鳜类鱼类的线粒体DNA控制区结构及其系统发育分析

鳜类为低等鲈形目鱼类,是东亚特有类群。然而,关于其系统位置、分类以及一些物种的有效性等尚有争议。采用PCR扩增直接测序的方法,获得了鳜、大眼鳜、斑鳜、暗鳜、波纹鳜、长体鳜、中国少鳞鳜线粒体DNA控制区基因的序列。对比其他已报道鱼类控制区的结构识别序列,对鳜类鱼类控制区的结构进行了分析,识别了终止序列区、中央保守区和保守序列区,并找到了DNA复制终止相关的序列ETAS和中央保守区的保守序列CSB-F、CSB-E、CSB-D以及保守序列区的保守序列CSB1、CSB2、CSB3。几种鳜鱼间共有191个变异位点,其中,终止序列区的变异最高,占总变异的61．3％,中央保守区和保守序列区占总变异的38．7％。这一结果可为全面了解鱼类线粒体DNA控制区的结构特征提供资料。同时,利用高度变异的控制区序列,以鲈科和错科作为外群,使用邻接法和最大简约法构建了这几种鳜鱼的系统发育树。结果表明：鳜类为一单系类群,鳜、大眼鳜、斑鳜、暗鳜、波纹鳜、长体鳜构成一支鳜鱼群,其中,鳜与大眼鳜为姐妹种;中国少鳞鳜为另一支少鳞鳜群;长体鳜未单独成一支,而是聚入鳜鱼群内,应更名为Siniperca roulei。研究结果支持将现生鳜类分为两个类群的观点。     

鲹科鱼类线粒体DNA控制区结构及系统发育关系

采用PCR技术获得了9种鲹科鱼类的线粒体DNA控制区全序列,并结合从GenBank中下载的3种鲹科鱼类的相应序列采用ClustalW排序后,对控制区结构进行分析,识别了其终止序列区、中央保守区和保守序列区3个区域,指出了终止相关序列的主体是TACAT与其反向互补序列ATGTA以及一系列保守序列(CSB-F、CSB-E、CSB-D和CSB-1、CSB-2、CSB-3),并给出了它们的一般形式,同时在康氏似鲹控制区的5′和3′两端发现重复序列。以尖吻鲈作为外类群,应用邻接法构建的分子系统树表明:鲹科鱼类分为鲹亚科、鰤亚科、鲳鲹亚科和鰆鲹亚科4个亚科,各自形成单系群;鲹亚科与鰤亚科形成姐妹群,鲳鲹亚科再与他们聚在一起,鰆鲹亚科处于鲹科鱼类的基部,与前面3个亚科聚在一起。     

鲹科鱼类线粒体DNA控制区结构及系统发育关系

采用PCR技术获得了9种鲹科鱼类的线粒体DNA控制区全序列,并结合从GenBank中下载的3种鲹科鱼类的相应序列采用Clustal W排序后,对控制区结构进行分析,识别了其终止序列区、中央保守区和保守序列区3个区域,指出了终止相关序列的主体是TACAT与其反向互补序列ATGTA以及一系列保守序列（CSB-F、CSB-E、CSB-D和CSB-1、CSB-2、CSB-3）,并给出了它们的一般形式,同时在康氏似鲹控制区的5′和3′两端发现重复序列。以尖吻鲈作为外类群,应用邻接法构建的分子系统树表明：鲹科鱼类分为鲹亚科、鰤亚科、鲳鲹亚科和鰆鲹亚科4个亚科,各自形成单系群;鲹亚科与鰤亚科形成姐妹群,鲳鲹亚科再与他们聚在一起,鰆鲹亚科处于鲹科鱼类的基部,与前面3个亚科聚在一起。     

沙鳅亚科鱼类线粒体DNA控制区结构分析及系统发育关系的研究

对沙鳅亚科鱼类3属14个代表种的线粒体DNA控制区序列的结构进行了分析。通过与鲤形目鱼类的控制区序列进行比较,将沙鳅亚科鱼类的控制区分为终止序列区、中央保守区和保守序列区三个区域。同时识别了沙鳅亚科中一系列保守序列,并给出了它们的一般形式。以胭脂鱼为外类群,对比条鳅亚科、花鳅亚科、以及平鳍鳅科的代表性种类,采用NJ、MP和ML法构建沙鳅亚科的分子系统树。分子系统发育分析表明,沙鳅亚科为一单系,包括3个属:沙鳅属、副沙鳅属和薄鳅属,各属均构成单系。根据分子系统学、形态学的结果及地理分布推断,沙鳅亚科中沙鳅属可能为最为原始的属,副沙鳅属其次,而薄鳅属最特化。       

虾虎鱼类线粒体全基因组序列结构特征分析及系统发育关系探讨

虾虎鱼类体态变异大、体型小、种类多, 形态鉴定及谱系分类较为困难。为深入开展虾虎鱼类的鉴定、分类及遗传进化等研究, 文章对已获得的26种虾虎鱼线粒体全基因组进行分析。结果发现, 虾虎鱼类线粒体基因组的基因组成及排列模式与大多数脊椎动物线粒体基因组特征基本一致; 由于不同物种的控制区存在不同数量的重复序列而导致基因组序列长度存在明显的差异; 26种虾虎鱼线粒体全基因组序列及不同基因中A+T的含量均超过50%, 并存在碱基G偏倚现象。基于37个编码基因序列, 利用Kimura双参数法计算遗传距离, 发现矛尾刺虾虎鱼与斑尾刺虾虎鱼、斑纹舌虾虎鱼与钝吻舌虾虎鱼分别为同种异名。通过对26种虾虎鱼线粒体基因组控制区序列的比较, 识别了终止结合序列区、中央保守区及保守序列区。利用26种虾虎鱼线粒体基因组的36个编码基因序列构建系统发育树, 发现部分聚类结果不同于传统的形态学分类方式, 虾虎鱼科中的5个亚科出现了明显的分化, 近盲虾虎鱼亚科、背眼虾虎鱼亚科、瓢虾虎鱼亚科亲缘关系较近而聚成一大支, 然后与拟虾虎鱼亚科种类形成姐妹类群, 虾虎鱼亚科与其它的4个亚科亲缘关系较远, 单独成为一个类群。根据分子钟估算结果推测虾虎鱼科物种可能起源于始新世晚期至渐新世时段, 在中新世进一步分化为具有现代表征的虾虎鱼种类。     

圆斑星鲽及相关种类线粒体DNA控制区结构分析

采用PCR产物直接测序法测定了圆斑星鲽(Verasper variegatus)的24个个体的线粒体控制区(Control region)核苷酸全序列, 并进行了结构分析。结果表明, 圆斑星鲽线粒体控制区核苷酸全序列具有长度多态性, 得到4种长度单元型, 主要表现为控制区中的串联重复序列的长度不同。对鲽形目鱼类如鲽科的条斑星鲽(Verasper moseri)、黄盖鲽(Limanda feruginea)、马舌鲽 (Reinhardtius hippoglossoides), 美洲拟庸鲽(Heppoglossoides platessoides )和鲆科的牙鲆(Paralichthys olivaceus)以及鳎科的欧洲鳎(Solea solea)、塞内加尔鳎(S. senegalensis)和沙鳎(S. lascari)的控制区的比较研究发现, 鲽形目鱼类的线粒体控制区均存在相似的结构, 即线粒体控制区可分为终止相关序列区(ETAS)、中央保守区(包括CSB-A、CSB-B、CSB-C、CSB-D、CSB-E、CSB-F)以及保守序列区(CSB1、CSB2、CSB3)和重复序列区(Repeat region)4个区域。通过与脊椎动物各个纲线粒体控制区序列的比较分析, 发现只有鲽形目(包括鲆、鲽类和鳎类)鱼类和两栖纲的无尾类在CSB-3之后存在相似的串联重复序列。     

中国菌蚊科属的系统发育关系分析(双翅目:眼菌蚊总科)

采用Hennig 86程序,以柄菌蚊科和喙菌蚊科代表种为外群,选取48个特征,使用mhen-nig^*和bb^*指令在586微机上运算,首次对菌蚊科中5亚科28属的28种进行支序分析,探讨各分类单元系统发育关系。结果表明：菌蚊亚科与滑菌蚊亚科的亲缘关系较近,二者互为姐妹群,粘菌蚊亚科属于原始类群;菌蚊亚科为5个亚科中的进化类群;邻菌蚊亚科可能为并系群;真菌蚊亚科是介于邻菌蚊亚科与菌蚊亚科之间的类群。     

基于12S rRNA和16S rRNA基因序列探讨中国蚤蝇科部分属间的系统发育关系

基于线粒体12S rRNA和16S rRNA基因序列联合分析,采用最大简约法、最大似然法和贝叶斯法分别构建了中国蚤蝇科14属的系统发育树.结果表明:联合分析序列总长度为819 bp,其中可变位点277个,简约信息位点200个;A+T平均含量为77.7%,具A、T偏倚性.系统发育分析显:中国蚤蝇科为单系发生,分为蚤蝇亚科和裂蚤蝇亚科两个单系群.蚤蝇亚科内脉蚤蝇属、锥蚤蝇属和刺蚤蝇属亲缘关系较近,栅蚤蝇属与栓蚤蝇属亲缘关系较近;裂蚤蝇亚科中虼蚤蝇属与裂蚤蝇属互为姐妹群,寡蚤蝇属与伐蚤蝇属互为姐妹群.     

印度谷螟线粒体基因组测定及螟蛾总科系统发育分析

线粒体基因组广泛应用于昆虫系统进化、种群遗传学及谱系地理学等众多研究领域.印度谷螟(Plodia interpunctella)属鳞翅目螟蛾科,是世界性重要仓储害虫.为了深入了解螟蛾总科昆虫的系统进化关系,本研究测定并分析了印度谷螟的全线粒体基因组.印度谷螟线粒体基因组全长15264 bp,包括13个蛋白编码基因、22个转运RNA基因、2个核糖体RNA基因和一个大的非编码区(线粒体控制区).比较线粒体基因组学分析表明,线粒体基因组的基因排序、碱基组成、密码子使用、控制区的序列结构以及转运RNA基因的二级结构,在已测的25种螟蛾总科昆虫中高度保守.线粒体蛋白编码基因的进化模式分析表明,13个基因的进化速率在螟蛾科和草螟科间存在明显差异.基于13个蛋白编码基因对螟蛾总科的9个亚科的系统发育分析显示,贝叶斯树和最大似然树均高度支持螟蛾科和草螟科为2个单系群.在螟蛾科中,斑螟亚科与丛螟亚科具有更近的亲缘关系.草螟科包括2个分支,一支由野螟亚科和斑野螟亚科构成,另一支包括草螟亚科、水螟亚科、苔螟亚科和禾螟亚科等4个亚科.     

Phylogenetic relationships and divergence times among mustelids (Mammalia: Carnivora) based on nucleotide sequences of the nuclear interphotoreceptor retinoid binding protein and mitochondrial cytochrome b genes

Phylogenetic relationships among 20 species-group taxa of Mustelidae, representing Mustelinae (Mustela, Martes, Gulo), Lutrinae (Enhydra), and Melinae (Meles), were examined using nucleotide sequences of the nuclear interphotoreceptor retinoid binding protein (IRBP) and mitochondrial cytochrome b genes. Neighbor-joining and maximum-parsimony phylogenetic analyses on these genes separately and combined were conducted. While IRBP performed better than cytochrome b in recovering more-inclusive clades, cytochrome b demonstrated more resolving power in recovering less-inclusive clades. Strong support was found for a close affinity of Enhydra with Mustela to the exclusion of Martes and Gulo (causing Mustelinae to be paraphyletic); the most-basal position of Mustela vison within Mustela, followed by Mustela erminea; an association of Mustela lutreola, Mustela itatsi, Mustela sibirica, and the subgenus Putorius (including Mustela putorius and Mustela eversmanii), to the exclusion of Mustela nivalis and Mustela altaica; and a basal position of Mustela itatsi to a clade containing Mustela sibirica and Putorius. Whereas cytochrome b strongly supported Mustela lutreola as the sister species to Putorius, IRBP strongly supported its basal placement to the Mustela itatsi-Mustela sibirica-Putorius clade. The low level of sequence divergence in cytochrome b between Mustela lutreola and Putorius is therefore a result of interspecific mitochondrial introgression between these taxa, rather than a recent origin of Mustela lutreola in a close relationship to Putorius. Time estimates inferred from IRBP and cytochrome b for mustelid divergence events are mostly in agreement with the fossil record.     

A comparative analysis of the complete mitochondrial genome of the Eurasian otter <Emphasis Type="Italic">Lutra lutra</Emphasis> (Carnivora; Mustelidae)

Otter populations are declining throughout the world and most otter species are considered endangered. Molecular methods are suitable tools for population genetic research on endangered species. In the present study, we analyzed the complete mitochondrial genome (mitogenome) sequence of the Eurasian otter Lutra lutra. The mitochondrial DNA sequence of the Eurasian otter is 16,505 bp in length and consists of 13 protein-coding genes, 22 tRNAs, 2 rRNAs, and a control region (CR). The CR sequence of otters from Europe and Asia showed nearly identical numbers and nucleotide sequences of minisatellites. Phylogenetic analysis of Mustelidae mitogenomes, including individual genes, revealed that Lutrinae and Mustelinae form a clade, and that L. lutra and Enhydra lutris are sister taxa within the Lutrinae. Phylogenetic analyses revealed that of the 13 mitochondrial protein-coding genes, ND5 is the most reliable marker for analysis of phylogenetic relationships within the Mustelidae.     

New insights into the evolution of intronic sequences of the beta-fibrinogen gene and their application in reconstructing mustelid phylogeny

Mustelidae is the largest and most diverse family in the order Carnivora. The phylogenetic relationships among the subfamilies have especially long been a focus of study. Herein we are among the first to employ two new introns (4 and 7) of the nuclear beta-fibrinogen gene to clarify these enigmatic problems. In addition, two previously available nuclear (IRBP exon 1 and TTR intron 1) and one mt (ND2) data sets were also combined and analyzed simultaneously with the newly obtained sequence data in this study. Detailed characterizations of the two intronic regions not only reveal the remarkable occurrences of short interspersed element (SINE) insertion events, providing a new example supporting the attractive hypothesis that attrition of an earlier retroposition may offer a proper environment for successive retropositions by forming a "dimer-like" structure, but also demonstrate their utility in the resolution of mustelid phylogeny. All of our analyses confirm the assemblage of Mustelinae, Lutrinae, and Melinae with confidence; moreover, two clades within Mustelinae were clearly recognized, i.e., genera Mustela and Martes. Notably, genus Martes of Mustelinae was found to branch off first, followed by Melinae and then a clade containing Lutrinae and genus Mustela of Mustelinae, indicating paraphyly of Mustelinae. In addition, Mephitinae diverges before the other mustelids and the monophyletic Procyonidae in all cases, supporting its elevation to a separate family. Additional independent genetic markers are still in need to resolve the trichotomy among Mephitinae and the other two carnivoran clades, Ailuridae and Procyonidae/non-mephitine Mustelidae.     

Molecular phylogeny of arctoids (Mammalia: Carnivora) with emphasis on phylogenetic and taxonomic positions of the ferret-badgers and skunks

Phylogenetic relationships among the ferret-badger Melogale moschata, the skunk Mephitis mephitis, and 21 other arctoid carnivorans, representing Mustelidae (Mustelinae: Mustela, Martes, Gulo; Lutrinae: Enhydra; Melinae: Meles), Procyonidae (Procyon), and Ursidae (Ursus, Melursus), were evaluated through maximum-parsimony phylogenetic analysis of concatenated partial nucleotide sequences of the nuclear recombination-activating gene 1 (RAG1) and gene encoding interphotoreceptor retinoid-binding protein (IRBP). The analysis strongly supports Melogale as more closely related to a musteline-lutrine clade (containing Mustela and Enhydra) than to Meles or another musteline clade containing Martes and Gulo (causing Melinae and Mustelinae, as traditionally circumscribed, to be nonmonophyletic). This, together with known morphological and karyological evidence for nonmeline affinities of Melogale, justify the exclusion of the ferret-badgers from the monophyletic Melinae. Therefore, we recommend that Melogale be classified in a distinct mustelid subfamily, the monotypic Helictidinae. Our analysis also strongly supports an outgroup position of the skunks to a clade containing Procyonidae and the nonmephitine Mustelidae (causing Mustelidae, as traditionally circumscribed, to be paraphyletic). This position of the skunks agrees with results of most previous genetic studies. However, it is contradicted by known morphological evidence from both living and fossil taxa, as well as genetic evidence from protein electrophoresis. These consistently support the traditional placement of the skunks within the monophyletic Mustelidae (recently in a close relationship to Lutrinae). Therefore, we consider the recent elevation of the skunks to the level of family as premature, and recommend that this clade be left at the subfamily level (Mephitinae) within the family Mustelidae, pending further evidence.     

南蝠线粒体DNA控制区结构及贵州7个种群遗传多样性的分析研究

采用PCR技术获得了贵州7个南蝠(Ia io)自然种群42个个体的线粒体DNA控制区全序列,长度为1256～1340 bp.对控制区结构进行分析,识别了其延伸的终止结合序列区(包括ETAS1和ETAS2元件)、中央保守区(包括F、E、D、C、B元件)和保守序列区(包括CSB1、CSB2和CSB3元件);同时,在延伸的终止结合序列区还发现了若干能形成发夹结构的主体序列TACAT—ATGTA.在7个自然种群42个个体中共定义了16个单倍型.遗传多样性分析表明:贵州南蝠种群具有较高的单倍型多样性(h=0.945)和中等的核苷酸多样性(π=0.012).基因流、AMOVA和系统进化树分析表明贵州这7个南蝠自然群体间没有发生遗传分化.     

DNA sequence variation in the mitochondrial control region of subterranean mole rats,Spalax ehrenbergi superspecies,in Israel

The complete mitochondrial control region was sequenced for 60 individuals representing different populations for each of the four species of the subterranean mole rat Spalax ehrenbergi superspecies in Israel: Spalax galili (2n = 52), S. golani (2n = 54), S. carmeli (2n = 58), and S. judaei (2n = 60). The control region of all species and populations is very similar both in length (979 to 983 bp) and in base composition. As in agreement with previous surveys on mitochondrial control regions on mammals, the mole rat control region can be divided into a central domain and two flanking domains, ETAS (extended termination associated sequences) and CSB (conserved sequence blocks). Along with the common conserved blocks found in these domains (ETAS1, ETAS2, CSB1, CSB2, and CSB3), we have also detected in all individuals an ETAS1-like and a CSB1-like element, both in the ETAS domain. The most conserved region was the central domain, followed by the CSB and ETAS domains, showing important differences in the four species analyzed. Phylogenetic analysis supported the existence of two clades. One clade contained individuals belonging to Spalax galili (2n = 52) and S. golani (2n = 54), separated in two different branches depending on the species. The other clade contained individuals belonging to S. carmeli (2n = 58) and S. judaei (2n = 60) mixed together, suggesting a more recent event of speciation. Within species we have observed a southward trend of increasing variability. These results have been explained as a consequence of the adaptation of the species to ecological factors such as aridity and temperature stresses.     

Structure,DNA sequence variation and phylogenetic implications of the mitochondrial control region in horseshoe bats

There have been few studies of the structural and evolutionary characteristics of the mitochondrial control region (CR) in rhinolophids, yet this could have important consequences for the interpretation of phylogenetic relationships within this group. Here we sequenced and analyzed the CR of 37 individuals from 12 Rhinolophus species, including 2 species from GenBank. The length of the CR ranged from 1335 to 1514 bp, and the base composition was very similar among species. The CR of horseshoe bats, like that of other mammals, could be subdivided into a central conserved domain (CD) and two flanking variable domains, extended termination associated sequences (ETAS), and conserved sequence blocks (CSB). Besides the common conserved blocks (ETAS1, ETAS2, F-B boxes, CSB1, CSB2 and CSB3) found in 3 domains, an ETAS2-like and a CSB1-like element were also detected in the ETAS and CSB domains, respectively, in all individuals. Notwithstanding a short tandem repeat (11 or 13 bp) between CSB1 and CSB2 in all specimens, the base composition, copy number and arrays are all variable. A long tandem repeat (79 bp) was only identified in the ETAS domain in one individual of R. pusillus. Phylogenetic reconstructions based on the CR sequences indicated that the molecular phylogenetic relationships among some Rhinolophus species were inconsistent with the results of phenetic analyses, but similar to phylogenetic constructions using cytochrome b. An unidentified species R. sp and 3 species from the philippinensis-group that were clearly morphologically different comprised a monophyletic group, which could have resulted from morphological independent evolution.     

Evolutionary trends of the mitochondrial lineage differentiation in species of genera Martes and Mustela

We compared partial sequences (402 bp) of the mitochondrial cytochrome b gene in 68 individuals of martens (Martes), weasels (Mustela) and their relatives from the Northern Hemisphere to identify the modes of geographic differentiation in each species. We then compared complete sequences (1140 bp) of the gene in 17 species of the family Mustelidae to know the spatial and temporal modes of speciation, constructing linearized trees with transversional substitutions for deeper lineage divergences and with transversions and transitions for younger lineages. Our data suggested that these lineages of Martes and Mustela differentiated in a stepwise fashion with five radiation stages from the generic divergences (stage I) to the intraspecific divergences (stage V), during the last 10 or 20 million years as the fossil evidence suggests. In the lineage of Martes, the first offshoots are of Martes flavigula, M. pennanti, and Gulo gulo (stage II), the second is M. foina (stage III), and the third are M. americana, M. martes, M. melampus, and M. zibellina (stage IV). The divergence of the lineages of Mustela is likely to have taken place concurrently with the radiations of the Martes. These divergence processes are attributable in part to the geographic allocation along the two continents, North America and Eurasia, as well as among peripheral insular domains, such as Taiwan and the Japanese Islands. In addition, the Eurasian continent itself was shown to have been involved in the species diversification in the martens and weasels.     

DNA sequence variation in the mitochondrial control region of red-backed voles (Clethrionomys)

The complete mitochondrial DNA (mtDNA) control region was sequenced for 71 individuals from five species of the rodent genus Clethrionomys both to understand patterns of variation and to explore the existence of previously described domains and other elements. Among species, the control region ranged from 942 to 971 bp in length. Our data were compatible with the proposal of three domains (extended terminal associated sequences [ETAS], central, conserved sequence blocks [CSB]) within the control region. The most conserved region in the control region was the central domain (12% of nucleotide positions variable), whereas in the ETAS and CSB domains, 22% and 40% of nucleotide positions were variable, respectively. Tandem repeats were encountered only in the ETAS domain of Clethrionomys rufocanus. This tandem repeat found in C. rufocanus was 24 bp in length and was located at the 5' end of the control region. Only two of the proposed CSB and ETAS elements appeared to be supported by our data; however, a "CSB1-like" element was also documented in the ETAS domain.     

Fruit consumption by carnivores in Mediterranean Europe

• 1 We reviewed patterns of fruit consumption amongst 10 species of mesocarnivores: red fox Vulpes vulpes, weasel Mustela nivalis, stoat Mustela erminea, polecat Mustela putorius, stone marten Martes foina, pine marten Martes martes, Eurasian badger Meles meles, common genet Genetta genetta, Egyptian mongoose Herpestes ichneumon and wildcat Felis silvestris in Mediterranean Europe.
• 2 The 65 reviewed studies recorded 79 different fruits eaten by carnivores, 58 of which were identified to species. Most records (63%) were of fleshy fruits with high pulp content. The frequency of occurrence of fruit items varied widely amongst species and regions. Four of the carnivore species (red fox, stone marten, badger and common genet) included more than 30 fruit species in their diet.
• 3 A longitudinal pattern was detected in the consumption of fruit in the Mediterranean region, with the frequency of occurrence of fruit consumption increasing towards the east.