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.     

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.     

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.     

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.     

虫草属多元起源的分子生物学证据

测定了虫草属 (Cordyceps)代表种及其相关无性型菌 (anamorphicstate )核糖体rDNAITS区序列 ,并从国际分子生物学数据库中调取相关序列 ,以Phylip35分析软件构建序列距离矩阵和分子系统发育树图 ,结果表明 :虫草属各个种之间ITS区序列差别比较大 ,不能形成一个单系的类群 .冬虫夏草 ,古尼虫草 ,蛹虫草 ,大团囊虫草在系统发育上分别与丝孢纲(Hyphomycetes) ,束梗孢目 (Stilbelaes)的中国被毛孢 ,丝孢目 (Hyphomycetales)的黄绿绿僵菌 ,丝孢目 (Hyphomycetales)的球孢白僵菌 ,担子菌纲的Cyphomyrmexminutus非常接近 ,在树图上分别处于相对独立的分支 ;说明形态学上划分的虫草属并非一自然发生的类群 ,也说明了虫生真菌形态与基因水平进化的不一致性 .     

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).     

五倍子蚜与寄主植物DNA序列系统发育关系及其协同进化

五倍子蚜特指寄生在漆树科盐肤木属几种植物上形成五倍子虫瘿的一类蚜虫.本研究采用PCR直接测序方法测定了五倍子蚜10个种共19个样本的mtDNA COI基因部分DNA序列(660 bp),从GenBank下载瘿绵蚜科Pemphigidae瘿绵蚜属Pemphigus、卷叶绵蚜属Prociphilus和棉蚜属Eriosoma及纩蚜科Mindaridae纩蚜属Min-darus种相应基因序列为外类群,构建五倍子蚜种之间的MP和ML分子系统发育关系,两种方法得到一致的结果.五倍子蚜10个种共形成3个大的聚类簇,圆角倍蚜属的红倍花蚜和倍花蚜、铁倍蚜属的4个种(亚种)分别以很高的置信度聚为一支,铁倍花蚜属的铁倍花蚜、小铁枣蚜属的红小铁枣蚜与倍蚜属的角倍蚜和倍蛋蚜聚为一支,但是其置信度较低.同时,采用生物信息学方法从GenBank下载五倍子蚜第一寄主盐肤木属Rhus植物cpDNATrnL-F区序列(980 bp),建立寄主植物种之间的MP和ML系统发育关系,与角倍蚜系统发育关系进行比较分析,结果显示二者之间明显的协同进化关系,但是五倍子蚜的聚类并没有完全和寄主植物之间形成一对一的平行对应关系.     

中国柏木属(Cupressus L.)植物的petG-trnP序列分析及其系统学意义

对中国柏科（Cupressaceae）柏木属（Cupressus L．）5个种及扁柏属4个种的cpDNA非编码区petG—trnP序列进行了分析,其片段的长度范围在417bp～479bp之间．以侧柏（Platycladus orientalis）作为功能性外类群,运用PAUP＊和MEGA软件分别对其进行聚类分析,应用最大似然法得到一棵最优系统树,其-Ln=2232．47．结果表明：用petG-trnP序列来分析柏木属和扁柏属的系统发育关系是可行的;中国柏木属植物是一个单系类群;Ctmmaecyparis nootkatensis与扁柏属其它植物关系较远．因此,柏木（Cupressus funebris）和Ch．nootkatensis均应并入柏木属．     

Phylogenetic positions of some genera and species of the family Buccinidae (Gastropoda: Mollusca) from China based on ribosomal RNA and COI sequences

A phylogenetic analysis of members of the family Buccinidae was conducted using 18S rRNA gene, 28S rRNA gene and the mitochondrial cytochrome oxidase Ⅰ gene. We studied 18 species of Buccinidae that belong to eight different genera and inhabit the China coastal seas. We analyzed the patterns of divergence between an outgroup and basal ingroup taxa, the monophyly of the genus Neptunea, and the position of one unnamed species within the Buccinidae. A phylogenetic tree (neighbor-joining (NJ) method) was reconstructed based on the sequences of 18S rRNA, 28S rRNA and COI, with Rapana venosa as outgroup. The NJ tree indicated that the 18 species could be divided into five groups. The genus Buccinum was monophyletic, whereas Neptunea was shown to be paraphyletic since it included Siphonalia subdilatata and Neptunea sp., a new species. This novel species otherwise clustered consistently with Neptunea cumingi in three other phylogenetic trees, showing a low genetic distance and divergence percentage of sequences belonging to the genus Neptunea. A smaller genetic distance and a smaller difference of 18S rRNA, 28S rRNA and COI sequences between Neptunea cumingii and Neptunea arthritica cumingii confirmed them to be the same species.     

基于PsbA-trnH,RbcL和trnL-F序列的野生葡萄系统发育

采用PCR技术从湘西地区的3种野生葡萄中分离PsbA-trnH、RbcL和trnL-F基因片段,克隆至pMD18-T载体中测序,结合GenBank/EMBL/DDBJ中公布的其他葡萄科植物相应cpDNA基因信息,使用Mega4构建系统发育进化树,分析其系统发育关系.结果显示,PsbA-trnH,RbcL和trnL-F这3种cpDNA基因序列均能用于葡萄科属间的分子鉴定,但结果与传统形态学分类存在一些差异,RbcL能用于葡萄属内种间的分子鉴定.     

中国乳嘴衣属地衣分类学研究

乳嘴衣属为壳状地衣,其所属的乳嘴衣科为热带地区树皮上仅次于文字衣科的第二大丰富类群,目前中国对该属尚无系统性研究。通过处理广东采集的乳嘴衣属标本,扩增其ITS和LSU序列,构建系统发育树,揭示了该属的亲缘关系,并给出了该属中国报道种的相关描述和讨论,以及中国报道种的检索表。     

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.     

基于RAPD标记的15种嵩草属(Kobresia Willd.)植物遗传关系

嵩草属(Kobresia Willd.)植物由于其形态结构的多样性及种内变异式样的复杂性,至今仍被认为是分类学上的"困难属".因此,对于该属系统学其他方面的证据,尤其分子方面的研究则显得极为重要和迫切.本文选用21个RAPD随机引物调查了15种嵩草属植物的遗传关系.结果表明:15种嵩草属植物聚为3类,认为嵩草组和异穗嵩草组的建立是比较自然的,而单穗嵩草组的划分并不自然.此外,分子结论并不支持前人发表的K.maquensis为独立新种,建议降为K.setschwanensis的异名.     

鸡油菌,喇叭菌和钉菇的25SrDNA部分序列及其系统学关系

从分子系统学角度出发,对鸡油菌属（Ｃａｎｔｈａｒｅｌｕｓ）、喇叭菌属（Ｃｒａｔｅｒｅｌｕｓ）和钉菇属（Ｇｏｍｐｈｕｓ）的模式种的２５ＳｒＤＮＡ部分序列进行了测定,并与另外１０目的１３个担子菌和１个接合菌的２５ＳｒＤＮＡ序列进行比较分析,以接合菌总状毛霉（Ｍｕｃｏｒｒａｃｅｍｏｓｕｓ）为外类群构建出这些担子菌的部分２５ＳｒＤＮＡ序列的系统树图,探讨它们３属之间及它们与其他担子菌的关系及其系统学上的意义．认为鸡油菌与同科的喇叭菌关系最为亲近,而钉菇属看来也应该归入鸡油菌目Ｃａｎｔｈａｒｅｌａｌｅｓ中,这与传统方法的分析结果较为一致;虽然鸡油菌类真菌又与各肉质的伞菌类群的关系相对较密切,但它应该有其相对独立的进化路线,将这一类真菌上升至目一级的分类阶元是比较恰当的;但钉菇目Ｇｏｍｐｈａｌｅｓ上升到目的水平是否恰当则有待更多的数据去证明．     

Testing relative evolutionary rates and estimating divergence times among six genera of Rhizophoraceae using cpDNA and nrDNA sequences

The molecular phylogenetic trees of 10 species representing 6 genera of the family Rhizophoraceae have been constructed using the sequences of chloroplast genesmatK andrbcL as well as the ITS regions of nuclear ribosomal DNA. Relative-rate tests between lineages in these phylogenetic trees have been performed. On the basis of the results of the relative-rate tests and related molecular evolutionary rate data, the divergence times between the lineages are estimated as follows: (i) the first divergence time in these genera is 132.25 million years ago (mya); (ii) the average divergence time between two tribes, i.e. inland Legnotideae (exceptCarallia brachiata) and mangrove Rhizophoreae, is 64.13 mya; and (iii) the average divergence time between two inland species,C. garciniaefolia andC. pectinifolia, is 19.92 mya.     

Mitochondrial cytochrome b sequence variations and phylogeny of the East Asian bagrid catfishes

The mitochondrial DNA cytochrome b gene was sequenced from 8 bagrid catfishes in China. Aligned with cytochrome b sequences from 9 bagrid catfishes in Japan, Korea and Russia retrieved from GenBank, and selected Silurus meridionalis, Liobagrus anguillicauda, Liobagrus reini and Phenacogrammus interruptus as outgroups, we constructed a matrix of 21 DNA sequences. The Kimura's two-parameter distances were calculated and molecular phylogenetic trees were constructed by using the maximum parsimony (MP) and neighbor-joining (NJ) methods. The results show that (i) there exist 3-bp deletions of mitochondrial cytochrome b gene compared with cypriniforms and characiforms; (ii) the molecular phylogenetic tree suggests that bagrid catfishes form a monophyletic group, and the genus Mystus is the earliest divergent in the East Asian bagrid catfishes, as well as the genus Pseudobagrus is a monophyletic group but the genus Pelteobagrus and Leiocassis are complicated; and (iii) the evolution rate of the East Asian bagrids mitochondrial cytochrome b gene is about 0.18%～0.30% sequence divergence per million years.     

响叶杨（杨属）叶绿体基因组测序与比较分析

本研究中,我们对响叶杨的叶绿体基因组进行测序和组装,并将其与杨属其他11个叶绿体基因组进行比较分析.结果表明,响叶杨叶绿体基因组全长158,591bp,其中两个反向重复序列区(IR)长度均为27,667bp,长单拷贝序列区(LSC)和短单拷贝序列区(SSC)长度分别为84,634和18,623bp.通过对杨属12个物种的叶绿体基因组进行比较,只发现了6个相对较大的插入缺失,因此整体而言,杨属的叶绿体基因组结构是高度保守的.系统发育分析结果显示,杨属中12个物种组成了3个具有高支持率的进化支,响叶杨与其他白杨组物种聚为一支,并且与银白杨的亲缘关系最近.本研究基于叶绿体基因组数据揭示了杨属的进化历史,将有助于进一步开展杨属植物基于叶绿体DNA序列数据的群体遗传学及其他分子生态学研究.     

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.     

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 specific 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 amplifying 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.     

医蛭形亚目常见物种系统发育基因组学研究

本研究基于GenBank中10个医蛭形亚目物种及2个近缘外类群的基因组或转录组数据,提取COI、18 S和蛋白编码核基因的直系同源序列（Orthologs）,开展系统发育分析。比对后的COI、18 S和Orthologs序列长度分别为1533、1832、220095 bp。最大似然法和贝叶斯推断法分析显示,基于Orthologs序列构建的系统发育树在几乎所有节点上的支持率都达到100%,明显优于COI和18 S序列得到的结果。研究表明,医蛭科的欧洲医蛭、东方医蛭和侧纹医蛭组成一个单系群,医蛭科的日本医蛭与黄蛭科的宽体金线蛭和尖细金线蛭组成另一单系群,而黄蛭科的马蛭与上述6个物种形成姐妹群。医蛭科的两种牛蛭（菲牛蛭和棒纹牛蛭）组成一个单系群,此单系群与医蛭科的其他物种的遗传关系比马蛭更疏远;山蛭科的洞穴山蛭在10个医蛭形亚目种类中处于最基部位置。本研究阐明了医蛭形亚目常见物种的系统发育关系,为这些物种的资源开发利用和保护提供科学依据。