华南沿海地区西部入海水系中间黄颡鱼的形态变异及地理分化

为探讨不同水系中间黄颡鱼 (Pelteobagrus intermedius) 的形态差异及地理分化问题,采用形态学方法, 测量12个水系中间黄颡鱼150尾样本的18个形态性状,进行主成分分析。其结果显示,前3个主成分累积贡献率为58.918%,并可将这12个水系的群体分为3簇。华南沿海地区西部诸独立入海小水系中的中间黄颡鱼群体与海南岛群体之间的相似性大于与西江干流群体的相似性。防城港地区3条直接入海的小水系的群体在形态特征上已发生了一定程度的分化,其分化主要表现在吻、眼和尾的变异。     

中间黄颡鱼群体遗传变异与亲缘生物地理

通过分析81尾采自华南西部12条水系的中间黄颡鱼(Pelteobagrus intermedius)mtDNA控制区435 bp的序列,研究其群体遗传变异及亲缘生物地理格局。结果显示,中间黄颡鱼群体间的遗传分化较小,核苷酸变异只有0.54%。12条水系的群体共有7个单倍型,其中一些现已相互隔离的水系中的群体共享同一个单倍型,提示这些水系曾经有非常密切的联系。根据嵌套进化支序分析,中间黄颡鱼可能起源于峒中河、北仑河、防城河所在的广西与越南交界地区,并通过两条途径向华南沿海西部诸独立水系和海南岛扩散,在演化过程中,曾发生片断化事件,长距离建群和持续的分布区扩张。     

中国黄颡鱼的线粒体DNA多样性及其分子系统学

基于体侧色斑、背鳍前部形态、吻长及尾柄长的差异, Ng和Kottelat(2007)将分布于中国的黄颡鱼群体划为两个物种: 北方群体为Pseudobagrus sinensis, 南方群体为P. fulvidraco。本研究通过对70个黄颡鱼标本相关形态特征的测量及对线粒体cyt b基因序列的分析, 探讨了P. sinensis物种的有效性问题。结果表明: 依据体侧色斑和背鳍前部形态的差异, 可将黄颡鱼分为对应于P. sinensis和P. fulvidraco的两种形态类型, 但对尾柄长、吻长的测量发现二者没有差异。对70条cyt b基因序列的分析结果为: 两种鱼类有1个共同的单倍型; 两种鱼类的单系性在系统发育分析中都没有得到重现, 而二者聚在一起形成获得100%支持率的单系群; 两种鱼类群体之间存在持续的基因交流(Nm = 4.7); 两种鱼类在单倍型的巢式支系分析(nested clade analysis, NCA)中没有形成各自独立的进化谱系, 所有的单倍型以不超过5步的突变全部被纳入同一个进化网络中。因此我们认为P. sinensis不是有效物种, 而应被视为黄颡鱼的一种形态类型。基于cyt b基因的序列变异, 本研究对黄颡鱼群体的遗传多样性和种群结构作了初步分析。群体的核苷酸不配对分布及Tajima’sD中性检验表明, 约在10.1-14.1万年前, 黄颡鱼在其分布范围内经历过群体扩张, 推测这可能是导致黄颡鱼群体单倍型多样度高(h = 0.857 ± 0.0014)而核苷酸多样度低( π = 0.0023 ± 0.0003)的主要原因。此外, 分析结果显示黄颡鱼群体缺乏明显的地理结构, 推测原因可能是历史上水系的连通促进了不同地理群体之间的基因交流。     

基于基因组重测序的长江上游瓦氏黄颡鱼群体遗传结构

利用基因组重测序的方法获取高通量SNP标记,分析了长江上游三峡大坝-白鹤滩大坝之间8个不同江段(太平溪、巴南、合川、岷江口、宜宾、邵女坪、桧溪、冯家坪)共136尾瓦氏黄颡鱼(Pelteobagrus vachelli)的遗传多样性和遗传分化水平,阐明了长江上游瓦氏黄颡鱼群体遗传结构。结果显示:(1)三峡库区太平溪群体和巴南群体具有较高的SNP(singlenucleotide polymorphism)数量和核苷酸多样性指数,遗传来源丰富,其遗传多样性高于其他群体;上游的岷江口、宜宾、邵女坪和冯家坪群体遗传来源单一。(2)瓦氏黄颡鱼存在3个不同的遗传分支,且不同遗传分支之间存在较大的遗传分化。(3)群体SNP数量和核苷酸多样性指数与河流坡降呈显著负相关,群体遗传分化指数与地理距离和隔离时间无显著相关性。研究结果表明,在三峡大坝-白鹤滩大坝江段,瓦氏黄颡鱼上游群体具有更低的遗传多样性,更易发生遗传漂变作用,在鱼类遗传多样性保护中需要特别关注;瓦氏黄颡鱼存在3种显著的遗传结构,应视为3个不同遗传单元进行种质资源管理。     

黄颡鱼属两种鱼类的线粒体ND4基因序列变异性分析

以东亚特有种光泽黄颡鱼(Pelteobagrus nitidus)和长须黄颡鱼(Pelteobagrus eupogon)为研究对象,采用PCR技术获得了这两种鱼类的部分线粒体DNA DN4基因及其3′端的tRNA基因碱基共约772个,用MEGA2.1软件分析了此片段序列,采用Kimura双参数模型计算遗传距离,以科属的大鳍(Hemibagrus macropterus)为外类群,用邻接法构建不同水系的光泽黄颡鱼和长须黄颡鱼的分子系统树。不同水系的光泽黄颡鱼的遗传距离在0.000—0.012之间,长须黄颡鱼的遗传距离在0.000—0.003之间,光泽黄颡鱼和长须黄颡鱼种间的遗传距离在0.099—0.108之间,从分子水平上证实了光泽黄颡鱼和长须黄颡鱼为两个有效物种。不同水系的光泽黄颡鱼遗传变异很小,除黑龙江种群外,其他水系的光泽黄颡鱼在分子系统树没有能够按水系区分开来,可能的原因为:(1)不同水系的光泽黄颡鱼之间存在频繁的基因交流;(2)东亚科鱼类的线粒体DNA的进化速率可能较小;(3)人类经济活动可能已影响到光泽黄颡鱼的种群遗传结构。     

不同地理种群东方墨头鱼(Garra orientalis)的形态分化

为确认分布于闽江、珠江、红河水系和海南岛的东方墨头鱼居群间存在的形态差异是否达到亚种或种的水平,该文将采自上述4个水系共72尾标本根据水系分为四组,比较了10个外部形态特征,并运用多变量形态度量学方法,对13个框架结构特征和23个常规特征进行分析。主成分分析结果散点图显示,海南岛居群相对集中,但仍未能与其他3个水系居群完全区分,而其他3个水系居群则完全混杂在一起。该研究结果表明,形态特征比较和形态度量学分析难以区分东方墨头鱼的不同地理居群,其种或种下分化研究需借助于分子生物学方法。     

杂交黄颡鱼与普通黄颡鱼幼鱼生长性能及耐低氧能力的比较

实验以杂交黄颡鱼(Pelteobagrus fulvidraco)和普通黄颡鱼幼鱼为实验对象, 拟通过8周的投喂生长和低氧胁迫实验, 比较研究杂交黄颡鱼与普通黄颡鱼的生长性能及耐低氧抗逆性。投喂生长实验: 经过8周的养殖, 杂交黄颡鱼平均体重为(19.60±0.88) g/尾, 显著高于普通黄颡鱼平均体重为(15.74±0.42) g/尾(P<0.05), 杂交黄颡鱼幼鱼较普通黄颡鱼幼鱼体重生长快24.52%; 杂交黄颡鱼幼鱼存活率为(87.78±1.92)%, 显著高于普通黄颡鱼幼鱼存活率(67.78±1.92)% (P<0.05), 杂交黄颡鱼幼鱼比普通黄颡鱼幼鱼存活率高 29.51%; 杂交黄颡鱼的饲料系数为1.18±0.14, 普通黄颡鱼饲料系数为1.36±0.21。低氧胁迫实验: 同时将杂交黄颡鱼和普通黄颡鱼置于在溶氧量(1.48 ± 0.27) mg/L的水体中, 分别在低氧胁迫0、6h、12h和24h后, 检测血清和肝脏中抗氧化酶活性以及脑和肝脏中缺氧诱导基因(HIF-1α)的相对表达量发现: 杂交黄颡鱼和普通黄颡鱼血清和肝脏中乳酸脱氢酶(LDH)活性、超氧化物歧化酶(SOD)活性和过氧化氢酶(CAT)活性在低氧胁迫后 6h以及总抗氧化能力(T-AOC)在低氧胁迫后 12h较低氧胁迫 0均出现显著性变化(P<0.05)且在低氧胁迫6h、12h和24h杂交黄颡鱼抗氧化酶活性均高于普通黄颡鱼; 杂交黄颡鱼和普通黄颡鱼脑和肝脏中缺氧诱导基因(HIF-1α)的相对表达量均在低氧胁迫后出现显著性上升(P<0.05)且在低氧胁迫6h、12h和 24h杂交黄颡鱼缺氧诱导基因(HIF-1α)的相对表达量均高于普通黄颡鱼。从无氧代谢能力、抗氧化能力以及缺氧诱导基因相对表达量3方面分析表明杂交黄颡鱼和黄颡鱼低氧胁迫短时间均具有一定的低氧耐受能力但随着胁迫时间延长均会出现氧化损伤且杂交黄颡鱼的耐低氧能力要显著性高于普通黄颡鱼。     

抗菌肽HBβ-C在全雄和杂交黄颡鱼对多子小瓜虫抗性差异中的作用

为研究全雄黄颡鱼(Pelteobagrus fulvidraco)、瓦式黄颡鱼(Pelteobagrus vachelli)和杂交黄颡鱼(黄颡鱼P. fulvidraco♀×瓦氏黄颡鱼P. vachelli♂)对多子小瓜虫(Ichthyophthirius multifiliis)的抗性差异, 通过生物信息学分析黄颡鱼皮肤黏液蛋白质组, 发现其血红蛋白源抗菌肽(HBβ-C)位于血红蛋白β链HBβ的碳端, 共33个氨基酸。利用化学合成的不同浓度的HBβ-C肽段进行体外抗虫实验, 研究发现其能有效杀死滋养体、包囊体和掠食体阶段的多子小瓜虫, 其中15 μg/mL的HBβ-C能在3min内杀死所有滋养体。基因表达量分析显示, 在杂交黄颡鱼的鳃和皮肤组织中, HBβ的mRNA表达量高于全雄黄颡鱼; 但在应对小瓜虫感染的过程中, 全雄黄颡鱼的HBβ mRNA转录水平快速提升, 其表达水平和上升倍率显著高于杂交黄颡鱼。蛋白表达量分析显示, HBβ在全雄黄颡鱼鳃组织中的蛋白表达量明显高于杂交黄颡鱼。免疫荧光定位结果显示, 抗菌肽HBβ-C特异地在红细胞中表达, 可以分泌并附着在滋养体上。综上所述, 相对于杂交黄颡鱼, 全雄黄颡鱼中HBβ具有更高的翻译效率, 可以更高效地应对多子小瓜虫的感染。     

雄性普通黄颡鱼与所保护受精卵间的亲缘关系分析

父母通过保护自己的子代来确保其繁殖的成功率.在普通黄颡鱼(Pelteobagrus fulvidraco)中,雄性普通黄颡鱼具有筑巢产卵保护后代的习性,其所保护的子代与其是否有亲缘关系是有待探讨的问题.本文利用10对微卫星分子标记鉴定12窝普通黄颡鱼受精卵与护卵鱼之间的亲缘关系,并对子代的遗传多样性进行分析.在单亲鉴定中,累积非父排除概率为0.9986,平均父权相对机会(RCP)在99.989%-99.999%之间,每个子代在10个微卫星位点上的累积PI值在2006.73-604464.07之间.同时在亲权鉴定分析中,发现3窝卵子的等位基因来自2个母亲,说明雄性黄颡鱼可以和2条雌性黄颡鱼发生交配;在遗传多样性分析中,黄颡鱼子代的平均等位基因数为11.7, 无偏观测杂合度值(Ho)在0.2473-0.9866之间,多态信息含量(PIC)值0.7096-0.8993之间.通过亲权鉴定分析,可以确认看护受精卵的雄性普通黄颡鱼与受精卵间的亲子关系.     

洞庭湖四种黄颡鱼基因组DNA遗传多样性的RAPD分析

以洞庭湖瓦氏黄颡鱼、光泽黄颡鱼、长须黄颡鱼、普通黄颡鱼的基因组DNA为材料对 4种黄颡鱼进行了遗传多样性随机扩增多态性DNA(RAPD)分析。通过筛选的 90个引物对 4种黄颡鱼基因组DNA的扩增 ,获得了 36个有效引物 ,并计算出了 4种黄颡鱼群体间的遗传相似系数和遗传距离 ,遗传距离最大的为普通黄颡鱼和瓦氏黄颡鱼 (D =0 9895 ) ,遗传距离最小的为普通黄颡鱼和光泽黄颡鱼 (D =0 672 0 )。同时运用聚类分析 (UPGMA)的方法建立了 4种黄颡鱼的聚类图。     

Morphological variability between wild populations and inbred stocks of a Chinese minnow, Gobiocypris rarus

Gobiocypris rarus, a small, native cyprinid fish, is currently widely used in research on fish pathology, genetics, toxicology, embryology, and physiology in China. To develop this species as a model laboratory animal, inbred strains have been successfully created. In this study, to explore a method to discriminate inbred strains and evaluate inbreeding effects, morphological variation among three wild populations and three inbred stocks of G. rarus was investigated by the multivariate analysis of eight meristic and 30 morphometric characters. Tiny intraspecific variations in meristic characters were found, but these were not effective for population distinction. Stepwise discriminant analysis and cluster analysis of conventional measures and truss network data showed considerable divergence among populations, especially between wild populations and inbred stocks. The average discriminant accuracy for all populations was 82.1% based on conventional measures and 86.4% based on truss data, whereas the discriminant accuracy for inbred strains was much higher. These results suggested that multivariate analyses of morphometric characters are an effective method for discriminating inbred strains of G. rarus. Morphological differences between wild populations and inbred strains appear to result from both genetic differences and environmental factors. Thirteen characters, extracted from stepwise discriminant analysis, played important roles in morphological differentiation. These characters were mainly measures related to body depth and head size.     

Morphological variation of the pikeperch Sander lucioperca (L.) in the southern Caspian Sea, using a truss system

A morphometric truss network system and meristic counts were used to investigate the hypothesis of population fragmentation of pikeperch Sander lucioperca among four major fishing areas in the Caspian Sea drainage. Univariate analysis of variance showed significant differences between the means of the four groups for 20 out of 32 standardized morphometric measurements (27 truss, five traditional characters) and six out of nine meristic counts. The first two canonical discriminant functions accounted for 93 and 92.3% of the total variability in morphometric and meristic characteristics, respectively. A dendogram derived from cluster analysis of Mahalanobis distances and a discriminant analysis among the groups confirmed three distinct populations in these regions, while there was a relatively high degree of overlap between all locations when using meristic characters. The most relevant morphometric characteristics were the measured distances on body heights and the caudal peduncle area. This study indicated that the morphometric truss network system was more adequate than meristic characters for separation among the populations.     

Morphometric and meristic characteristics of two groups ofLethenteron reissneri

Morphometric and meristic characteristics were compared between two genetically divergent groups ofLethenteron reissneri, collected from Hokkaido and Honshu islands, Japan. Whereas significant differences were found in some morphometric and meristic characters, the ranges overlapped somewhat in all of the characters examined between the two groups. Thus, it was difficult to distinguish exactly between the two groups using only the morphological characters. In sympatric populations of the two groups, no significant differences were found in almost all of the morphometric and meristic characters examined. The two groups ofL. reissneri, which are reproductively isolated from each other, should be regarded as sibling species. Assortative mating, based on morphological features, appears unlikely as the reproductive isolating mechanism between the two groups.     

基于形态学差异探讨秦岭细鳞鲑亚种有效性问题

秦岭细鳞鲑(Brachymystax lenok tsinlingensis Li)的亚种分类地位自命名以来一直饱受争议。研究运用单因素方差分析、主成分分析、判别分析、聚类分析及差异系数检验法, 对秦岭细鳞鲑和黑龙江流域的尖吻细鳞鲑(B. lenok lenok)和钝吻细鳞鲑(B. tumensis)的5项可数性状和34项标准化后的比例性状进行统计分析。单因素方差分析结果表明, 尖吻细鳞鲑、钝吻细鳞鲑和秦岭细鳞鲑在32项比例性状和5项可数性状上存在极显著差异(P<0.01); 主成分分析结果显示, 贡献率较大的前3个主成分的累计贡献率为92.779%; 以逐步判别分析方法选用14个判别效果较好的比例性状构建了3个细鳞鲑类群的判别函数, 综合判别率为99.4%。基于欧式距离矩阵法构建的形态学聚类图显示, 秦岭细鳞鲑和尖吻细鳞鲑距离较近, 而与钝吻细鳞鲑距离较远。据Mayr 75%亚种识别和划分规则, 34项比例性状和5项可数性状中, 尖吻细鳞鲑和秦岭细鳞鲑第一鳃弓外鳃耙数目的差异系数大于1.28, 而钝吻细鳞鲑和秦岭细鳞鲑有31项比例性状和2项可数性状的差异系数高于1.28。据分析结果, 结合秦岭细鳞鲑地理隔离的事实及其与尖吻细鳞鲑和钝吻细鳞鲑的形态差异, 推断秦岭细鳞鲑与黑龙江流域内的尖吻细鳞鲑的形态差异程度至少已达亚种水平。     

Geographical differentiation of Aphanius dispar (Teleostei: Cyprinodontidae) from Southern Iran

The Arabian cyprinodontid Aphanius dispar (Rüppell, 1829) is known to show considerable morphological variation. It has remained unknown, however, whether this variation is a result of environmental differences or allopatric divergence owing to geographical isolation. In this study, 11 populations of A. dispar from three geographically separated basins were analysed, that is, the Makran Basin (I, one river system), the Hormuzgan Basin (II, five rivers and three hot springs) and the Helleh Basin (III, two hot springs) in southern Iran. Statistical analyses do not indicate significant differences between the fishes from river and hot spring habitats (T‐test, p < 0.05), which is also supported by the Canonical Discriminant Analysis (CDA). Nevertheless, morphometric and meristic characters of the fishes, as well as otolith morphology and morphometry, demonstrate that six phenotypic characters discriminate the A. dispar populations of the three basins, that is, (1) predorsal distance (Prdd.SL), (2) head length (HL.SL), (3) pelvic fin length (Lplf.SL), (4) number of pelvic fin rays, as well as relative length of both the (5) medial part and (6) rostrum of the otolith. In addition, these characters display a consistent pattern of variation, thus providing support for the assumption that the phenotypically different A. dispar populations are a result of geographical isolation and not related to environmental differences. It is likely that the geological history of the drainage systems caused isolation event(s) that may date back to the Pleistocene (1.8 million years before present). The high phenotypic differences might suggest that the A. dispar populations from the three studied basins represent separate subspecies or even species.     

Geographic variation in Austrolebias luteoflamulatus Vaz‐Ferreira, Sierra & Scaglia (Cyprinodontiformes, Rivulidae)

The morphological variation in the annual killifish Austrolebias luteoflamulatus was studied using meristic and pigmentation characters analysed with univariate and multivariate analysis, and morphometric characters analysed with a geometric morphometry approach (thin plate spline). The results showed that morphological variation in this species was associated with a geographical pattern. In the case of meristic and pigmentation characters, the strongest distinction occured in males, while females were more similar among different drainages. Males from Rocha (eastern Uruguay) exhibited the greatest differences from the others. The number of dorsal fin rays and bars on the flanks were the characters that contributed most to this segregation. When morphometrics were considered, the three populations analysed could be distinguished well from each other in both sexes. Some of the characters that contributed to this pattern were dorsal fin origin, position of eyes and length of snout. Since pigmentation characteristics and some characters of the dorsal fin in males were involved in the differences observed, sexual selection could have played an important role in the differentiation of populations of this species. Since Austrolebias luteoflamulatus has a restricted geographical distribution, conservation of this species should include consideration of these populations individually since they might represent different evolutionary units.     

Population dynamics of fisheries stock enhancement

A comparative analysis was performed on sixteen morphometric characters, in three different juvenile sterlet Acipenser ruthenus L. populations. Specimens were collected from a wild population in the Serbian part of the Danube River ( n  = 46), from aquaculture stocks in the Czech Republic originating from Russia ( n  = 40), and aquaculture stocks in the Slovak Republic originating from Slovakian part of the Danube River ( n  = 28). Average values for total length were 29·9 ± 3·9 cm, 29·1 ± 3·7 cm and 27·3 ± 7·7 cm for Serbia, Czech Republic and Slovak Republic, respectively. Populations were compared using t ‐test and sequential Bonferroni correction for multiple comparisons was applied in order to determine significant differences between them. Results of analysis showed that all three populations differed in prebarbel length, interocular distance and maximum head width. Although all these characters are head‐related, head length itself was very uniform among all populations. The natural population from the Serbian part of the Danube River differed from the populations reared in aquaculture in seven morphometric characters. The two populations reared in aquaculture consistently showed lower morphological variability than the wild population, even though they had different genetic backgrounds (Russia and Slovakia). Future genetic studies will show if this tendency is caused by a reduction in genetic variability.     

Morphometric and meristic diversity of the species Scolopendra cingulata Latreille, 1829 (Chilopoda: Scolopendridae) in the Mediterranean region

Scolopendra cingulata Latreille, 1829 is the commonest scolopendromorph species in the Mediterranean area. However, its morphological and geographic variation has not been examined so far, and therefore robust hypotheses about the factors that have shaped that variability are lacking. We examined, using multivariate methods for 19 morphometric and meristic characters, the morphological variation of that species using a comprehensive sample of 503 adult specimens from 130 restricted geographic localities. The localities were distributed in three major geographic areas (Balkans, Asia Minor-Middle East, and Italy) in order to discern biological entities and to estimate the morphological relationships between populations and geographic regions. Results showed significant differences between the three geographic groups. Characters such as the distance between the paramedian sulci of the 7th and 8th tergite, the number of antennal segments, the number of spines on the dorsal side of 21st prefemur and number of spines on the 21st prefemoral process significantly discriminated populations of S. cingulata along a west-east geographic gradient. Both eastern (Balkan) and easternmost populations (Asia Minor-Middle East) showed higher mean values of antennal articles and spines of 21st prefemur than the Italian populations. No significant morphological variation was discovered between the sexes of S. cingulata apart from cephalic width and distance between the two paramedian sulci of the 7th and 8th tergite. Based on certain morphometric differences among the three major geographic regions, we suggest that S. cingulata originated in the east and colonized south European area from two different geographic routes. It is suggested that west-east morphological trend of S. cingulata is related either to the prominent palaeogeographic events of the area or to the competition with S. oraniensis Lucas, 1846 in the western Mediterranean.