用mtDNA控制区序列鉴定长须鲸标本

测定了采自长江口和杭州湾交汇海域的一头死亡大型须鲸骨骼标本的线粒体DNA(mt DNA)控制区序列(Control region)976 bp(登录号MF781125)、细胞色素C氧化酶Ⅰ基因(COⅠ)序列642 bp(登录号MG010134)和Cyt b序列307 bp(登录号MG010133)。通过与Gen Bank已发表的同源序列blast结果表明,与美国加利福尼亚海域长须鲸(Balaenoptera physalus)的控制区序列相似度达99%,仅在756 bp处有一个碱基T和C转换的差异;基于HKG+G模型,使用UPGMA聚类分析法和最大似然法(ML)构建的系统发育树与blast结果一致,故将标本鉴定为长须鲸,推测该个体来源于北太平洋。获取的细胞色素C氧化酶Ⅰ和Cyt b序列存在多个终止密码子,无法获取同源性较高的序列,推断可能为线粒体假基因。     

用mtDNA序列鉴定出上海海域 须鲸科物种大村鲸

测定了采自上海海域的一头死亡须鲸骨骼样本的线粒体DNA（mtDNA）细胞色素b基因（Cyt b）序列330 bp（登录号MK295815）、细胞色素C氧化酶Ⅰ基因（COⅠ）序列206 bp（登录号MK317953）和控制区（control region）序列231 bp（登录号MK317954）。通过GenBank中的BLAST分析,结果表明,样本的上述三部分序列与大村鲸（Balaenoptera omurai）对应序列的相似性均达到100%,基于最大似然法（ML）法构建的系统发育树与BLAST结果一致,故将标本鉴定为大村鲸,为上海海域首次记录。     

北部湾涠洲岛海域栖息布氏鲸种群的物种鉴定

布氏鲸(Bryde’s whale)是广泛分布于温带和热带海域的一类中等体型须鲸,通常认为存在小布氏鲸(Balaenoptera edeni edeni)和布氏鲸(B. e. brydei)两个亚种。然而,有研究表明它们应该被划分为两个独立物种,即近岸小型布氏鲸(B. edeni)和远洋大型布氏鲸(B. brydei)。由于两者外部形态极其相似,并且存在同域分布现象,很难基于外观进行准确的物种鉴定。近年来,广西北部湾涠洲岛海域出现一个稳定的布氏鲸栖息种群,但目前尚不清楚属于哪种布氏鲸。研究采集了涠洲岛布氏鲸种群中两个体的粪便样品,从其中一份样品成功提取基因组DNA,并基于线粒体Cyt b和COΙ基因序列开展物种鉴定和遗传分析,鉴定结果为小布氏鲸。此外,还鉴定出涠洲岛海域同年死亡的一头须鲸也为小布氏鲸。据此推测涠洲岛水域栖息的布氏鲸种群可能是小布氏鲸。研究首次基于粪便样品,采用分子生物学技术,成功开展了活体布氏鲸的物种鉴定。这种基于非损伤采样的物种鉴定方法值得进一步优化并推广应用。     

北京地区7种常见嗜尸性蝇类的COI基因序列分析及DNA条形码的建立

嗜尸性蝇类在命案死亡时间和现场推断方面有着十分重要的应用, 而DNA 条形编码技术能摆脱对虫卵和幼虫的饲养以及后续物种鉴定方面专业知识的依赖, 有助于实现现场采集蝇类样本的快速鉴定。本研究采集了北京地区7个嗜尸性蝇类优势种共77个个体的样本, 测定了所有个体线粒体DNA 上细胞色素C氧化酶亚基Ⅰ（COI）基因1 120 bp的序列。基于序列的系统发生分析显示, 同一物种不同个体的序列均以高达99%的支持值聚集在一起。序列间的分歧统计表明这些蝇类在物种内的个体分歧不超过1%, 而不同物种间的净分歧均超过7.74%, 最高可达14.85%。滑动窗口分析表明, 在整个序列区段种间差异位点存在较平均的分布。通过测定COI基因的序列, 建立了北京地区7个嗜尸性蝇类优势种的DNA条形码, 据此实现了对这些物种准确、快速、简单的区分和鉴定, 同时也为后续应用于物种鉴定的种属特异性位点之筛选提供了基础数据。     

基于线粒体COI基因序列的壮真蝎与普洱真蝎的分子鉴定

对蝎类物种的传统分类主要依靠形态和行为特征,但由于该类群种间形态特征极为相似,物种的划分和鉴定困难。为弥补传统分类方法的不足,本研究以线粒体细胞色素氧化酶亚基Ⅰ(COI)基因作为分子标记,对形态相似的壮真蝎Euscorpiops validus和普洱真蝎E.puerensis进行分子水平的物种鉴定。采用PCR扩增测序获得壮真蝎与普洱真蝎共24个样本的COI基因部分片段序列(660 bp),进行了遗传距离、系统发育及单倍型网络图分析。结果显示:壮真蝎15个样本中共检测到4个单倍型,单倍型之间的相似度为99.3%~99.8%;普洱真蝎9个样本中共检测到4个单倍型,单倍型之间的相似度为99.6%~99.8%;2种蝎的种间序列相似度为90.1%~90.6%,单倍型间的稳定差异核苷酸位点数为61个。壮真蝎与普洱真蝎种内平均遗传距离分别为0.004 0、0.002 3,种间平均遗传距离为0.103 9,且种间遗传距离为种内的34.6倍。此外,分子系统发育树显示壮真蝎与普洱真蝎的单倍型序列各自聚为2个单系枝,且具有很高的分枝自举值(100%)。单倍型网络图结果也显示壮真蝎与普洱真蝎8个单倍型明显分为2大类群,且壮真蝎的单倍型HAP2与普洱真蝎的单倍型HAP7之间的突变步数高达62步。上述结果不仅进一步确认壮真蝎与普洱真蝎为2个不同的物种,且表明线粒体COI基因可用于开展真蝎属Euscorpiops物种的分子鉴定。     

在长江口发现的长鬚鲸

1965年12月2日,在长江口宝山和崇阳县之间的横沙岛因搁浅捕获到一头活鲸,经鉴定为长须鲸的Balac-nopteraphysalus Linnaeus,该鲸为雌性幼体。据文献记载长须鲸分布在我国的黄海北部、台湾海峡和南海。在长江口内发现长须鲸还是第一次。横沙岛是位于长     

基于ITS2和psbA-trnH序列的金钗石斛DNA条形码鉴定研究

石斛药材品种繁多,外观鉴定较困难,为保证药材临床疗效,药材品种溯源是关键。本研究以60份贵州赤水产金钗石斛(包括鱼肚兰,竹叶兰和圆茎兰三种种质,各20份)为研究对象,提取其基因组DNA并作为PCR扩增模板,扩增核糖体ITS2和叶绿体psb A-trn H基因片段,并进行测序和相关分析。结果显示,金钗石斛三种种质核糖体ITS2共有序列相似度达96.69%,两两比较,相似度均为100.0%,长度分别为圆茎兰491 bp、竹叶兰492 bp、鱼肚兰493 pb;3种种质叶绿体psb A-trn H共有序列相似度达87.32%,两两比较,相似度均达99.0%以上,长度分别为圆茎兰465 bp、竹叶兰534 bp、鱼肚兰469 bp。两条基因在种内遗传变异较小,均不能区分金钗石斛三种种质。在中药材DNA条形码鉴定系统中,ITS2可将金钗石斛与其他物种区分开,但psb A-trn H不能,可将二者组合作为贵州赤水产金钗石斛的DNA条形码。     

基于SNP分子标记的凹叶木兰遗传多样性初步研究

凹叶木兰是我国特有的木兰属植物,仅分布于我国四川和云南两省相邻地区,目前已被列入中国物种红皮书名录,属易危物种。该研究以采自四川南部麻咪泽和美姑大风顶两个自然保护区的野生凹叶木兰为材料,利用以PCR和测序为基础的SNP分子标记方法,初步研究了凹叶木兰的遗传多样性。结果表明:在扩增出的4条510bp长的序列上,平均在73bp左右的序列长度上能够检测到一个SNP位点,说明两个自然保护区内不同居群的凹叶木兰具有较高遗传多样性;序列间相似度达97%,与引物所对原序列相似度达36%,表明该序列适宜进行凹叶木兰遗传多样性研究。研究结果为进一步开展凹叶木兰遗传多样性的研究及其保护政策的制定提供了参考。     

基于线粒体基因组探讨鲸偶蹄类七种动物的系统发育关系

利用作者已测定的赤麂(Muntiacus muntjak)线粒体全基因组序列和从GenBank检索到的鲸偶蹄类有代表性的长须鲸、河马、奶牛、绵羊、猪、羊驼6种动物的线粒体全基因组序列,按顺序分别连接各自的13个蛋白编码基因、13个氨基酸序列、2个rRNA基因和22个tRNA基因成一个氨基酸序列或核苷酸序列,用DNASTAR软件统计碱基长度和组成;分析奶牛、绵羊和赤麂两两之间蛋白编码基因的序列差异;用MEGA计算7种动物12S和：16S rRNA基因的遗传距离;基于连接在一起的13个蛋白编码基因的氨基酸序列,用NJ法构建系统关系树。结果显示：①偶蹄目反刍亚目牛科的绵羊和同亚目鹿科的赤麂先聚为一亚支,然后与该亚目牛科的奶牛并为一支;猪形亚目猪科的猪和胼足亚目驼科的羊驼并为一支;鲸目须鲸亚目须鲸科的长须鲸和偶蹄目猪形亚目河马科的河马并为一支。②赤麂与绵羊的亲缘关系更近。③推测赤麂与绵羊和奶牛的分歧时间分别约在14．7和16．0百万年前。     

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。     

北部湾涠洲岛水域发现上颌和须板异常的布氏鲸:对保护的启示

2018年初以来,北部湾涠洲岛附近出现了布氏鲸（Balaenoptera edeni）的活动。一头上颌与须板异常的小布氏鲸个体引发热议,其视频在网络上广泛流传。我们在船只调查时,目击该个体10次,其以单独活动为主（90%）,主要出现在涠洲岛到斜阳岛之间的水域,最小凸多边形家域面积为14km2,核心家域面积为166.9km²。然而,在 2019年3月30日我们发现该个体已死亡漂浮在海面,根据尸体腐烂状况来推测,该个体的死亡时间大约为3~5日,死亡原因不明。根据照片和现场解剖分析,推测该小布氏鲸的上颌和鲸须异常可能是被渔网或绳索缠绕导致的。由于无法从外形上确认属于哪一个亚种,因此我们测定了该个体的线粒体DNA D-loop（mitochondrial DNA, mtDNA）和细胞色素b（cytochrome b, Cyt b）基因,分别得到909bp和395bp的序列,经比对和系统发育重建,发现该个体属于近岸分布的小布氏鲸亚种（Eden’s whale, B. e. edeni）。由于小布氏鲸具有一定季节迁移特性,我们无法判断造成其上颌伤害的渔网或绳索是否在中国水域。尽管如此,仍然建议当地部门应加强宣传,减少渔网等海洋垃圾的丢弃和排放,为小布氏鲸营造一个安全的栖息环境。     

Species identification and likely catch time period of whale bones from South Georgia

Skeletal remains of baleen whales killed during the onset of 20th century commercial whaling lie scattered across the shores and abandoned whaling stations of the subantarctic island of South Georgia. Here we report on genetic species identification of whale bones collected from South Georgia using standard historical DNA protocols. We amplified and sequenced short fragments of the mitochondrial DNA (mtDNA) control region from 281 available bone samples. Of these, 231 provided mtDNA sequences of sufficient quality and length (174–194 bp) for species identification: 158 bones were identified as humpback whale (Megaptera novaeangliae), 51 bones were identified as fin whale (Balaenoptera physalus), 18 bones were identified as blue whale (B. musculus), two bones were identified as sei whale (B. borealis), one bone was identified as a southern right whale (Eubalaena australis), and one bone was identified as a southern elephant seal (Mirounga leonina). The prominence of humpback, fin, and blue whale bones in the sample collection corresponds to the catch record of the early years of whaling on the island of South Georgia (pre‐1915), prior to the depletion of these populations.     

A new species of baleen whale (Balaenoptera) from the Gulf of Mexico,with a review of its geographic distribution

Bryde's-like whales are a complex of medium-sized baleen whales that occur in tropical waters of all three major ocean basins. Currently, a single species of Bryde's whale, Balaenoptera edeni Anderson, 1879, is recognized, with two subspecies, Eden's whale, B. edeni edeni and Bryde's whale, B. edeni brydei (Olsen, 1913), although some authors have recognized these as separate species. Recently, a new, evolutionarily divergent lineage of Bryde's-like whale was identified based on genetic data and was found to be restricted primarily to the northern Gulf of Mexico (GOMx). Here, we provide the first morphological examination of a complete skull from these whales and identify diagnostic characters that distinguish it from the other medium-sized baleen whale taxa. In addition, we have increased the number of genetic samples of these Bryde's-like whales in the GOMx from 23 to 36 individuals, all of which matched the GOMx lineage. A review of Bryde's-like whale records in the Caribbean and greater Atlantic supports an isolated distribution for this unique lineage, augmenting the genetic and morphological body of evidence supporting the existence of an undescribed species of Balaenoptera from the Gulf of Mexico.     

Exposure to seismic survey alters blue whale acoustic communication

The ability to perceive biologically important sounds is critical to marine mammals, and acoustic disturbance through human-generated noise can interfere with their natural functions. Sounds from seismic surveys are intense and have peak frequency bands overlapping those used by baleen whales, but evidence of interference with baleen whale acoustic communication is sparse. Here we investigated whether blue whales (Balaenoptera musculus) changed their vocal behaviour during a seismic survey that deployed a low-medium power technology (sparker). We found that blue whales called consistently more on seismic exploration days than on non-exploration days as well as during periods within a seismic survey day when the sparker was operating. This increase was observed for the discrete, audible calls that are emitted during social encounters and feeding. This response presumably represents a compensatory behaviour to the elevated ambient noise from seismic survey operations.     

Cetacean mitochondrial DNA control region: sequences of all extant baleen whales and two sperm whale species

The sequence of the mitochondrial control region was determined in all 10 extant species commonly assigned to the suborder Mysticeti (baleen or whalebone whales) and to two odontocete (toothed whale) species (the sperm and the pygmy sperm whale). In the mysticetes, both the length and the sequence of the control region were very similar, with differences occurring primarily in the first approximately 160 bp of the 5' end of the L-strand of the region. There were marked differences between the mysticete and sperm whale sequences and also between the two sperm whales. The control region, less its variable portion, was used in a comparison including the 10 mysticete sequences plus the same region of an Antarctic minke whale specimen and the two sperm whales. The difference between the minke whales from the North Atlantic and the Antarctic was greater than that between any acknowledged species belonging to the same genus (Balaenoptera). The difference was similar to that between the families Balaenopteridae (rorquals) and Eschrichtiidae (gray whales). The findings suggest that the Antarctic minke whale should have a full species status, B. bonaerensis. Parsimony analysis separated the bowhead and the right whale (family Balaenidae) from all remaining mysticetes, including the pygmy right whale. The pygmy right whale is usually included in family Balaenidae. The analysis revealed a close relationship between the gray whale (family Eschrichtiidae) sequence and those of the rorquals (family Balaenopteridae). The gray whale was included in a clade together with the sei, Bryde's, fin, blue, and humpback whales. This clade was separated from the two minke whale types, which branched together.      

Blue and fin whale acoustic presence around Antarctica during 2003 and 2004

Seasonal and spatial variations of blue ( Balaenoptera musculus ) and fin whale ( B. physalus ) calls were analyzed from recordings collected with Acoustic Recording Packages (ARPs) deployed between January 2003 and July 2004 at four circumpolar locations: the Western Antarctic Peninsula (WAP), the Scotia Sea (SS), Eastern Antarctica (EA), and the Ross Sea (RS). Call characteristics were compared among sites using the average pressure spectrum levels from 1 month of data at each location. Presence of calls was analyzed using automatic call detection and acoustic power analysis methods. Blue whale calls were recorded year-round, with the highest detections in February–May and November. This suggests that the blue whale population may not migrate synchronously, and may indicate long duration calls are more common during migrations. Fin whale calls were detected only during February–July. Two distinct fin whale call types were recorded, suggesting a possible separation into two populations. The calls at the EA site had a secondary frequency peak in the pressure spectrum at 99 Hz and the calls at the WAP and the SS sites had a peak at 89 Hz. No fin whale calls were detected at the RS site. Acoustics are a good tool to monitor large whales in the Southern Ocean.     

Tracing the spatio-temporal dynamics of endangered fin whales (<Emphasis Type="Italic">Balaenoptera physalus</Emphasis>) within baleen whale (Mysticeti) lineages: a mitogenomic perspective

To explore the spatio-temporal dynamics of endangered fin whales (Balaenoptera physalus) within the baleen whale (Mysticeti) lineages, we analyzed 148 published mitochondrial genome sequences of baleen whales. We used a Bayesian coalescent approach as well as Bayesian inferences and maximum likelihood methods. The results showed that the fin whales had a single maternal origin, and that there is a significant correlation between geographic location and evolution of global fin whales. The most recent common female ancestor of this species lived approximately 9.88 million years ago (Mya). Here, North Pacific fin whales first appeared about 7.48 Mya, followed by a subsequent divergence in Southern Hemisphere approximately 6.63 Mya and North Atlantic about 4.42 Mya. Relatively recently, approximately 1.76 and 1.42 Mya, there were two additional occurrences of North Pacific populations; one originated from the Southern Hemisphere and the other from an uncertain location. The evolutionary rate of this species was 1.002?×?10^?3 substitutions/site/My. Our Bayesian skyline plot illustrates that the fin whale population has the rapid expansion event since ~?2.5 Mya, during the Quaternary glaciation stage. Additionally, this study indicates that the fin whale has a sister group relationship with humpback whale (Meganoptera novaeangliae) within the baleen whale lineages. Of the 16 genomic regions, NADH5 showed the most powerful signal for baleen whale phylogenetics. Interestingly, fin whales have 16 species-specific amino acid residues in eight mitochondrial genes: NADH2, COX2, COX3, ATPase6, ATPase8, NADH4, NADH5, and Cytb.     

福建平潭一头误捕灰鲸的部分形态学记录

2011 年11 月5 日,福建省平潭县白青乡青峰村渔民出海作业时发现一头误撞定置刺网死亡的灰鲸,为福建省的首次灰鲸误捕案例,也是本世纪以来我国海域的首次灰鲸误捕记录。本文描述了该灰鲸的外形特征和骨骼系统,并报道外形和骨骼的测量数据。该灰鲸系雌性,体长1 309 cm,体重约21 t,为目前本种在我国海域搁浅/ 误捕记录中的最大个体。头骨长281 cm,宽128 cm,重250 kg;脊椎式为C7 + T13 + L13 +Ca23 = 56;指式为Ⅰ1,Ⅱ3,Ⅲ7,Ⅳ5,Ⅴ3;肋骨14 对,V 形骨10 枚。脊椎式、指式、肋骨和V 形骨较之以前的报道存在差异,说明灰鲸的脊椎式、指式、肋骨和V 形骨可能存在个体差异。此外,本文还综述了西北太平洋灰鲸目前所面临的主要致危因素。     

Phenological changes in North Atlantic right whale habitat use in Massachusetts Bay

The North Atlantic right whale (Eubalaena glacialis) is one of the world's most highly endangered baleen whales, with approximately 400–450 individuals remaining. Massachusetts Bay (MB) and Cape Cod Bay (CCB) together comprise one of seven areas in the Gulf of Maine where right whales seasonally congregate. Here, we report on acoustically detected presence of right whales in MB over a nearly 6 year period, July 2007–April 2013, a time of both rapid ocean warming throughout the Gulf of Maine and apparent changes in right whale migratory dynamics. We applied an automated detection algorithm to assess hourly presence of right whale “up‐calls” in recordings from a 19‐channel acoustic array covering approximately 4,000 km² in MB. Over the survey, up‐calls were detected in 95% of 8 day periods. In each year, as expected, we observed a “peak season” of elevated up‐call detections in late winter and early spring corresponding to the season when right whales congregate to feed in CCB. However, we also saw an increase in right whale occurrence during time periods thought to be part of the “off‐season.” With the exception of 2009–2010, when acoustic presence was unusually low, the mean percent of hours in which up‐calls were detected increased every year, both during the peak season (from 38% in 2008 to 70% in 2012), and during the summer–fall season (from 2% in 2007 to 13% in 2012). Over the entire study, the peak season start date varied between 17 January and 26 February. Changes in right whale phenology in MB likely reflect broadscale changes in habitat use in other areas within the species range. This study demonstrates the value of continuous long‐term survey datasets to detect and quantify shifts in cetacean habitat use as environmental conditions change and the long‐term continued survival of right whales remains uncertain.