石鲽染色体核型分析

以石鲽(Kareius bicoloratus)成鱼为材料,采用外周血淋巴细胞培养法制备染色体标本,经Giemsa染色,拍照后进行核型分析。结果表明:石鲽的染色体核型为2n=48,48t,染色体臂数为NF=48,未发现多倍体现象,也未发现性染色体和随体。     

山东近海石鲽的同工酶分析

采用水平淀粉凝胶电泳技术研究山东半岛南岸海阳近海石鲽同工酶的组织特异性及群体遗传结构。结果表明，石鲽的眼、鳃、肌肉、心脏、肝脏、肾脏、脾脏和鳍条共8种组织的17种同工酶表达有明显的组织特异性。对肝脏和肌肉两种组织的AAT、ADH、EST、GPI、G3PDH、IDHP、LAP、LDH、MDH、MPI、PGDH、PGM、SDH和SOD等14种同工酶进行了遗传学分析，共记录了21个基因座位，其中，GPI-2^*、JDHP-2^*、MDH-1^*、MPI-1^*、PGM-3^*、SDH^*共6个基因座位呈多态，其多态座位比例为0．2857(P0.99)，平均观测杂合度和预期杂合度分别为0．0321和0．0283，平均有效等位基因数为1．3650，遗传偏离指数均大于零。石鲽和圆斑星鲽同工酶比较分析表明，二者在AAT^*、ADH^*、GPI-1^*、IDHP^*、LDH^*、MDH-1^*、MDH-2^*、MDH-3^*、MPI-1^*、PGM-1^*、PGM-2^*、SOD^*和SDH^*存在基因置换或近于基因置换，两种间的遗传距离为1．3117。     

星斑川鲽、石鲽及其杂交一代(星斑川鲽♀×石鲽♂)的线粒体DNA序列比较分析

本研究以线粒体基因细胞色素b (Cytochrome b,Cytb)和细胞色素C氧化酶Ⅰ(Cytochrome oxidase subunitⅠ,COI)为研究对象,探究比较了星斑川鲽(Platichthys stellatus)、石鲽(Kareius bicolorarus)以及杂交F1代(星斑川鲽♀×石鲽♂)的种质遗传特性.结果显示,杂交子代与亲本的碱基(A+T)含量均高于(C+G)含量,且杂交F1代与母本星斑川鲽的(C+G)相同.基于mtDNA Cytb和COI序列结果显示,石鲽与星斑川鲽遗传距离分别为0.085和0.045;杂交F1代与石鲽遗传距离分别为0.076和0.045.基于mtDNA Cytb序列显示,杂交F1代与星斑川鲽遗传距离很小,仅0.009,而两者在COI基因序列上完全一致.基于Kimura 2-parameter模型的NJ分子系统树均显示,星斑川鲽和杂交F1代聚为一支,石鲽单独聚为一支.以上结果均可得出,杂交后代在线粒体DNA上呈现明显的母系遗传.杂交后代中出现左右眼的分化,且在COI的NJ系统树中,杂交F1代与母本星斑川鲽形成的一大支又分为两支:母本星斑川鲽与杂交F1代中外观显示左眼的聚为一支,杂交F1代中外观显示右眼的单独聚为一小支.表明线粒体基因COI与杂交F1代左右眼的分化有一定关系,为进一步研究星斑川鲽♀x石鲽♂提供了参考数据.     

两种野生经济鱼类圆斑星鲽和石鲽的最新形态观测

对两种野生经济鱼类圆斑星鲽(Verasper variegatus Temminck et Schlegel)和石鲽(Kareius bicoloratus Jordan et Snyder)的外部形态、可量可比性状和可数性状进行了研究,统计分析了其体长与体质量组成、生长参数相关关系.结果表明:圆斑星鲽体长(LBy)与体高(DBy)之间的关系DBy=0.4743LBy+0.3203(r=0.9715);体长(LBy,cm)和体质量(WBy,g)之间的关系WBy=0.0158L3.0706By(r=0.9943).石鲽的体长(LBs)与体高(DBs)之间的关系DBs=0.3250LBs+2.3263(r=0.7329);体质量(WBs,g)和体长(LBs,cm)之间的关系WBs=0.25592.1238LBs(r=0.8138).这两种鱼的可量可比性状:体长/体高、体长/头长、头长/吻长和头长/眼径的比值范围比原来研究记载的更加宽泛.     

Relative contributions from exposed inshore and estuarine nursery grounds to the recruitment of stone flounder, Platichthys bicoloratus, estimated using otolith Sr:Ca ratios

In Sendai Bay, stone flounder larvae settle and spend their juvenile period in either shallow exposed inshore nursery grounds or estuarine nursery grounds. The purpose of this study is to examine the relative contributions of these two kinds of nursery grounds to the flounder population using otolith strontium:calcium ratios. Stone flounder juveniles were collected from both nursery grounds, and one- and two-year-old flounder were caught deeper in Sendai Bay. Sr and Ca content in the otoliths were measured by electron probe micro analysis. The Sr:Ca ratios in the otolith section corresponding to the early postsettlement period ranged from 3.06 to 3.85 for the exposed inshore areas with stable low temperature and high salinity conditions, and from 3.81 to 5.32 in brackish estuaries with high temperature and low salinity conditions but with large diel and tidal cyclical fluctuations. Values from an estuarine site with stable salinity ranged from 3.58 to 4.15 overlapping with both the above ranges. Rearing experiments supported our inference that the high otolith Sr:Ca ratios of juveniles inhabiting estuarine nursery grounds are attributable to higher temperature and physiological stress caused by the large diel temperature and salinity fluctuations within the estuaries. Estimation of the Sr:Ca ratio of recruited fish using the otolith section formed while in the nursery area showed that at least 20 out of 42 individuals examined originated from estuarine nursery grounds. The present study indicates that estuaries play an important role as nursery grounds for stone flounder, producing about half of the stock in spite of the small and restricted area compared with the wide expanse of the exposed inshore area.     

Predator–prey size relationship in siphon cropping between the juvenile stone flounder Platichthys bicoloratus and the bivalve Nuttallia olivacea

ABSTRACT:   The purpose of the present paper was to examine the size relationship between the juvenile stone flounder Platichthys bicoloratus and the bivalve Nuttallia olivacea , the siphon of which is important prey for the juvenile flounder. Juvenile stone flounder feed mainly on tips of the inhalant siphon of the bivalve. The maximum width of siphon tips in the stomach contents of age-0 fish could not reach that of age-1 fish, although the siphons in age-0 fish became larger as they grew. This size discrepancy indicated a limitation in the size of bivalves available to juveniles. The proportion of total cropping frequency for the siphon of the bivalve by juveniles was estimated according to the bivalve size class. Most bivalves that had cropped siphon tips ranged from 5 to 30 mm in shell length. The total cropping frequency per bivalve was particularly intense on bivalves of 10–25 mm shell length in spite of their small proportion of 24.9% of the total. This frequency intensity indicated that the size of bivalves with cropped siphons by juvenile stone flounder might depend on the ability of juveniles rather than the size composition of the bivalves.     

琼氏不动杆菌形态型Ⅰ的主要性状及系统发育分析

从以败血症感染为特征的石鲽(KareiusbicoloratusL.)中,检出了原发感染的杀鲑气单胞菌(Aeromonassalmonicida),同时还在个别病死鱼中检出了作为继发感染的另一种病原菌。经对6株纯培养菌在形态特征、理化特性等方面的鉴定,表明其为不动杆菌属(AcinetobacterBrisouand Pr啨vot1954)的琼氏不动杆菌(A.juniiBouvetandGrimont1986),但该菌表现在普通营养琼脂培养基28℃培养呈典型形态特征、在37℃培养时出现异常形态;择一个代表菌株(HQ010320B1株)送中国典型培养物保藏中心(CCTCC)予以复核鉴定与分类定名,鉴定为琼氏不动杆菌新形态型菌株并定名为琼氏不动杆菌形态型Ⅰ(AcinetobacterjuniimorphovarⅠ)。同时又择代表菌株(HQ010320B1株)进行了16SrRNA基因序列测定及系统发育分析,其基因序列长度为1427bp(在GenBank登录号为AY787213),与检索出的不动杆菌属细菌序列同源性均在97%～99%。构建了系统发育树。     

石鲽、牙鲆精子冷冻保存研究及其在人工杂交中的应用

对石鲽(Kareius bicoloratus )和牙鲆(Paralichthys olivaceus)精子冷冻保存技术进行了研究,筛选到一个优良的稀释液MPRS,用MPRS冷冻保存石鲽、牙鲆精子,冻后成活率在70%以上。在石鲽冻精和牙鲆卵的杂交实验中,杂交组受精率是 28 4%±4 55(n=3),孵化率是 42 7%±7 35(n=3),胚胎发育正常;5 000尾杂交鱼苗有102尾成功度过变态期。在牙鲆冻精和大菱鲆卵的杂交实验中,杂交组的受精率只有2 7%,部分受精卵虽能孵化出膜,但仔鱼全为畸形。这两个杂交实验,特别是石鲽冻精和牙鲆卵的杂交实验,证实了冷冻保存的精子完全可以应用在杂交育种工作中。     

石鲽基因组DNA的提取及其RAPD体系的优化

[目的]研究石鲽基因组DNA的提取及其RAPD体系的建立和优化。[方法]以石鲽为试材,按常规酚/氯仿抽提法提取其基因组的DNA,对影响RAPD反应的各因素进行优化,建立了石鲽的最佳RAPD反应体系和程序。[结果]采用常规酚/氯仿抽提法获得的DNA完全能够满足RAPD分析的要求。通过优化建立一套适合石鲽的稳定的RAPD反应体系:反应体系总体积为25μl,包括10×buff-er2.5μl,MgCl22 mmol/L,dNTPs 0.15 mmol/L,引物0.2μmol/L,模板30 ng,Taq酶1 U。扩增程序为:94℃预变性5 min,45次PCR循环(94℃变性45 s,36℃退火45 s,72℃延伸2 min和72℃延伸10 min。利用该体系对OPK和OPV系列共40条引物进行扩增,发现其中部分引物能产生稳定、清晰的条带。[结论]该体系为石鲽遗传多样性以及相关分子标记的研究奠定了基础。     

星突江鲽和石鲽正反杂交种的形态变异分析

对星突江鲽(Platichthys stellatus)、石鲽(Kareius bicoloratus)及其正反杂交种的形态特征进行了测定与分析。结果显示,正交种(星突江鲽♀×石鲽♂)和反交种(石鲽♀×星突江鲽♂)左眼型比例分别为66.9%和57.3%,介于星突江鲽(84.7%)和石鲽(0%)之间;正反杂交种体表未见似石鲽的长骨板样结构,在此大致位置处仅分布有星突江鲽体表相似的鳞片;杂交种的背鳍、臀鳍及尾鳍上均有比星突江鲽偏小、颜色偏淡的条纹,但侧线均与母本相似,正交种在胸鳍末端处弯曲,而反交种近似直线形;两杂交种可数性状的杂种指数平均值分别为20.73和20.76,可量性状的杂种指数平均值分别为27.16和26.47。卡方检验和聚类分析结果表明两杂交种的性状总体上偏向于母本;利用判别分析构建了4种鱼的判别公式,综合判别准确率达到97.50%;主成分分析显示,提取的4个主成分对总变异的累积贡献率为82.562%,主成分1对主成分3的散点图显示两杂交种在形态差异上独立于星突江鲽和石鲽。研究表明:星突江鲽与石鲽的正反杂交种在眼睛位置、鳞片和鳍的形态特征上更倾向于星突江鲽,而侧线、可数、可量性状则更多地遗传了母本的形态特征。本研究旨为星突江鲽、石鲽及其正反杂交种的形态判别和亲缘关系鉴定以及杂交选育提供理论依据。