中国水域真海豚头骨特征的初步研究

测定了 8个真海豚 (Delphinussp .)头骨的 3 5个形态学指标 ,并与其它水域真海豚的相应数据比较。结果表明 ,在喙的相对长度和绝对长度、颅基长、喙端至外鼻孔和内鼻孔的长度、上齿列长以及下颌骨长等方面 ,中国水域的真海豚与东北太平洋水域的长喙真海豚 (D .capensis)非常接近。中国水域真海豚的喙长与颧宽之比为 1 68～ 2 0 3 ,平均 1 85± 0 1 0 ;喙长与喙宽之比为 3 5 6～ 4 0 3 ,平均 3 79± 0 1 7,也完全符合长喙真海豚的鉴别特征。因此 ,中国水域的真海豚在分类上应属于长喙真海豚。     

用mtDNA序列鉴定一头小布氏鲸标本

测定了采自浙江省瑞安市的一头须鲸类标本的线粒体DNA（mitochondrial DNA,mtDNA)细胞色素b(cytochrome b,cyt b)基因369bp和控制区（control region)933bp的序列,通过与已发表的须鲸类同源序列比对,发现与西太平洋和日本水域的布氏鲸的cyt b基因和控制区分别有6.78%-7.05%和13.30%－14．40％的序列差异,而与来自所罗门群岛的布氏鲸之间cyt b基因的序列完全相同,控制区的序列也仅相差一个碱基（0.28%)。提示与邻近的西太平洋和日本海的普通布氏鲸在遗传上有显著区别,而可能与所罗门群岛的布氏鲸为同一种,即小布氏鲸（Balaenoptera edeni)。同时表明,应用分子生物学手段为进行鲸肉及其制品的种类鉴定是可行和有效的。     

加州Santa Monica海湾鳍足类的生态学

研究了加州Santa Monica海湾鳍足类的生态学.从1997-2007年乘船调查了277次,发现海狮(Zalophus californianus)是最常见的动物(89%,见到的次数为1393次),其次是港海豹(Phoca vitulina richardsi,8%,n=131)和北象海豹(Mirounga angustirostris,1%,n=15).在29%的遇见次数(观察到瓶海豚205次)中,发现海狮(偶尔也发现港海豹)与瓶鼻海豚集群(Tursiops truncatus);短喙真海豚(Delphinus delphis)与长喙真海豚(D.capensis)在53% 的遇见次数(遇见真海豚次数n=155)中,发现短喙真海豚(Delphinus delphis)与长喙真海豚(D.capensis)集群;一般在沿岸水域(离岸边距离<500 m)见到海狮和港海豹,但在整个海湾也能见到,表现出这两个物种对海底峡谷的偏爱.北象海豹仅见于近海,主要在海底峡谷附近. 经常看到海狮、港海豹和北象海豹游动(50%,n=728)、进行热调节(14%,n=205)、以及取食(3.2%,n=47),但几乎见不到有社会性活动(0.21%,n=3).     

长爪沙鼠线粒体细胞色素b 基因的遗传变异及地理分化

本文基于6 个种群52 个个体的线粒体细胞色素b (Cyt b)基因全序列的遗传变异分析,探讨了我国长爪沙鼠种群的遗传结构和种群历史。在1140 bp Cyt b基因的碱基序列中,共发现了74个变异位点,其中转换和颠换位点分别为65和9个,共定义了37个单倍型。种群差异分析表明包头种群与其他种群之间都存在着显著的遗传差异,而其他种群间遗传差异则不显著。52 个个体在系统发生树中明显聚为两支(Clade A 和Clade B),其中Clade B 仅含有贺兰山东部种群中的3 种单倍型,而其余的单倍型分布于Clade A 中。来自Clade A 的49个个体的歧点分布分析结果呈单峰状,提示长爪沙鼠种群可能在历史上经历了种群增长和扩张事件,这一结果同时也得到了Tajima 检验结果(D=-1.86,P<0.05)和Fu 检验结果(Fs=-21.89,P <0.05) 的支持。通过种群扩张系数(τ)和分子钟的推算提示该种群扩张事件大约发生于11 万年前。我们的研究结果表明,长爪沙鼠在第四纪冰期中形成了明显的隔离分化,而贺兰山东部可能作为冰期避难所,包含于所有的谱系分支中,在末次间冰期种群进行了强烈的扩张,从而形成当今的分布格局。     

中国水域瓶鼻海豚的mtDNA控制区序列变异性分析

测定了30头中国水域瓶鼻海豚（Tursiops sp.)mtDNA控制区5′端424bp的序列,结合已发表的中国水域其它瓶鼻海豚的mtDNA控制区序列,共发现54个变异位点,定义了37种单元型。中国水域瓶鼻海豚的两个形态型之间没有共享单元型,且具有8个鉴别位点。基于最大似然法和邻接法的系统发生分析均把单元型聚类为分别代表两个形态型的支系。形态型之间的核苷酸歧异度为5.58%,超过了其它海豚类种间的序列歧异水平,支持把这两个形态型划分为两个独立的种,即T.truncatus和T.aduncus的观点。虽然两种瓶鼻海豚的分布区在台湾海峡一带出现重叠,但相互之间缺乏基因流动,提示两者可能已出现了显著的生殖隔离。     

绿翅短脚鹎线粒体基因组测序分析

本文对绿翅短脚鹎Ixos mcclellandii线粒体基因组进行了测序分析。结果显示,绿翅短脚鹎线粒体基因组序列全长17 838 bp(Gen Bank登录号:KX640824),具有2个开放阅读框重叠区,即ATP6～ATP8(10 bp)和ND4L～ND4(7 bp),还有一些重叠发生在蛋白质基因和其相邻的tRNA基因之间。除COXⅠ、ND3的起始密码子分别为GTG、ATA外,其余11个蛋白质基因的起始密码子均为ATG。9个蛋白质基因以TAA、AGA或AGG为终止密码子,其余则以T(COXⅢ和ND4)或TA(ND2和ND4L)为终止密码子。蛋白质基因使用频率最高的密码子是CUA(217次)和ACC(205次),而GGU和GCG的使用频率最低,均为24次。tRNA基因分布在rRNA基因和蛋白质基因之间,长度为64～75 bp。通过分析已报道的雀形目Passeriformes鸟类线粒体控制区结构,发现了3种不同类型的控制区结构:1)仅存在1个控制区; 2)有2个长度相近且序列高度相似的控制区; 3)有2个高度异质的控制区。绿翅短脚鹎线粒体基因组含有2个高度相似的控制区(相似度91. 6%),长度分别为1 116 bp和1 144 bp,二者仅在控制区开始和末端部位的序列有所不同。     

三种海豚线粒体COI基因的序列分析

对瓶鼻海豚(Tursiops truncatus)、中华白海豚(Sousa chinensis)和糙齿海豚(Steno bredanensis)的线粒体DNA COI基因进行了测序分析。PCR产物约700bp。扩增产物直接测序,去除引物序列后分别获得643、618和618bp的核苷酸序列。碱基组成平均为,T:31·07%,C:26·13%,A:27·27%,G:15·50%,GC含量为41·63%,其中碱基G的含量明显较低。与Gen Bank中9种鲸的同源序列比对,去除部分端部序列后得到597个比对位点,包括141个简约信息位点,43个单突变子,无插入/缺失位点。12种鲸的种间序列差异较大,其序列变异度在2·1%～17·1%之间。597个比对位点编码199个氨基酸,其中有9个氨基酸发生改变,其中一个氨基酸突变可将齿鲸亚目和须鲸亚目分开。NJ系统树表明,海豚科形成单系类群,瓶鼻海豚和中华白海豚的亲缘关系较近。上述分析表明,COI基因可用于鲸类的种类鉴定和系统发育分析。     

利用mtDNA序列及形态特征重新鉴定福建长乐搁浅喙鲸

<正>中国大陆沿岸属大陆棚水域,常见的齿鲸物种以近岸型的海豚科(Delphinidae)、小抹香鲸科(Kogiidae)及鼠海豚科(Phocaenidae)动物为主(王丕烈,2012),而喙鲸科(Ziphiidae)发现纪录相对较少,近年来仅由王丕烈等(2011)针对喙鲸科的中喙鲸属(Mesoplodon)标本进行检视校正,认为中国大陆沿岸存在柏氏中喙鲸     

麦穗鱼线粒体基因组序列测定及分析

利用麦穗鱼Pseudorasbora parva和相关鱼类的部分线粒体基因序列,设计出2对长批引物和30对短批引物,采用基于长PCR的2次PCR扩增法测定并注释麦穗鱼线粒体基因组全序列。结果表明,麦穗鱼线粒体基因组长16600bp,A+T含量为58.9%,37个基因位置及组成与其它硬骨鱼一致,均由13个蛋白编码基因、22个tRNA、2个rRNA基因和1个控制区(D-loop)组成。其中L链仅含8个tRNA(Pro、T yr、Ser、Ala、Asn、Cys、Glu、Gln)及ND6基因,其余基因皆由H链编码。基因排列紧密,间隔序列共计13处64bp,长度从1～32bp不等;基因重叠区7处23bp,重叠碱基数在1～7bp之间。13个蛋白编码基因中,除COI起始密码子为GTG外,其余均以ATG为起始密码子;有8个基因(ND1、ND2、COI、ATP6、ATP8、ND4L、ND5、ND6)3’端有完全的TAA或TAG终止密码子,其它5个基因终止密码子为不完整的TA(ND3和ND4)或T(COⅡ,COⅢ,Cyt b)。除tRNASer(AGY)外,其余21个tRNA基因的二级结构均为典型的三叶草结构。预测的lrRNA二级结构共有6个结构域,53个茎环结构,srRNA二级结构包含43个茎环结构。控制区(D-loop)存在3个结构区:终止序列区(TAS)、中央保守区(CSB-F、CSB-D)和保守序列区(CSB-1、CSB-2、CSB-3),其中TAS与DNA复制终止相关,出现茎环结构。     

苏云金芽胞杆菌cyt2Ba16基因的克隆表达

[目的]获得苏云金芽胞杆菌QL32-1中杀虫晶体蛋白基因,并分析其杀虫活性。[方法]采用cyt基因通用引物、两步法染色体步移的方法对QL32-1中cyt基因进行鉴定和全长克隆,再构建重组质粒pET-28a-2Ba16,并转化到大肠杆菌BL21(DE3)中,经IPTG诱导表达,最后对表达蛋白进行杀虫活性分析。[结果]克隆得到1个全长783 bp cyt2B基因,其编码260个氨基酸,推导分子量约29.69 kDa,此氨基酸序列与已知的Cyt2Ba1蛋白同源性最高,为92.4%。该基因被国际杀虫晶体蛋白基因命名委员会正式命名为cyt2Ba16。杀虫活性分析表明:cyt2Ba16对双翅目的库蚊(Culex quinquefasciatus)具有显著的杀虫活性,LC50为9.73μg/mL。[结论]cyt2Ba16基因是一个对库蚊具有较高杀虫活性的新型cyt基因,为QL32-1菌株的应用提供理论基础。     

Phylogeography and alpha taxonomy of the common dolphin (Delphinus sp.)

The resolution of taxonomic classifications for delphinid cetaceans has been problematic, especially for species in the genera Delphinus, Tursiops and Stenella. The frequent lack of correspondence between morphological and genetic differentiation in these species raises questions about the mechanisms responsible for their evolution. In this study we focus on the genus Delphinus, and use molecular markers to address questions about speciation and the evolution of population structure. Delphinus species have a worldwide distribution and show a high degree of morphological variation. Two distinct morphotypes, long-beaked and short-beaked, have been considered different species named D. capensis and D. delphis, respectively. However, genetic differentiation between these two forms has only been demonstrated in the Pacific. We analysed samples from eight different geographical regions, including two morphologically defined long-beaked form populations, and compared these with the eastern North Pacific populations. We found high differentiation among the populations described as long-beaked instead of the expected monophyly, suggesting that these populations may have evolved from independent events converging on the same morphotype. We observed low genetic differentiation among the short-beaked populations across a large geographical scale. We interpret these phylogeographical patterns in the context of life history and population structure in related species.     

TAXONOMIC STATUS AND GEOGRAPHICAL CRANIAL VARIATION OF COMMON DOLPHINS (DELPHINUS) IN THE EASTERN NORTH ATLANTIC

The common dolphin has a widespread distribution and is relatively abundant in the temperate to subtropical waters of the eastern North Atlantic. However, it is not known whether different species, subspecies, or populations occur in this region. We examined 393 common dolphin skulls obtained from both stranded and bycaught individuals collected between 1901 and 2005. The series included skulls of 152 females and 199 males, from animals ranging in body length from 93 to 230 cm and 105 to 244 cm, respectively. The ranges of total body length, skull size, RL/ZGW ratio and maximum upper alveolar (tooth) count of common dolphins in the eastern North Atlantic overlapped with those of both short- ( D. delphis ) and long-beaked ( D. capensis ) species found off the Californian coast. However, in the absence of additional data, the common dolphin in the eastern North Atlantic is regarded here as a large form of Delphinus delphis . Sexual dimorphism and possible sex-linked characters were identified within the sample. Results of the current study indicate some population differentiation within the eastern North Atlantic, with common dolphins off Portugal showing segregation in morphometric characteristics from common dolphins in other areas.     

Genetic differentiation among recently diverged delphinid taxa determined using AFLP markers

In the mid-1990s, a new common dolphin species (Delphinus capensis) was defined in the northeast Pacific using morphological characters and mitochondrial DNA (mtDNA) sequences. This species is sympatric with a second species, Delphinus delphis; morphological differences between the two are slight and it is clear they are closely related. Does the phenotypic distinction result from only a few important genes or from large differences between their nuclear genomes? We used amplified fragment length polymorphism (AFLP) markers to broadly survey the nuclear genomes of these two species to examine the levels of nuclear divergence and genetic diversity between them. Furthermore, to create an evolutionary context in which to compare the level of interspecific divergence found between the two Delphinus taxa, we also examined two distinct morphotypes of the bottlenose dolphin (Tursiops truncatus). A nonmetric multidimensional scaling analysis clearly differentiated both Delphinus species, indicating that significant nuclear genetic differentiation has arisen between the species despite their morphological similarity. However, the AFLP data indicated that the two T. truncatus morphotypes exhibit greater divergence than D. capensis and D. delphis, suggesting that they too should be considered different species.     

Bycatch and strandings programs as ecological indicators for data-limited cetaceans

An integrated approach of using strandings and bycatch data may provide an indicator of long-term trends for data-limited cetaceans. Strandings programs can give a faithful representation of the species composition of cetacean assemblages, while standardised bycatch rates can provide a measure of relative abundance. Comparing the two datasets may also facilitate managing impacts by understanding which species, sex or sizes are the most vulnerable to interactions with fisheries gear. Here we apply this approach to two long-term datasets in East Australia, bycatch in the Queensland Shark Control Program (QSCP, 1992–2012) and strandings in the Queensland Marine Wildlife Strandings and Mortality Program (StrandNet, 1996–2012). Short-beaked common dolphins, Delphinus delphis, were markedly more frequent in bycatch than in the strandings dataset, suggesting that they are more prone to being incidentally caught than other cetacean species in the region. The reverse was true for humpback whales, Megaptera novaeangliae, bottlenose dolphins, Tursiops spp.; and species predominantly found in offshore waters. QSCP bycatch was strongly skewed towards females for short-beaked common dolphins, and towards smaller sizes for Australian humpback dolphins, Sousa sahulensis. Overall, both datasets demonstrated similar seasonality and a similar long-term increase from 1996 until 2008. Analysis on a species-by-species basis was then used to explore potential explanations for long-term trends, which ranged from a recovering stock (humpback whales) to a shift in habitat use (short-beaked common dolphins).     

The use of carcasses for the analysis of cetacean population genetic structure: a comparative study in two dolphin species

Advances in molecular techniques have enabled the study of genetic diversity and population structure in many different contexts. Studies that assess the genetic structure of cetacean populations often use biopsy samples from free-ranging individuals and tissue samples from stranded animals or individuals that became entangled in fishery or aquaculture equipment. This leads to the question of how representative the location of a stranded or entangled animal is with respect to its natural range, and whether similar results would be obtained when comparing carcass samples with samples from free-ranging individuals in studies of population structure. Here we use tissue samples from carcasses of dolphins that stranded or died as a result of bycatch in South Australia to investigate spatial population structure in two species: coastal bottlenose (Tursiops sp.) and short-beaked common dolphins (Delphinus delphis). We compare these results with those previously obtained from biopsy sampled free-ranging dolphins in the same area to test whether carcass samples yield similar patterns of genetic variability and population structure. Data from dolphin carcasses were gathered using seven microsatellite markers and a fragment of the mitochondrial DNA control region. Analyses based on carcass samples alone failed to detect genetic structure in Tursiops sp., a species previously shown to exhibit restricted dispersal and moderate genetic differentiation across a small spatial scale in this region. However, genetic structure was correctly inferred in D. delphis, a species previously shown to have reduced genetic structure over a similar geographic area. We propose that in the absence of corroborating data, and when population structure is assessed over relatively small spatial scales, the sole use of carcasses may lead to an underestimate of genetic differentiation. This can lead to a failure in identifying management units for conservation. Therefore, this risk should be carefully assessed when planning population genetic studies of cetaceans.     

Genetic and family structure in a group of 165 common bottlenose dolphins caught off the Japanese coast

The biological and genetic structure of common bottlenose dolphins (Tursiops truncatus) that migrate seasonally near Japan remains largely unknown. We investigated the genetic and family structure in a group of 165 common bottlenose dolphins caught off the coast of Japan using mitochondrial DNA (mtDNA) and 20 microsatellite DNA markers. Phylogenetic analysis of the mtDNA control region sequences suggested that the dolphins were related more closely to oceanic types from Chinese waters than other geographic regions. The information on sex, sexual maturation and age together with the genetic markers revealed a strong likelihood for 37 familial relationships related mostly to maternity and an under‐representation of juvenile female offspring. The maternal dolphins had a similar offspring‐birth interval as the coastal types from North Atlantic Ocean, but a slightly younger first‐progeny age. The sex bias in the captured group was particularly marked towards an over‐representation of males among the young and immature dolphins, whereas the mature adults had an equal number of males and females. These results should be useful for future comparative biological, genetic and evolutionary investigations of bottlenose dolphins from the North Pacific Ocean with those from other regions.     

Population differentiation in the Pacific white-sided dolphin Lagenorhynchus obliquidens inferred from mitochondrial DNA and microsatellite analyses

We investigated genetic diversity and differentiation of the Pacific white-sided dolphin (Lagenorhynchus obliquidens) in Japanese coastal waters and offshore North Pacific by analyzing mitochondrial DNA and nuclear microsatellite variation. A total of 519 bp of the mitochondrial control region was sequenced and five microsatellite locus were genotyped for 59 individuals. A high level of haplotypic diversity (h=96.1%), moderate level of nucleotide diversity (pi=1.65%) and average expected heterozygosity (HE=0.66-0.76) were within an extent of those reported for other odontocetes. Consistent genetic difference between the samples from Japanese coastal Pacific-Sea of Japan and offshore North Pacific was indicated by analyses of molecular variance (AMOVAs) based on mtDNA and microsatellite variations, comparison of genetic variabilities, and geographical distributions of mtDNA haplotypes and microsatellite alleles. This result suggests that Pacific white-sided dolphins in each of the above two areas belong to different populations between which gene flow has been severely restricted. The low genetic diversity and mtDNA genealogy of the population in Japanese coastal waters suggest that it originated from a small population that colonized the Sea of Japan or that experienced population reduction when this Sea was isolated from the North Pacific during a glacial period in the Late Pleistocene.