Molecular Phylogeny and Species Identification of Pufferfish of the Genus Takifugu (Tetraodontiformes, Tetraodontidae)

The phylogenetic relationships and species identification of pufferfishes of the genus Takifugu were examined by use of randomly amplified polymorphic DNA (RAPD) and sequencing of the amplified partial mitochondrial 16S ribosomal RNA genes. Amplifications with 200 ten-base primers under predetermined optimal reaction conditions yielded 1962 reproducible amplified fragments ranging from 200 to 3000 bp. Genetic distances between 5 species of Takifugu and Lagocephalus spadiceus as the outgroup were calculated from the presence or absence of the amplified fragments. Approximately 572 bp of the 16S ribosomal RNA gene was amplified, using universal primers, and used to determine the genetic distance values. Topological phylogenic trees for the 5 species of Takifugu and outgroup were generated from neighbor-joining analysis based on the data set of RAPD analysis and sequences of mitochondrial 16S rDNA. The genetic distance between Takifugu rubripes and Takifugu pseudommus was almost the same as that between individuals within each species, but much smaller than that between T. rubripes, T. pseudommus, and the other species. The molecular data gathered from both analysis of mitochondria and nuclear DNA strongly indicated that T. rubripes and T. pseudommus should be regarded as the same species. A fragment of approximately 900 bp was amplified from the genome of all 26 T. pseudommus individuals examined and 4 individuals of intermediate varieties between T. rubripes and T. pseudommus. Of the 32 T. rubripes individuals, only 3 had the amplified fragment. These results suggest that this fragment may be useful in distinguishing between T. rubripes and T. pseudommus. Received September 29, 2000; accepted February 26, 2001.     

Molecular Phylogeny Analysis and Species Identification of Dendrobium (Orchidaceae) in China

Dendrobium plants are important commercial herbs in China, widely used in traditional medicine and ornamental horticulture. In this study, sequence-related amplified polymorphism (SRAP) markers were applied to molecular phylogeny analysis and species identification of 31 Chinese Dendrobium species. Fourteen SRAP primer pairs produced 727 loci, 97% of which (706) showed polymorphism. Average polymorphism information content of the SRAP pairs was 0.987 (0.982–0.991), showing that plenty of genetic diversity exists at the interspecies level of Chinese Dendrobium. The molecular phylogeny analysis (UPGMA) grouped the 31 Dendrobium species into six clusters. We obtained 18 species-specific markers, which can be used to identify 10 of the 31 species. Our results indicate the SRAP marker system is informative and would facilitate further application in germplasm appraisal, evolution, and genetic diversity studies in the genus Dendrobium.     

Molecular identifications uncover diverse fungal symbionts of Pleione (Orchidaceae)

Fungal mutualisms are essential for the evolution and diversification of Orchidaceae, yet the fungal symbionts of Pleione orchids are poorly understood because molecular data are unavailable for this genus. Based on ITS-rDNA sequencing for mycobionts of 15 Pleione species (both wild and cultivated plants were included), we conducted phylogenetic analyses for the most dominant mycobionts, and compared the operational taxonomic units (OTUs) of mycorrhizal fungi among species within Pleione. Tulasnellaceae, Ceratobasidiaceae, Serendipitaceae (Sebacinales), Atractiellales, and Auriculariales were reported as putative mycobionts of Pleione. In particular, the mycorrhizal associations between subtropical orchids and Atractiellales have not been observed before. For the dominant mycobionts in the roots of Pleione and its related genera, Bletilla and Coelogyne, we detected no fungal OTU that was shared. Within Pleione, species with a sympatric distribution showed preferences for different fungi. Epiphytic and lithophytic individuals of Pleione albiflora shared OTUs of Tulasnellaceae but harbored different OTUs of Sebacinales, indicating some degree of fungal specificity toward certain habitats. These findings provide new insights into the ecological adaptation and evolution of orchids, and will contribute to the conservation and utilization of species resources.     

基于孢子形态和分子证据探讨鳞盖蕨属(碗蕨科)系统分类

孢粉学是解决植物分类中疑难类群物种微形态分化的重要方法, 随着分子系统学的发展, 结合这两门学科的优势可以更加有效地解决疑难类群的分类学问题。鳞盖蕨属(Microlepia)是一个分类困难的疑难类群, 采用孢粉学与分子系统学一一对应的方法, 以及居群取样方式, 选取280份样本, 联合4个叶绿体片段(rbcL、trnL-F、psbA-trnH和rps4), 采用最大似然法和贝叶斯法构建该属的系统发生关系, 在此基础上对凭证标本中100份材料的孢子进行观察和分析。综合分子系统学和孢粉学的研究结果, 得出结论: (1) 在形态学研究中广泛被接受的15个物种得到了单系支持, 并厘清了分类困难的复合群; (2) 发现边缘鳞盖蕨(M. marginata)可能存在隐性种; (3) 建议恢复过去归并处理为异名的瑶山鳞盖蕨(M. yaoshanica)、罗浮鳞盖蕨(M. lofoushanensis)、四川鳞盖蕨(M. szechuanica)以及滇西鳞盖蕨(M. subspeluncae); (4) 提出鳞盖蕨属可能存在杂交现象; (5) 提出鳞盖蕨属完整的属下分类建议。     

中国兰科石豆兰属一新种

描述了中国石豆兰属Bulbophyllum一新种——天贵卷瓣兰（Bulbophyllum tianguii K．Y．Lang ＆ D．Luo）,并提供墨线图。该种产于广西乐业县,与富宁卷瓣兰B．funingense Z．H．Tsi＆S．C．Chen外貌颇相似,但中萼片先端渐尖并具3mm长的芒,侧萼片宽4～4．5mm,花瓣先端渐尖并具1．5mm长的芒,唇盘具2条肉质、具缘毛的纵脊,可与后者明显区别。     

Liparis tsii: a new species of Orchidaceae (tribe-Malaxideae) from Guangdong,China with its phylogenetic position

Liparis tsii, a new orchid from Guangdong, China, is described and illustrated. It is compared with its closest allies: L. sasakii Hayata, L. nanlingensis H.Z. Tian & F.W. Xing, L. krameri Franch. & Sav., L. reckoniana T.C. Hsu and L. brunnea Ormerod belonging to section Liparis. Further phylogenetic study based on combined molecular data from nrITS and plastid trnL-F ascertained its phylogenetic position as L. tsii being sister to L. sasakii and L. nanlingensis. A key to identification with the allied species is provided.     

Ontogenesis and Molecular Phylogeny of the Marine Ciliate Diophryopsis hystrix: Implications for the Systematics of the Diophrys-like Species (Ciliophora, Spirotrichea, Euplotida)

ABSTRACT. The cortical development of Diophryopsis hystrix , a small marine euplotid ciliate, was observed during binary division. The pattern of morphogenesis is generally similar to that of other Diophrys- like species, with the exception of the marginal cirrus, which develops in a unique manner in D. hystrix being formed from the posterior end of the dorsal kinety anlage, in a similar manner to the formation of the caudal cirri. In the small subunit (SS) rRNA gene sequence tree, Diophryopsis represents a distinct clade separate from Diophrys . Furthermore, our ontogenetic and SSrRNA gene sequence data both support the suggestion that the Diophrys complex is sister to Uronychia , and should be considered as a distinct subfamily within the family Uronychiidae, i.e. Diophryinae Jankowski, 1979, comprising Diophrys , Diophryopsis , and Paradiophrys . Diagnoses of the subfamilies Diophryinae and Uronychiinae are provided.     

A phylogenetic analysis of the Orchidaceae: evidence from rbcL nucleotide

Cladistic parsimony analyses of rbcL nucleotide sequence data from 171 taxa representing nearly all tribes and subtribes of Orchidaceae are presented here. These analyses divide the family into five primary monophyletic clades: apostasioid, cypripedioid, vanilloid, orchidoid, and epidendroid orchids, arranged in that order. These clades, with the exception of the vanilloids, essentially correspond to currently recognized subfamilies. A distinct subfamily, based upon tribe Vanilleae, is supported for Vanilla and its allies. The general tree topology is, for the most part, congruent with previously published hypotheses of intrafamilial relationships; however, there is no evidence supporting the previously recognized subfamilies Spiranthoideae, Neottioideae, or Vandoideae. Subfamily Spiranthoideae is embedded within a single clade containing members of Orchidoideae and sister to tribe Diurideae. Genera representing tribe Tropideae are placed within the epidendroid clade. Most traditional subtribal units are supported within each clade, but few tribes, as currently circumscribed, are monophyletic. Although powerful in assessing monophyly of clades within the family, in this case rbcL fails to provide strong support for the interrelationships of the subfamilies (i.e., along the spine of the tree). The cladograms presented here should serve as a standard to which future morphological and molecular studies can be compared.     

海南石斛化学成分研究

为了解海南石斛(Dendrobium hainanense Rolfe)的化学成分,采用色谱技术从海南石斛茎叶中分离得到14个化合物,经波谱分析分别鉴定为:2,6-二甲氧基对苯醌(1)、(+)-dehydrovomifoliol(2)、blumenol A(3)、2,7-二羟基-3,4-二甲氧基-9,10-二氢菲(4)、2,7-二羟基-3,4-二甲氧基菲(5)、3,7-二羟基-2,4-二甲氧基菲(6)、3-羟基-2,4,7-三甲氧基-9,10-二氢菲(7)、3-羟基-2,4,7-三甲氧基菲(8)、3,4,7-三羟基-2-甲氧基菲(9)、3,7-二羟基-2,4-二甲氧基-9,10-二氢菲(10)、(+)-lyoniresinol(11)、丁香脂素(12)、denchrysan A(13)和nobilone(14)。这些化合物均为首次从海南石斛中分离得到。活性测试结果表明化合物4~6、8~9、11和14对乙酰胆碱酯酶有抑制活性。     

中国兰科石豆兰属一新种

描述了中国石豆兰属Bulbophyllum一新种——天贵卷瓣兰(Bulbophyllum tianguii K.Y.Lang & D.Luo),并提供墨线图。该种产于广西乐业县,与富宁卷瓣兰B.funingense Z.H.Tsi & S.C.Chen外貌颇相似,但中萼片先端渐尖并具3 mm长的芒,侧萼片宽4~4.5 mm,花瓣先端渐尖并具1.5 mm长的芒,唇盘具2条肉质、具缘毛的纵脊,可与后者明显区别。     

基于COI基因序列的九种真蝽属昆虫的分子系统学研究(半翅目:蝽科)

通过对真蝽属Pentatoma 9种昆虫线粒体COI基因约798bp的序列进行分子进化分析,并以同蝽科宽铗同蝽Acanthosoma labiduroides为外群,采用最大简约法、最大似然法和邻接法构建了分子系统树,来探讨真蝽属的系统发育关系.研究结果支持褐真蝽群P. semiannulata-group的划分,绿角真蝽Pentatoma viridicornuta应划归到褐真蝽群P. Semiannulata-group;红足真蝽群中的角肩真蝽P. angulata与红足真蝽P. rufipes遗传距离较小,它们是否为1个物种值得关注;真蝽属各群之间的系统发育关系以及是否可分为3个属值得进一步研究.     

Implications of Molecular and Morphological Data for Understanding Ateline Phylogeny

The ateline monkeys constitute as certain a monophyletic group as there is among primates. The group is intriguing because while their adaptations are well-documented and their monophyly as a group is unquestioned, their phylogenetic interrelationships are controversial. Molecular data indicate a phylogeny at odds with their morphology. Traditional morphological comparisons isolate Alouatta from the atelins, and link Ateles and Brachyteles as a sister group to the exclusion of Lagothrix. In contrast, several recent molecular studies point to a closer relationship between Brachyteles and Lagothrix than between Brachyteles and Ateles. At the heart of the problem lie the assumptions we make about the validity of data and the homology of observed traits. The fossil record further confounds the issue. We must account for the fossil record because it is positive evidence. But we cannot control how much of it there is or how much of it ever will be known. At this point in time, the ateline molecular and fossil record provoke us to examine critically our morphological approach to phylogenetic modeling.     

Species Diversity,Phylogeny and Large Scale Biogeographic Patterns of the Genus Padina (Phaeophyceae,Dictyotales)

The brown algal genus Padina (Dictyotales, Phaeophyceae) is distributed worldwide in tropical and temperate seas. Global species diversity and distribution ranges, however, remain largely unknown. Species‐level diversity was reassessed using DNA‐based, algorithmic species delineation techniques based on cox3 and rbcL sequence data from 221 specimens collected worldwide. This resulted in estimates ranging from 39 to 61 putative species (ESUs), depending on the technique as well as the locus. We discuss the merits, potential pitfalls, and evolutionary and biogeographic significance of algorithmic species delineation. We unveil patterns whereby ESUs are in all but one case restricted to either the Atlantic or Indo‐Pacific Ocean. Within ocean basins we find evidence for the vast majority of ESUs to be confined to a single marine realm. Exceptions, whereby ESUs span up to three realms, are located in the Indo‐Pacific Ocean. Patterns of range‐restricted species likely arise by repeated founder events and subsequent peripatric speciation, hypothesized to dominate speciation mechanisms for coastal marine organisms in the Indo‐Pacific. Using a three‐gene (cox3, psaA and rbcL), relaxed molecular clock phylogenetic analysis we estimated divergence times, providing a historical framework to interpret biogeographic patterns.     

A Molecular Phylogeny of the Lichen Genus Lecidella Focusing on Species from Mainland China

The phylogeny of Lecidella species is studied, based on a 7-locus data set using ML and Bayesian analyses. Phylogenetic relationships among 43 individuals representing 11 Lecidella species, mainly from mainland China, were included in the analyses and phenotypical characters studied and mapped onto the phylogeny. The Lecidella species fall into three major clades, which are proposed here as three informal groups–Lecidella stigmatea group, L. elaeochroma group and L. enteroleucella group, each of them strongly supported. Our phylogenetic analyses support traditional species delimitation based on morphological and chemical traits in most but not all cases. Individuals considered as belonging to the same species based on phenotypic characters were found to be paraphyletic, indicating that cryptic species might be hidden under these names (e.g. L. carpathica and L. effugiens). Potentially undescribed species were found within the phenotypically circumscribed species L. elaeochroma and L. stigmatea. Additional sampling across a broader taxonomic and geographic scale will be crucial to fully resolving the taxonomy in this cosmopolitan genus.     

A new species ofPsilochilus(Orchidaceae) from the Dominican Republic

Psilochilus crenatifolius, a new species from the Dominican Republic, is described and illustrated. It is only known from the type collection. The taxonomic affinity of the new entity is briefly discussed.     

27种瓢虫mtDNA-COI基因序列分析及系统发育研究(鞘翅目:瓢虫科)

将自测的瓢虫科4亚科16种和从GenBank中检索到相关物种的mtDNA-COI基因编码区序列片断进行同源性比较,计算核苷酸使用频率,并构建分子系统树。在获得的471bp序列中,共有227个变异位点,195个简约信息位点,A T约占66.7%,转换发生比颠换更频繁。分子系统树表明:同种和同属的瓢虫均以较高置信值聚在一起,4个亚科都能恢复为单系;食植瓢虫亚科从系统树的基部最早分出,是瓢虫科最为原始的类群,小毛瓢虫亚科和盔唇瓢虫亚科首先聚为一支,显示二者具有较近的亲缘关系,然后再与瓢虫亚科相聚,食植瓢虫亚科的食植瓢虫属和裂臀瓢虫属均不是单系群。     

Osmophore structure and phylogeny of Cirrhaea (Orchidaceae,Stanhopeinae)

Phylogenetic relationships and osmophore evolution of six Cirrhaea spp. were studied. Floral morphology was analysed using fresh flowers, and osmophore anatomy was determined on the basis of fixed flowers. Phylogenetic relationships of Cirrhaea were inferred on the basis of internal transcribed spacer (ITS), matK and trnL‐F regions using maximum parsimony and Bayesian analyses. Floral morphology and osmophore structure vary among species. All Cirrhaea osmophores have a secretory epidermis without papillae. Cirrhaea is monophyletic and includes three subclades: (1) C. dependens/C. nasuta, with a secretory cylindrical protuberance at the base of the labellar midlobe; (2) C. fuscolutea/C. longiracemosa, with a secretory tissue at the base of the shell‐shaped midlobe; and (3) C. loddigesii/C. seidelii with secretory tissue on the inner surface of the lateral lobes. The features of the flowers and osmophores in Cirrhaea spp. extend our knowledge of the diversity of secretory structures in Stanhopeinae, and demonstrate that floral morphology reflects phylogenetic relationships in Cirrhaea. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176 , 369–383.     

Molecular Phylogeny of the Genus Paramesotriton (Caudata: Salamandridae)

To elucidate the phylogeny of the genus Paramesotriton (Caudata: Salamandridae), we investigated three mitochondrial DNA gene fragments (1207 bp in total) of cytochrome b, ND2, and ND4 for its six recognized species. The phylogenetic relationships within Paramesotriton were reconstructed by maximum parsimony (MP) and maximum likelihood (ML) methods. Phylogenetic trees (MP and ML trees) that were constructed from the combined data set of the three gene fragments indicated that all six species of Paramesotriton formed a monophyletic group, with P. caudopunctatus as basal to the other five species. This result suggests that P. fuzhongensis is a valid species in Paramesotriton.