Direito Desportivo Internacional: a influência das normas e das decisões jusdesportivas internacionais no ordenamento jurídico brasileiro

Este trabalho tem como escopo analisar as influências das normas e das decisões jusdesportivas internacionais no ordenamento jurídico brasileiro. Para isso, alinharam-se as predileções do autor por Esporte, Direito Internacional e Relações Internacionais, assim como por outras ciências, como Geografia. Indubitável a importância do tema para a conjuntura atual, sendo o Brasil sede dos dois principais megaeventos esportivos em 2014 e 2016, a Copa do Mundo Fifa e os Jogos da XXXI Olimpíadas, os Jogos Olímpicos. Essa dissertação foi dividida em quatro capítulos. O primeiro trata da relação entre Esporte, Globalização e Geopolítica, e o segundo capítulo versa sobre o Esporte sob a perspectiva dos Direitos Humanos. Após estudar o Esporte sob a perspectiva do Estado e do Homem, analisam-se as fontes do Direito Desportivo Internacional e o ordenamento jusdesportivo. Os Estados não são os únicos produtores de normas jurídicas. Entidades privadas são capazes de produzir normas, em cooperação ou em conflito com o Estado. Admite-se, portanto, pluralidade de produção normativa. No caso do Direito Desportivo Internacional, optou-se por abordar quatro organizações esportivas internacionais: COI, Wada, Fifa e TAS/CAS, tema do derradeiro capítulo. A influência de seus regulamentos e de seus estatutos se mostrará não apenas nos estatutos e nos regulamentos de suas afiliadas, mas também na legislação nacional. Essas organizações não governamentais de alcance transnacional/internacional têm sua natureza jurídica de acordo com seus estatutos constitutivos. A tentativa de unificação de sistemas jurídicos deve ser abandonada, dando lugar à concepção de harmonização. A dificuldade de harmonização é causada pela pluralidade de Estados e de atores privados que tentam normatizar as relações jurídicas que envolvem o esporte. Desse modo, as influências dessas normas internacionais de caráter privado são marcantes no ordenamento jurídico pátrio. Analisa-se como ocorrem essas influências e de que forma cada produtor de norma jurídica poderia atuar para que prevaleça a cooperação entre eles no âmbito jusdesportivo.

Estrutura populacional e filogeografia de Panulirus argus (Latreille, 1804)

Panulirus argus (Latreille, 1804) é uma das principais espécies de lagosta no Atlântico, sendo um dos maiores recursos pesqueiros do Atlântico Ocidental, onde apresenta um alto valor comercial. A forte explotação da espécie resulta em uma grande pressão sobre suas populações. Recentemente, foi descoberto que sob o binômio P. argus estão contidas duas espécies crípticas que ocorrem em alopatria, uma na região do Caribe e outra na costa brasileira. Esta tese tem como objetivo estudar como se estruturam geneticamente as populações dessas duas espécies, com o propósito de fornecer mais informações para a determinação de estoques e um correto manejo das espécies, e analisar os processos históricos evolutivos que moldaram suas histórias demográficas. Para tal, foram estudados dois marcadores mitocondriais (região controle e o gene da Citocromo Oxidase I) e loci de microssatélites de indivíduos de 7 regiões do Caribe (Florida, Bahamas, Turks e Caicos, Porto Rico, Cuba, Colômbia e Venezuela) e 11 estados do Brasil (Pará, Maranhão, Piauí, Ceará, Rio Grande do Norte, Pernambuco, Alagoas, Bahia, Espírito Santo, Rio de Janeiro e São Paulo). Dentro de cada espécie foram observadas duas linhagens mitocondriais diferentes, que co-ocorriam, de maneira homogênea, ao longo de suas distribuições. Hipotetiso que essas linhagens foram formadas a partir de um evento de vicariância com contato secundário ou como consequência de um efeito gargalo seguido de expansão. As duas linhagens são evidentes nas sequências da região controle mitocondrial, mas no gene da COI foram evidentes apenas em P. cf. argus do Caribe. As linhagens do Brasil se separaram há aproximadamente 233 - 288 mil anos e cada uma sofreu expansão em tempos diferentes, a primeira se expandiu há 100 mil anos e a segunda linhagem há 50 mil anos. As linhagens do Caribe se separaram cerca de 1 milhão de anos atrás e possuem o mesmo tempo de expansão, 50 mil anos. Os microssatélites não revelaram subdivisão populacional para nenhuma das duas espécies, porém os marcadores, juntos, sugeriram um fluxo gênico diferenciado entre localidades expostas a diferentes correntes marítimas. Considerando que essas lagostas são intensamente explotadas, é importante ser cuidadoso no momento de definir estoques pesqueiros. Para a espécie do Brasil, dois estoques pesqueiros foram sugeridos, o primeiro do Pará à Bahia e o segundo do sul da Bahia a São Paulo. Para a espécie do Caribe, foi mantida e reforçada a hipótese de quatro estoques sugerida pela FAO (Norte, Sul, Centro-Norte e Centro-Sul).

A molecular genetic investigation of the population structure of Atlantic menhaden (Brevoortia tyrannus)

Atlantic menhaden (Brevoortia tyrannus), through landings, support one of the largest commercial fisheries in the United States. Recent consolidation of the once coast-wide reduction fishery to waters within and around Chesapeake Bay has raised concerns over the possibility of the loss of unique genetic variation resulting from concentrated fishing pressure. To address this question, we surveyed variation at the mitochondrial cytochrome c oxidase subunit I (COI) gene region and seven nuclear microsatellite loci to evaluate stock structure of Atlantic menhaden. Samples were collected from up to three cohorts of Atlantic menhaden at four geographic locations along the U.S. Atlantic coast in 2006 and 2007, and from the closely related Gulf menhaden (B. patronus) in the Gulf of Mexico. Genetic divergence between Atlantic menhaden and Gulf menhaden, based on the COI gene region sequences and microsatellite loci, was more characteristic of conspecific populations than separate species. Hierarchical analyses of molecular variance indicated a homogeneous distribution of genetic variation within Atlantic menhaden. No significant variation was found between young-of-the-year menhaden (YOY) collected early and late in the season within Chesapeake Bay, between young-of-the-year and yearling menhaden collected in the Chesapeake Bay during the same year, between YOY and yearling menhaden taken in Chesapeake Bay in successive years, or among combined YOY and yearling Atlantic menhaden collected in both years from the four geographic locations. The genetic connectivity between the regional collections indicates that the concentration of fishing pressure in and around Chesapeake Bay will not result in a significant loss of unique genetic variation.

Filogeografia e evolução molecular de lagostas sapateiras (Scyllarides spp.) da América do Sul

Nos últimos anos, duas espécies de lagostas sapateiras, Scyllarides brasiliensis e S. deceptor, vêm se destacando nos desembarques pesqueiros de lagostas do Atlântico Sul Ocidental. Para espécies comercialmente importantes, o desenvolvimento de estudos que permitam conhecer a variabilidade e entender a dinâmica populacional é fundamental. Assim, o objetivo do primeiro capítulo desta tese foi avaliar a diversidade genética e a estrutura populacional dessas duas lagostas ao longo de aprox. 2.800 km da costa da América do Sul. Para as análises, foram empregados marcadores mitocondriais (citocromo oxidase I: COI; e a região controle: RC) e marcadores nucleares (13 loci de microssatélites desenvolvidos nesta tese). As duas espécies apresentaram altos níveis de variabilidade (S. deceptor: N = 200, mtDNA: h > 0,841, π > 0,005; microssatélites: He = 0,685; S. brasiliensis: N = 211, He = 0,554), distribuídos homogeneamente entre as localidades (S. deceptor: ΦST < -0,004, ΦCT < 0,016, FST global = 0,001, Dest global = 0,003, FCT < 0,002, P > 0,05, K = 1; S. brasiliensis: FST global = 0,004, Dest global = 0,001, FCT < 0,004, P > 0,05, K = 1). A ausência de estruturação nas duas espécies pode estar relacionada a características biológicas que promovem a conectividade entre localidades geograficamente distantes, como alta fecundidade e alto potencial de dispersão das larvas planctônicas. Além disso, os dados mitocondriais sugerem que a história demográfica de S. deceptor foi marcada por eventos de expansão populacionais e geográficos possivelmente relacionados às condições ambientais favoráveis dos episódios interglaciais do Pleistoceno Médio-Tardio. Diversos estudos têm mostrado que os fenômenos de inserção de regiões mitocondriais no DNA nuclear (NuMts) e heteroplasmia limitam a correta amplificação e identificação dos marcadores mitocondriais. Em estudos filogenéticos e de genética de populações, a presença inadvertida de sequências de diversas origens viola o principio de ortologia, o que pode resultar em inferências evolutivas erradas. Assim, o objetivo do segundo capítulo desta tese foi identificar e caracterizar os possíveis NuMts e sequências heteroplásmicas de três regiões mitocondriais (COI, RC e o gene da subunidade maior do RNA ribossomal: 16S) em quatro espécies do gênero Scyllarides (S. aequinoctialis, S. brasiliensis, S. deceptor e S. delfosi). A clonagem e sequenciamento de extratos de DNA genômico e DNA enriquecido com mtDNA revelaram que os genomas destas espécies podem exibir NuMts (que divergem entre 0,6 e 17,6% do mtDNA) e heteroplasmia (que divergem < 0,2% do mtDNA prevalente). Os NuMts surgiram possivelmente de vários eventos independentes de integração ao núcleo ao longo da história evolutiva do gênero Scyllarides. Dependendo do seu grau de similaridade com o mtDNA, a presença de NuMts nas análises filogenéticas no nível de gênero pode causar superestimativa do número de espécies e alterações nos comprimentos dos ramos e nas relações filogenéticas entre espécies.

A new species of western Atlantic lizardfish (Teleostei: Synodontidae: Synodus) and resurrection of Synodus bondi Fowler, 1939, as a valid species from the Caribbean with redescriptions of S. bondi, S. foetens (Linnaeus, 1766), and S. intermedius (Agassiz, 1829)

Western Atlantic synodontid species were studied as part of an ongoing effort to reanalyze Caribbean shorefish diversity. A neighbor-joining tree constructed from cytochrome c oxidase I (COI) data revealed 2 highly divergent genetic lineages within both Synodus intermedius (Agassiz, 1829) (Sand Diver) and S. foetens (Linnaeus, 1766) (Inshore Lizardfish). A new species, Synodus macrostigmus, is described for one of the S. intermedius lineages. Synodus macrostigmus and S. intermedius differ in number of lateral-line scales, caudal pigmentation, size of the scapular blotch, and shape of the anterior-nostril flap. Synodus macrostigmus and S. intermedius have overlapping geographic and depth distributions, but S. macrostigmus generally inhabits deeper water (>28 m) than does S. intermedius and is known only from coastal waters of the southeastern United States and the Gulf of Mexico, in contrast to those areas and the Caribbean for S. intermedius. Synodus bondi Fowler, 1939, is resurrected from the synonymy of S. foetens for one of the S. foetens genetic lineages. The 2 species differ in length and shape of the snout, number of anal-fin rays, and shape of the anterior-nostril flap. Synodus bondi and S. foetens co-occur in the central Caribbean, but S. bondi otherwise has a more southerly distribution than does S. foetens. Redescriptions are provided for S. intermedius, S. foetens, and S. bondi. Neotypes are designated for S. intermedius and S. foetens. A revised key to Synodus species in the western Atlantic is presented.

云南闭壳龟(Cuora yunnanensis)的分子鉴定及进化地位研究

云南闭壳龟(Cuora yunnanensis,Boulenger,1906)曾被认为已经灭绝,在保护生物学上受到广泛的关注.本文测定了3只活的云南闭壳龟线粒体COI和ND4的部分序列及His,Ser,Leu tRNA序列片段,共 1725 个碱基序列.结合闭壳龟属其他物种序列,包括之前测定的一只云南闭壳龟标本(MNHN 1907.10)的DNA序列,进行了分子系统学分析.与100年前的标本比较,无论是形态还是分子系统分析结果都显示,这种新发现的活龟确实是云南闭壳龟.同时,我们的结果确证了标本序列的可信性,揭示云南闭壳龟不是近期杂交形成的,而是代表了进化上独立的遗传谱系,且种内仍存在一定的遗传多样性.

Phylogenetic relationships of 12 Penaeoidea shrimp species deduced from mitochondrial DNA sequences

DNA sequences of an 847 bp fragment of mitochondrial cytochrome oxidase subunit I (COI) gene and a 514 bp fragment of 16s rRNA gene were determined to examine the phylogenetic relationships of 12 Penaeoidea shrimp species (Penaeus chinensis, Penaeus japon

Study morphologic, phylogenic relationships DNA barcoding of of identified decapod group (Crustacean) of Persian Gulf and Oman Sea.

Phylogenetic relationships among all described species (total of 12 taxa) of the decapoda, were examined with nucleotide sequence data from portions of mitochondrial gene and cytochrome oxidase subunit I (COI). The previous works on phylogeny proved that the mitochondrial COI gene in crustacean is a good discriminative marker at both inter- and intra-specific levels. We provide COI barcode sequences of commertial decapoda of Oman Sea, Persian Gulf, Iran. Industrial activities, ecologic considerations, and goals of the decapoda Barcode of Life campaign make it crucial that species of the south costal be identified. The reconstruction of evolut phylogeny of these species are crucial for revealing stock identity that can be used for the management of fisheries industries in Iran. Mitochondrial DNA sequences were used to reconstruct the phylogeny of the Penaeus species of marine shrimp. For this purpose, DNA was extracted using phenol- chloroform well as CTAB method. The evolutionary relationships among 12 species of the decapoda were examined using 610 bp of mitochondrial (mt) DNA from the cytochrome oxidase subunit I gene. Finally the cladograms were compared and the resulted phylogenetic trees confirmed that the Iran's species origin is Indo-west pacific species. Iran's species, which were not grouped with the other decapoda taxa seem to always form a sister clade with Indo-west pacific species with strong bootstrap support 100%. The result completely agrees with the previously defined species using morphological characters.However, we still lack any comprehensive and clear understanding of phylogenetic relationships in this group.

A survey on relation of morphological, molecular and phylogenetic structure of zoanthids of the islands located in the Hormoz Strait (Hormoz, Qeshm, Larak, Hengam)

The order Zoantharia (Zoanthids) is one of the most neglected orders of cnidarians in the Persian Gulf. The present study aims to investigate the biodiversity of this order with morphological and molecular examination in the Persian Gulf. For this purpose, 123 colonies of zoanthids with variety of shape and colors have been collected from intertidal and shallow water zone of four islands, i. e. Hengam, Qeshm, Larak and Hormoz. After sampling, morphological characteristics of each specimen were recorded based on in situ photographs. Then DNA was extracted using the cetyl trimethyl ammonium bromide (CTAB) method. Both mitochondrial 16S ribosomal DNA (mt 16S rDNA) and cytochrome oxidase subunit I (COI) gene fragments were amplified and sequenced. The results of preliminary morphological identification integrated with two mitochondrial markers sequencing demonstrated the presence of five different species in this region; Zoanthus sansibaricus, Palythoa mutuki, Palythoa cf. mutuki, Palythoa tuberculosa and Neozoanthus persicus?. Although at first sight, morphological properties were not successful to delineate zoanthid species, they become reliable criteria to identify and delineate species in field studies after molecular identification.

Investigation of polymorphism by PCR-RFLP method in Cobia fish Rachycentron canadum in the Persian Gulf and Oman Sea

Cobia is a native fish species in Iranian waters in the Persian Gulf and Sea of Oman and has a good internal and foreign market. This fish is a fast growing species and for this reason Iranian Fisheries is considering to go for it culture practices. To go for any utilization such as fishing from wild stocks or culture activities, needs a better understanding of its peculiarities and genetic characteristics of its natural resources. Therefore, this project was discribed and conducted. In this investigation, cuts 2 or 3 cm of fin tissue of  specimen of Cobia obtained from Sistan and Bluchestan, Hormozgan, Bushehr and Khuzestan water provinces, were collected. DNA was extracted by Phenol-chlorophorm method and produced PCR product in length of 1060 and 1450 base pair of two mitochondrial genes COI and NADH2. Using 13 cutting enzymes (4 enzymes were subscriber for both of genes), 205 base pair (from 2510 base pair, equal with %3.8 from gene regains) were directly investigated. But binding patterns of enzymatic digestion of PCR products of both COI and ND genes from electrophoresis were monomorph in all samples and no polymorphism was observed. This may be attributed to the unsuitable choice of COI and ND2 genes for showing of intra specific divergence. But in general non-existence of genetic diversity or noticeable decrease of that among individuals has been reported in regions were fish migration exist and they can freely move between two regions. Therefore, non-observation of polymorphism in the study area might be the case and indicates represents the area. On the other hand, some scientists believe that the distributions of populations in different regions are greatly affected by environmental and physical and ecological factors. Althoug Cobia is a migratory fish, but with regard to the fact that the environmental conditions are different (specially temperature and salinity) between east and west of Persian Gulf and Oman sea, there is a possibility that different genetic groups of this species exist in the regions. Of course It is clear that using more samples and enzymes from other genetically regions could produce better results. Since none of the two investigated genes didn’t show genetic divergence or polymorphism amongst the individuals of one region or between different regions, therefore, statistic analysis for estimating of haplotype diversity or nucleotide diversity and drawing of relationship tree among individuals using available softwares was not possible.

中国鸟类的DNA分类及系统发育研究概述

鸟类分类是鸟类学其他研究领域的基础,近年来分子技术的发展,以及计算机技术的应用为鸟类分类学和鸟类系统演化研究提供了新的研究手段,给传统的系统分类研究带来了新的机遇.Tautz等于2002年首先提出运用DNA序列作为生物分类系统的主要平台,即DNA分类学(DNA Taxonomy).而Hebert等于2003年则首次提出了DNA条形码(DNA Barcoding)的概念,并对其物种分类和鉴定意义予以肯定,建议利用线粒体细胞色素C氧化酶亚单位Ⅰ(COI)的特定区段来做DNA条形编码的基础.在鸟类DNA分类方面,国内学者应用线粒体基因Cut b,COI,c-mos,c-myc,12s rRNA,16s rRNA,ND2,ND3,CR,RAG-1以及核基因myoglobin introⅡ等不同片段对很多类群进行了分类探讨和系统发育研究.但是主要集中在鸡形目及雀形目鸟类.中国是鸟类多样性极其丰富的国家,近年来很多亚种、种及以上分类阶元依然存在问题,因此,中国鸟类物种的分类地位、系统发育与演化关系等依然有很多问题等待深入研究.目前国内基于COI的鸟类分类及系统发育研究有了一些报道,但是真正的DNA条形码工作尚需继续、深入地开展.

Phylogenetic relationships of Danio within the order cypriniformes: A framework for comparative and evolutionary studies of a model species

The evolutionary relationships of species of Danio and the monophyly and phylogenetic placement of the genus within the family Cyprinidae and subfamily Rasborinae provide fundamentally important phyloinformatics necessary for direct evaluations of an array of pertinent questions in modern comparative biology. Although the genus Danio is not one of the most diverse within the family, Danio rerio is one of the most important model species in biology. Many investigations have used this species or presumed close relatives to address specific questions that have lasting impact on the hypothesis and theory of development in vertebrates. Largely lacking from this approach has been a holistic picture of the exact phylogenetic or evolutionary relationships of this species and its close relatives. One thing that has been learned over the previous century is that many organismal attributes (e.g., developmental pathways, ecologies, behaviors, speciation) are historically constrained and their origins and functions are best explained via a phylogenetic approach. Herein, we provide a molecular evaluation of the phylogenetic placement of the model species Danio rerio within the genus Danio and among hypothesized closely related species and genera. Our analysis is derived from data using two nuclear genes (RAG1, rhodopsin) and five mitochondrial genes (ND4, ND4L, ND5, COI, cyt b) evaluated using parsimony, maximum likelihood, and Bayesian analyses. The family Cyprinidae is resolved as monophyletic but the subfamily Rasborinae (priority over Danioinae) is an unnatural assemblage. Danio is identified as a monophyletic group sister to a clade inclusive of the genera Chela, Microrasbora, Devario, and Inlecypris, not Devario nor Esomus as hypothesized in previous studies. Danio rerio is sister to D. kyathit among the species of Danio evaluated in this analysis. Microrasbora and Rasbora are non-monophyletic assemblages; however, Boraras is monophyletic.

A preliminary phylogenetic analysis of Metapenaeopsis (Decapoda : Penaeidae) based on mitochondrial dna sequences of selected species from the Indo-West Pacific

Phylogenetic relationships within Metapenaeopsis remain largely unknown. The modern revision of the genus suggests that the shape of the petasma, followed by the presence of a stidulating organ, are the most important distinguishing taxonomic features. In the present study, phylogenetic relationships were studied among seven Metapenaeopsis species from the Indo-West Pacific based on partial sequences of mitochondrial 16S rRNA and cytochrome c oxidase I (COI) genes. Mean sequence divergence was 6.4% for 16S and 15.8% for COI. A strikingly large nucleotide distance (10.0% for 16S and 16.9% for COI) was recorded between M. commensalis, the only Indo-West Pacific species with a one-valved petasma, and the other species with a two-valved petasma. Phylogenetic analyses using neighbor-joining, maximum parsimony, and maximum likelihood generated mostly identical tree topologies in which M. commensalis is distantly related to the other species. Two clades were resolved for the remaining species, one with and the other without a stridulating organ, supporting the main groupings of the recent taxonomic revision. Results of the present study also indicate that the deep-water forms represent a relatively recent radiation in Metapenaeopsis.

Motilin/ghrelin荷尔蒙基因家族及其受体基因家族的分子进化研究和云南闭壳龟（Cuorayunnanensis）的分子鉴定及进化地位研究

哺乳动物motilin/ghrelin荷尔蒙基因家族编码产生三种多肽荷尔蒙，motilin、ghrelin和obestatin。这三种荷尔蒙分别结合各自特异性受体MLNR、GHSR和GPR39，调控不尽相同，但类似或相关的生理生化过程，并且它们的受体相互之间也高度同源。根据达尔文的进化论，任何复杂的生物系统都是在原有基础上，在自然选择的作用下逐步调整优化而来（Darwin 1859）。一方面，一个系统中相互作用的各个组分在进化过程中同时发生或变化的概率微乎其微；另一方面，紧密相互关联的系统中一个组分的孤立存在看起来又是毫无意义的。本研究中，我们基于系统发育分析重建了荷尔蒙基因家族及其受体基因家族的进化历史，探讨它们在进化中的关系。从而了解在一个整合的系统中，基因复制（gene duplication）后产生的新的组分是如何演化的，是如何形成新的分子间相互作用（如荷尔蒙–受体间特异性相互作用）和发生功能分化的。 我们的研究结果表明，preproghrelin（GHRL）和prepromotilin（MLN）源于一个祖先基因，是由基因重复而来，基因重复发生在C端两个新的翻译后剪辑位点演化出现之后，既两栖动物与羊膜动物分歧之后（而后产生了新的多肽荷尔蒙，例如高等哺乳动物GHRL中的obestatin）。受体与配体的进化历史并不一致。受体GPR39最先分歧，然后类似GHSR的祖先基因经历先后两次重复，产生了硬骨鱼世袭特异的基因簇A，MLNR和GHSR，基因重复事件的发生早于硬骨鱼与四足动物的分歧。Ghrelin/GHSR信号通路系统从硬骨鱼到哺乳动物的进化过程中十分保守，结构和功能几乎都没有发生太大变化。Motilin和MLNR间的特异性相互作用是荷尔蒙基因重复发生后，配体、受体间协同进化的结果，自此形成了新的Motilin/MLNR信号通路系统。 我们提出了一个复杂系统（分子间相互作用网络）进化的模型：基因重复或酶饰作用后产生了新的组分，它们通常是先前已有组分的一种结构上的变异，能与之前已经存在的分子形成新的分子间相互作用，从而演化出新的功能。基因重复之前，通常存在基因共享，即一种分子参与到多个过程、多种分子间相互作用。基因重复或酶饰作用生成了新组分，新的分子间相互作用，为功能上的专化和特化提供了条件。 此外我们还对新近发现的活的云南闭壳龟进行了分子鉴定，并探讨了云南闭壳龟的系统发育地位。云南闭壳龟（Cuora yunnanensis，Boulenger，1906）曾被认为已经灭绝，在保护生物学上受到广泛的关注。我们测定了三只活的云南闭壳龟线粒体COI和ND4及His、Ser、Leu tRNA序列片断，共1725碱基序列。结合闭壳龟属其它物种序列，包括之前测定的一只云南闭壳龟标本（MNHN 1907.10）的DNA序列，进行了分子系统学分析。与100年前的标本比较，无论是形态上、还是本文的分子系统结果都显示，新发现的活的云南闭壳龟确实是云南闭壳龟。同时，我们的结果确证了标本序列的可信性，揭示云南闭壳龟不是近期杂交形成的，代表了进化上独立的遗传谱系，且种内仍存在一定的遗传多样度。本文是分子系统学在濒危物种保护应用中的成功案例。我们的结果为推翻云南闭壳龟已经灭绝的观点提供了进一步强有力的分子生物学证据，但该物种极其稀少的状况提示其前景不容乐观，必须尽快采取有力的措施予以重点保护。

果蝇 virilis section 的起源和系统发育研究

果蝇 virilis section(Hsu 1949)自建立以来其合理性在果蝇系统发育中还没有得到 检验。本文以线粒体烟酰胺腺嘌呤二核苷酸脱氢酶第二亚单位基因(ND2)全序列和细 胞色素氧化酶I 基因(COI)部分片段以及核中乙醇脱氢酶基因Adh 的编码区为遗传标 记，对virilis section 内61 个物种(13 个未发表新种),共计117 个个体进行了序列测 定。通过对单个基因和合并数据集进行简约分析，以及对合并数据集进行邻接法分析 和贝叶斯分析，本研究试图对以下问题进行探讨：(1)果蝇virilis section 的合理性。 (2)各种组间的系统关系。(3)种组内部各物种间的系统关系。(4)果蝇virilis section 的进化历史。现将主要结果总结如下： 1)分子系统学研究显示果蝇virilis section 为一个紧密相关的进化簇，为果蝇virilis section 的合理性从分子遗传学的角度提供了证据。 2)系统发育分析支持果蝇polychaeta 种组为单系群，该种组与未归类物种D. fluvialis 具有较近的亲缘关系。它们形成virilis section 中早期分化出的谱系。robusta 种组、melanica 种组、quadrisetata 种组以及新种组间关系较近。其中，robusta 种亚 组和melanica 种组关系较近；quadrisetata 种组和新种组形成姐妹群，它们形成的谱 系与robusta 种组内的okadai 种亚组关系紧密。 3) polychaeta 种组内部分为两枝，姐妹种D. polychaeta 和D. asper 组成其中一枝， 杂交实验显示它们具有非对称的交配前生殖隔离。另一枝中D. latifshahi 和D. daruma 显示了较近的亲缘关系。非洲物种D. hirtipes 与采自西双版纳的D. polychaeta X 关系 较近，提示它们可能有共同的起源。 4)合并数据集支持D. angor 种组为单系群，其中D. angor A 和D. velox 的姐妹种 关系得到较高的支持。但是该种组内部的其它物种间关系还需要进一步研究。 5) 分子系统发育分析结果清楚地表明robusta 种组属于多系发生。该种组被分为 三个种亚组：okadai 种亚组，robusta 种亚组和lacertosa 种亚组。在lacertosa 种亚组 内，D. bai 与其它成员相对远缘。okadai 种亚组的物种由于具有2n = 12 条染色体，且X 染色体为棒状，因而被认为是robusta 种组中较早分化出来的类群。但是我们的 分子数据并不支持这种观点。D. moriwakii 最初被描述为robusta 种组物种，后被订正 至melanica 种组。我们的系统发育分析显示该种与robusta 种亚组具有较近的关系， 而与melanica 种组物种的关系相对较远。除D. moriwakii 外，melanica 种组新旧大陆 物种各自形成单系。在系统树上，中国品系D. tsigana 与D. longiserrata 关系较近， 而与日本品系D. tsigana 的关系较远。杂交实验结果显示D. tsigana 中国品系和日本 品系间存在不对称的交配倾向，表明该种不同的地理群体间可能正处于分化阶段，而 中国品系则有可能已经演化为不同的物种。 6)果蝇quadrisetata 种组为单系发生，其内部分为两个明显的亚世系。第一个亚 世系包括D. sp T，D. barutani，D. potamophila 和D. spIZU；另一个亚世系包括D. beppui，D. karakasa，D. quadrisetata，D. multidentata，D. perlucida 和D. pilosa，其 中D. beppui 为该谱系中最早分化出的物种。 通过估计谱系间的分歧时间，本文推测果蝇virilis section 的祖先大约于中新世早 期起源于热带地区，virilis section 内许多物种可能栖息在旧大陆的低纬度地区，然后 通过适应性辐射扩散到各地。几乎所有的新大陆virilis section 物种是旧大陆物种通过 白令陆桥迁移到新大陆而演化形成的。