王跃军王清印中国水产科学研究院黄海水产研究所中国海洋大学生命科学与技术学部中国水产科学研究院黄海水产研究所山东青岛266071 山东青岛266003

以大菱鲆肠道为材料,对大菱鲆消化生理中起重要作用的肠蛋白酶的分离纯化条件和理化性质进行了研究。经Tris-HCl缓冲液抽提,硫酸铵分级分离,DEAE-Sepharose Fast Flow离子交换层析、Sephadex G-100凝胶过滤层析分离纯化,获得大菱鲆肠蛋... 详细信息

以大菱鲆肠道为材料,对大菱鲆消化生理中起重要作用的肠蛋白酶的分离纯化条件和理化性质进行了研究。经Tris-HCl缓冲液抽提,硫酸铵分级分离,DEAE-Sepharose Fast Flow离子交换层析、Sephadex G-100凝胶过滤层析分离纯化,获得大菱鲆肠蛋白酶Ⅰ的电泳纯制品,并对该酶的性质进行了研究。结果显示:纯酶的分子量约为58kD。纯酶最适反应pH为9.0,最适反应温度50℃;pH稳定范围6.0~11.0,30℃以下酶活性稳定;Mn2+可激活大菱鲆肠蛋白酶Ⅰ,Cu2+、Zn2+、Ag+、Fe3+则对酶活性有抑制作用;巯基蛋白酶抑制剂显著抑制大菱鲆肠蛋白酶Ⅰ的活性,金属蛋白酶抑制剂对大菱鲆肠蛋白酶Ⅰ无影响。50℃、pH9.0条件下以双倒数作图法得大菱鲆肠蛋白酶Ⅰ催化酪蛋白水解的Km值为2.92g.L-1。

关键词：大菱鲆肠蛋白酶纯化性质

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黄姑鱼群体遗传多样性的AFLP分析

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水产学报 2006年第5期30卷 640-646页

作者：韩志强高天翔王志勇庄志猛苏天凤中国海洋大学教育部海水养殖重点实验室集美大学水产学院中国水产科学研究院黄海水产研究所中国水产科学研究院水产种质资源与养殖技术重点开放实验室山东青岛266003 山东青岛266071 福建厦门361021 广东广州510300

对青岛和厦门黄姑鱼群体的遗传多样性进行了AFLP分析,5对选择性引物在两个群体47个个体中,共扩增出461个位点,多态位点265个。青岛和厦门群体的多态位点比例、Nei遗传多样性指数和Shannon遗传多样性指数分别为51.70%、51.99%,0.1022、0.... 详细信息

对青岛和厦门黄姑鱼群体的遗传多样性进行了AFLP分析,5对选择性引物在两个群体47个个体中,共扩增出461个位点,多态位点265个。青岛和厦门群体的多态位点比例、Nei遗传多样性指数和Shannon遗传多样性指数分别为51.70%、51.99%,0.1022、0.0996,0.1643、0.1622;两个群体遗传多样性在同一水平上。基因分化系数Gst、Shannon遗传多样性指数和AMOVA分析均显示黄姑鱼的遗传变异主要来源于群体内个体间,而群体间无明显的遗传分化。群体的显性基因型频率分布和位点差异数分布显示两个群体有基本相同的群体遗传结构。结果表明,黄姑鱼青岛和厦门群体间无明显的遗传差异,群体间有明显的基因交流。

关键词：黄姑鱼 AFLP 遗传多样性

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