Estimating divergence times among subfamilies in Nymphalidae

The mitochondrial cytochrome oxidase subunit Ⅰ （COⅠ） gene and the nuclear elongation factor 1α （EF-1α） gene were sequenced from 13 species of Nymphalidae. Phylogenetic trees of Nymphalidae, which is the largest family in butterflies, were constructed based on the sequences determined from 13 species sequenced in our laboratory and an additional 43 species obtained from GenBank using the maximum likelihood （ML） and Bayesian methods. Relative-rate tests between lineages in these phylogenetic trees were performed. On the basis of the results of the relative-rate tests and fossil information of Satyrinae, Nymphalinae and Biblidinae, the average divergence times among the subfamilies are estimated as 44.2-87.1 million years ago （Ma）. These results will be helpful for better understanding of the origin and evolution of this family, as well as the divergence time of butterflies and other complex taxa.     

Phylogenetic position of three Condylostoma species （Protozoa,Ciliophora, Heterotrichea） inferred from the small subunit rRNA gene sequence

The systematically poorly known ciliate genus Condylostoma was erected by Vincent in 1826. About 10 morphotypes have been reported, but any molecular investigations concerning this group so far are lacking. In this work, the small subunit ribosomal RNA （SS rRNA） gene of three marine Condylostoma species was sequenced, by which the phylogenetic trees were constructed by distancematrix, maximum parsimony and Bayesian inference methods. The results show that （1） all the trees have similar topologies with high supports; （2） Condylostoma is mostly related to the genus Condylostentor; and （3） three Condylostoma species as well as Condylostentor auriculatus cluster together and form a sister group with other heterotrichs. This is moderately consistent with the assessment of phylogenetic relationships of Condylostoma-related heterotrichs from the morphological information. The phylogenetic relationship of some other related heterotrichs, Peritromus, Folliculina, Stentor and Blepharisma, has been also discussed.     

Investisations into the perplexing interrelationship of the Genus Takifugu Abe, 1949 （Tetraodontiformes,Tetraodontidae）

The phylogenetic relationships within the genus Takifugu Abe, 1949 （Tetraodontiformes, Tetraodontidae） remain unresolved. Because of the use of Takifugu as model organisms, the resolution of these relationships is crucial for the interpretation of evolutionary trends in biology. Pufferfishes of this genus are comprised of a comparatively small number of species and are mainly distributed along the coastal region of the western part of the Sea of Japan and the coastline of China. Mitochondrial gene sequences were employed to test the phylogenetic hypotheses within the genus. Seventeen species of the genus were examined. Molecular phylogenetic trees were constructed using the maximum parsi- mony, neighbor-joining, maximum likelihood and Bayesian methods. Our hypothesis of internal relationships within the genus differs from previous hypotheses. Our results indicate that （1） the genus Takifugu is a monophyletic assemblage; （2） the genus is divided into 6 subgroups based on the moecular data; and （3） there is low genetic diversity among the species within this genus. In addition, speciation within Takifugu appears to be driven by hybridization and isolation by distribution. Our results also suggested that the taxonomy in the genus should be clarified based on both molecular and morphological data.     

Phylogenetic position of Ephedra rhytidosperma, a species endemic to China： Evidence from chloroplast and ribosomal DNA sequences

The chloroplast genes matK and rbcL, ribosomal gene 18S and ITS regions of nuclear ribosomal DNA from Ephedra rhytidosperma, a species endemic to China, were sequenced and its phylogenetic position was investigated. Independent and combined phylogenetic analyses for the DNA sequences from 16 taxa representing 15 species of the genus Ephedra were performed using the maximum parsimony （MP）, neighbor-joining （N J）, minimum evolution （ME） and maximum likelihood （ML） methods. The results indicate that E. rhytidosperma is closely related to E. equisetina. The divergence time between them is estimated to be 10.85±2.44 Ma based on the results of the relative-rate tests and the evolutionary rate of rbcL gene.     

The phylogenetic relationship of the family Lutjanidae based on analyses of AFLP and mitochondrial 12S rRNA sequences

Fishes of the family Lutjanidae are commercially important in South China Sea. However, the phylogeny of Lutjanids is still unclear and there are many controversies over it. Herein, studies about the phylogeny of Lutjanids were performed based on Amplified Fragment Length Polymorphism （AFLP） analysis of genome DNA and sequence analysis of mitochondrial 12S rRNA gene, and 10 Lutjanidae species and 1 Lethrinidae species were employed. The topologies of minimum evolution （ME） trees based on the two analyses respectively were congruent except for positions of genera Pristipomoides and Caesio. The optimal substitution model TrN ＋ G for DNA sequences of 12S rRNA genes in Lutjanids was obtained using MODELTEST 3.6 software and maximum likelihood （ML） analysis supports the topology displayed by the ME tree. The test of log-likelihood suggests that the use of molecular clock calibrations to estimate species divergence time appeared valid. Phylogenetic analyses using AFLP data and DNA sequences of mitochondrial 12S rRNA genes indicated the monophyly of Lutjanus genra. However, further studies are required to reveal the phylogenetic relationship among other genera. In addition, the results demonstrated that AFLP genetic marker was suitable for the phylogenetic analysis of Lutjanids.     

Asian origin for Polystichum （Dryopteridaceae） based on rbcL sequences

Chloroplast rbcL sequences of 60 species of Polystichum sensu lato (s.l.), including 23 new sequences from southwest China, were used to assess the phylogenetic relationships within the genus. On the basis of estimated evolution rate of rbcL gene and the genetic distance data that passed relative-rate tests, we further estimated the divergence times between some clades of the genus. The phylogenetic relationships were inferred using the neighbor-joining and maximum-parsimony methods, both methods producing trees with completely congruent topology. These trees reveal that all species of Polystichum s.l. in this study (including Cyrtomium and Cyrtomidictyum) form a monophyletic group. The basal split in Polystichum s.l. separates a clade with all Asian members from a clade containing other species from all over the world. The phylogenetic and divergence time estimation results lead us to suggest that Polystichum s.l. originated in Asia in the late Late Cretacous (≈76 Ma) and migrated into other places in the world in early Eocene(≈46 Ma).     

Phylogenetic relationships of host insects of Cordyceps sinensis inferred from mitochondrial Cytochrome b sequences

This study used the sequence of the mitochondrial Cytochrome b(Cytb)to estimate phylogenetic relationships among host Hepialidae insects of Cordyceps sinensis.Genome DNA of host insect was extracted from the dead larva head part of 18 cordyceps populations and 2 species of Hepialus,and the Cytb fragment of host insect was amplified with PCR technique.The nucleotide sequence alignments and their homologous sequences of 24 species host Hepialidae insects of Cordyceps sinensis were obtained from GenBank and were used to construct phylogenetic trees based on neighbor-joining method.The results showed that genus Bipectilus diverged earlier than genus Hepialus and Hepialiscus.Hepialus host insects of Cordyceps sinensis have multitudinous species with different morphological characteristics and geographical distributions.The interspecific genetic differentiations are obvious in Hepialus.Thus,the genus Hepialus might be considered as polyphyletic origin.Cytb sequences have abundant variations among the host insects of Cordyceps sinensis on spe- cific and generic level.The divergence rate of Cytb sequences among the species in Hepialus ranged from 0.23% to 9.24%,except that Hepialus pratensis and Hepialus jinshaensis have the same sequence.Cytb sequence can be used for species identification of host insects of Cordyceps sinensis,but further confirmation in more host insect species is needed.To obtain the Cytb sequence of host insect by ampli- fying DNA extracted from the head part of dead larva in cordyceps turns out to be an effective and accurate approach,which will be useful for studies on phylogeny and genetic structure of host insects of cordyceps populations,especially for analyzing relationships between C. sinensis and its host insects.     

Investigations into the perplexing interrelationship of the Genus Takifugu Abe, 1949 (Tetraodontiformes, Tetraodontidae)

The phylogenetic relationships within the genus Takifugu Abe, 1949 (Tetraodontiformes, Tetraodonti- dae) remain unresolved. Because of the use of Takifugu as model organisms, the resolution of these relationships is crucial for the interpretation of evolutionary trends in biology. Pufferfishes of this ge- nus are comprised of a comparatively small number of species and are mainly distributed along the coastal region of the western part of the Sea of Japan and the coastline of China. Mitochondrial gene sequences were employed to test the phylogenetic hypotheses within the genus. Seventeen species of the genus were examined. Molecular phylogenetic trees were constructed using the maximum parsi- mony, neighbor-joining, maximum likelihood and Bayesian methods. Our hypothesis of internal relationships within the genus differs from previous hypotheses. Our results indicate that (1) the genus Takifugu is a monophyletic assemblage; (2) the genus is divided into 6 subgroups based on the mo- lecular data; and (3) there is low genetic diversity among the species within this genus. In addition, speciation within Takifugu appears to be driven by hybridization and isolation by distribution. Our re- sults also suggested that the taxonomy in the genus should be clarified based on both molecular and morphological data.     

Phylogenetic studies of the core Alismatales inferred from morphology and rbcL sequences

The phylogeny of Alismatales remains an area of deep uncertainty, with different arrangements being found in studies that examined various subsets of genes and taxa. Herein we conducted separate and combined analyses of 103 morphological characters and 52 rbcL sequences to explore the controversial phylogenies of the families. Congruence between the two data sets was explored by computing several indices. Morphological data sets contain poor phylogenetic signals. The homology of morphological characters was tested based on the total evidence of phylogeny. The incongruence between DNA and morphological results; the hypothesis of the ‘Cymodoceaceae complex’; the relationships between Najadaceae and Hydrocharitaceae; the intergeneric relationships of Hydrocharitaceae; and the evolutionary convergence of morphological characters were analyzed and discussed.     

Phylogenetic relationships and divergence times of the family Araucariaceae based on the DNA sequences of eight genes

Araucariaceae is one of the most primitive families of the living conifers, and its phylogenetic relationships and divergence times are critically important issues. The DNA sequences of 8 genes, i.e., nuclear ribosomal 18S and 26S rRNA, chloroplast 16S rRNA, rbcL, matK and rps4, and mitochondrial coxl and atpl, obtained from this study and GenBank were used for constructing the molecular phylogenetic trees of Araucariaceae, indicating that the phyiogenetic relationships among the three genera of this family should be （（Wollemia, Agathis）, Araucaria）. On the basis of the fossil calibrations of Wollemia and the two tribes Araucaria and Eutacta of the genus Araucaria, the divergence time of Araucariaceae was estimated to be （308± 53） million years ago, that is, the origin of the family was in the Late Carboniferous rather than Triassic as a traditional view. With the same gene combination, the diver- gence times of the genera Araucaria and Agathis were （246 ± 47） and （61 ±15） Ma, respectively. Statis- tical analyses on the phylogenetic trees generated by using different genes and comparisons of the divergence times estimated by using those genes suggested that the chloroplast matK and rps4 genes are most suitable for investigating the phylogenetic relationships and divergence times of the family Araucariaceae.     

鲟形目线粒体16S rRNA的部分序列变异及系统发生

采用通用引物对3种鲟形目（Acipenseriformes）鱼类,中华鲟（Acipenser sinensis）、俄罗斯鲟（Acipenser gueldenstaedtii）和匙吻鲟（Polyodon spathula）的16S rRNA基因部分片段进行扩增,并与NCBI数据库中获得另外6种鲟形目鱼类相应的16S rRNA序列部分进行比较,探讨鲟形目2个科5个属共9个种之间的遗传变异和亲缘关系.数据合并后用于分析的序列共514 bp,变异位点29个,含简约信息位点16个,单一信息位点13个.以多鳍鱼目（Polypteriformes）多鳍鱼属Polypterus ornatipinnis为外群,采用部分16S rRNA序列基于Kimura双参数模型构建其系统发育关系,结果表明：MP法和NJ法得到系统树基本相同,其科、亚科和属的分化上基本与传统形态学分类相吻合,另外,欧鳇与鲟属鱼类的分化也并不明显.值得注意的是,鲟亚科6个种所划分出的3个分支恰恰分别归属于3个地理区域,欧鳇、闪光鲟和俄罗斯鲟主要集中于黑海和里海流域等地区;中华鲟和达氏鲟分布在西北太平洋,中国长江流域以及朝鲜;而高首鲟则分布于东太平洋.     

位点特异性频率模型在昆虫分子系统发育分析中的应用

根据位点特异性频率模型(site-specific rate model),利用PAUP软件,对线粒体COI基因序列不同密码子位点的数据模块分别进行了豉甲科和水生肉食亚目在亚科或科水平上的系统发育学分析,结果表明第二密码子数据模块获得的系统分析结果与形态学分析结果一致。表明位点特异性频率模型在线粒体COI基因作为标记来进行昆虫分子系统发育分析上获得了更好的效果。     

基于12S、16S和Cyt b基因部分序列的丽棘蜥系统发育分析

本文测定丽棘蜥12SRNA、16SRNA、Cyt 6基因部分序列,并从GenBank中下载15种有鳞目动物的同源序列,以鳄目的2个种(Alligator sinensis和Cairman crocodilus)作为外群,构建MP、NJ和ML分子系统树,探讨它们的系统发育关系.MP、NJ和ML分子系统树聚类的结果显示:蛇蜥亚目、壁虎亚目、蚓蜥亚目、石龙子亚目和鬣蜥亚目能够完全分开,蛇亚目和蜥蜴亚目在有鳞目中为姐妹支关系.     

基于16S rRNA序列的鳃金龟亚科5族种类的系统发育关系

测定了鳃金龟亚科部分种类的线粒体16SrRNA部分序列,运用MEGA 4.0、PAUP*4.0b10和MrBayes等软件,对5个族29个代表种的序列变异和系统关系进行了研究.序列变异分析结果显示:绢金龟族Sericini、哦鳃金龟族Hopliini、鳃金龟族Melolonthini的族内遗传距离分别为7.7%,11.3%,10.6%,族间遗传距离在12.6%～19.2%.最大似然树(ML)和贝叶斯树(BI)结果表明,所有种类分别聚集在所属的族下,哦鳃金龟族、绢金龟族分别为单系.齿爪鳃金龟属Holotrichia、云鳃金龟属Polyphylla、胸突鳃金龟属Hoplosternus、Hilyotrogus、双绺鳃金龟属Amphimallon、Hoplochelus、婆鳃金龟属Brahmina和皱鳃金龟属Trematodes聚在鳃金龟族分支下,没有形成明显的根鳃金龟族Rhizotrogini分支.齿爪鳃金龟属的非单系性与前人研究结果一致.皱鳃金龟属Trematodes和婆鳃金龟属Brahmina的代表种为姐妹种.绢金龟族的绢金龟属Serica和玛绢金龟属Maladera多数种类聚在本属分支下.证明16SrRNA序列可探讨鳃金龟亚科高级阶元的系统发育关系.     

基于叶绿体rbcL等3个基因的木灵藓科植物属的系统发育

以木灵藓科21属33种植物为内类群,以缩叶藓科（Ptycomitriaceae）、虎尾藓科（Hedwigiaceae）、树生藓科（Erpodiaceae）、蕨藓科（Pterobryaceae）、平藓科（Neckeriaceae）、蔓藓科（Meteoriaceae）、刺果藓科（Symphyodontaceae）、油藓科（Hookeriaceae）、灰藓科（Hypnaceae）和金发藓科（Poltrichaceae）为外类群,以NADH脱氢酶亚基〖STBX〗5〖STB3〗基因、叶绿体rbcL基因和tRNA-Leu基因为分析指标,从Genbank获得它们的碱基序列,通过Clustalx对碱基比对,使用Winclada 软件对木灵藓科植物进行了分支系统学分析.结果显示: (1) 木灵藓科是一个明确的单系类群;(2) 支持将木灵藓科分为两个亚科: 木灵藓亚科（Orthotrichoideae）和蓑藓亚科（Macromitrioideae）;(3) 马他属（Bryomaltaea）归为蓑藓亚科为宜.     

基于trnL-F基因序列的铁兰属植物亲缘关系分析

以莺哥凤梨(Vriesea carinata)为外类群,利用Clustal X1.81和MEGA3.1软件对31种铁兰属植物的基因序列进行比对和分析,并构建分子系统树。结果表明:31种铁兰属植物供试材料的trnL-F序列长度在806～835 bp之间,变异位点22个,信息位点14个,平均遗传距离0.007,且在301～322 bp和826～830 bp处分别出现了插入和polyT序列。利用ML法建立的系统树显示,31个铁兰属植物分为3组。序列分析结果与形态学结果基本一致,表明trnL-F序列可用于铁兰属植物的亲缘关系分析,并可作为某些特定种的鉴定依据。     

DNA条形码技术在白洋淀流域浮游动物调查中的应用

DNA 条形码在物种鉴定中得到了有效的应用,有助于分类学和生物多样性研究. 本研究基于线粒体COI基因序列,使用邻接法(neighbor joining method,NJ)和最大似然法(maximum Likelihood,ML)构建系统发育树,对2018年5月在白洋淀流域采集的浮游动物样品进行初步研究. 结果表明,在对桡足类的系统发育树分析时,NJ树和ML树的结果保持一致,而且和形态学鉴定结果保持一致. 在对枝角类的系统发育树的分析中,NJ树和ML树的结果有所差异,和形态学鉴定结果也有所不同,分析原因,可能是由于拼接序列时剪短了部分序列,使序列间差异性变小引起误差. 总体上看,基于COI基因的DNA条形码可以有效地应用到白洋淀流域的浮游动物鉴定中,并可以逐步完善,最终建立一个完整的DNA条形码数据库.     

中国斑腿蝗科线粒体基因的分子系统学研究(英文)

使用线粒体基因COⅠ,COⅡ,Cytb分析并建立中国斑腿蝗科部分种属的系统发育关系,其中对7个新种(Apalacris eminifronta,Caryanda pseudodentata,Caryanda bannaensis,Caryanda ruiliensis,Longchuanacris fui,Longchuanacris xiaoheis-hanensis,Podismodes dabieshanensis)的归属从分子角度也做了讨论。本研究将实验所获得的56条序列与NCBI中所收录的斑腿蝗科序列进行联合分析,采用最大似然法、贝叶斯推论等系统分析方法并综合P距离进行分析,最终得出以下几点结论:①龙川蝗属Longchuanaceis Zheng et Fu划归卵翅蝗亚科Caryandinae;②蹦蝗属Sinopodisma Chang、云秃蝗属Yunnanacris Chang均为华秃蝗属Podismodes Ramme的同物异名;③不支持Storozhenko将豫蝗属Yupodisma Zhang et Xia并入安秃蝗属AnapodismaDovnar-Zapolskii,从本研究所建的系统发育树来看,豫蝗属仍应作为秃蝗亚科Podisminae的一个独立属;④前人利用形态特征鉴定的七个新种与分子系统发育分析结果一致,新种独立有效。     

9种拟步甲16S rDNA部分序列及其亲缘关系

测定了9种拟步甲的16S rDNA部分基因序列,并与GenBank中的1种步甲的基因序列作同源性比较,计算其核苷酸使用频率并构建了分子系统树.在获得的435 bp的序列中,A+T约占74.4%,颠换(transversion)取代的速率大于转换(transition)取代的速率,其中277个核苷酸位点存在变异.结果表明:属内种间的碱基序列差异范围为3.4%～6.2%;族内属间为9.4%～11.0%;科内族间为10.8%～17.7%;科与科间的差异达到46.7%～50.3%.分子系统树表明:拟步甲科为一单系群,其中琵甲族较为进化,漠甲族与漠王族相对原始;琵甲族与土甲族的亲缘关系较近;漠甲族、漠王族与鳖甲族的亲缘关系较近.本结果与传统的分类观点相吻合.