蝗总科部分种类16S rDNA的分子系统发育关系

将自测的我国直翅目蝗总科8科8个种和从互联网GenBank中检索到相关物种的线粒体基因组：16S rDNA序列片段进行同源性比较,计算核苷酸使用频率,并构建分子系统树。在获得的480bp的序列中。A T约占70．7％,G C为29．3％,颠换取代(transversion)的速率大于或接近转换取代(transition)的速率,其中188个核苷酸位点存在变异。研究结果表明：在直翅目蝗总科有差异的188bp中,属内种间的碱基序列差异仅为1．5％,科内属间为3．5％～3．6％,科间差异为4．8％～15．8％,亚目间差异达到15．2％～25．6％。分子系统树表明：科内的属和属内的种均优先聚在一起;蝗总科8科的起源关系为：锥头蝗科→瘤锥蝗科→癞蝗科→斑翅蝗科→剑角蝗科→网翅蝗科和槌角蝗科→斑腿蝗科;锥头蝗科与瘤锥蝗科关系较近,是蝗总科内最原始的类群;槌角蝗科和网翅蝗科互为姐妹群,与最进化的斑腿蝗科关系较近;蚤蝼科为独立的一支,最先分出,似为一个亚目,与现用的分类系统有明显差别;哈螽科(螽嘶总科)和蟋蟀科聚在一起为剑瓣亚目(Ensifera),蚱科和蝗总科的8科组成短瓣亚目(Caehfera),同现用的分类系统。     

蝗科高级阶元的分子系统发育(英文）

迄今,蝗科内各分类阶元之间的系统发生关系大部分是未知的。本文用来自中国24种蝗科昆虫的12SrDNA和16SrDNA2个基因的联合序列(共795bp)数据,以锥头蝗科的锥头蝗(Pyrgomorpha conica)为外群,重建了分子系统树。研究结果表明,在12SrDNA与16SrDNA组成的联合数据中,转换的替代速率明显比颠换的替代速率高得多,核酸的替代已经发生了饱和。分子系统树表明:斑翅蝗亚科是一单系群,该亚科是一个合法的亚科,但斑腿蝗亚科和蝗亚科都不是单系群;斑翅蝗亚科在蝗科内是一个相对原始的类群,而稻蝗亚科比斑翅蝗亚科相对进化,比蝗科的其他亚科的种类相对原始。     

基于COⅡ基因序列的斑腿蝗科部分亚科的分子系统学研究

采用PCR产物直接测序法测定了斑腿蝗科10个亚科16属22种的COⅡ基因585 bp的片段, 对序列的碱基组成进行了分析,并评估了数据集的系统发育信号;最后,以癞蝗科的肃南 短鼻蝗作为外群,采用NJ法、MP法、ML法以及贝叶斯推论法构建了系统树,以解决这些物种所代表的亚科之间的系统发育关系。结果表明：22种斑腿蝗科昆虫的COⅡ基因序列碱基组成表现强烈的A+T含量偏向性。对COⅡ基因585 bp序列片段构成的全数据组和根据密码子不同位点划分的密码子第一、第二和第三位点数据组的系统发育信号分析显示,所有数据组都具有一定的系统发育信息。在4种方法得到的合一树中发现： （1）星翅蝗亚科、刺胸蝗亚科、黑背蝗亚科、斑腿蝗亚科的亲缘关系较近;（2）卵翅蝗亚科与稻蝗亚科亲缘关系较近,建议卵翅蝗亚科似乎应归入稻蝗亚科中,板胸蝗亚科与这两个亚科的关系较近;（3）黑蝗亚科和秃蝗亚科似乎应合并为一个亚科;（4）切翅蝗亚科的4个属未聚在一起,表明这些属的区别较大,不是一个单系群;（5）黑蝗亚科和秃蝗亚科关系较近,且与本研究中其他几个亚科的亲缘关系相对较远。研究结果表明COⅡ基因在解决斑腿蝗科的亚科以下属种间的系统发育关系时是一个有效的分子标记。     

蝗总科部分种类Cyt b基因序列及系统进化研究

蝗总科8个科10个种的线粒体DNA细胞色素b(Cyt b）基因部分序列被测定分析,比较同源性,计算核苷酸使用频率并构建NJ分子系统树。在获得的432bp的序列中,A T约占70.4%,其中177个核苷酸位点存在变异（约占41.0%）,DNA一级序列数据显示,该8科蝗虫间DNA序列变异丰富。用日本蚱作外群构建的NJ树表明：瘤锥蝗科较为原始,而癞蝗科与槌角蝗科关系较近,为比较进化的类群,蝗总科8科的起源关系为：瘤锥蝗科→班翅蝗科和锥头蝗科→斑腿蝗科→剑角蝗科→网翅蝗科→槌角蝗科和癞蝗科,上述研究结果与传统形态分类学结论不完全一致,并就此进行了讨论。     

基于18S rRNA基因序列的直翅目主要类群系统发育关系研究

基于78种直翅目昆虫的18S rRNA基因全序列构建了直翅目各主要类群间的系统发育关系。本研究的结果支持直翅目的单系性,但不支持蝗亚目和螽亚目各自的单系性;直翅目下除蜢总科和蝗总科外各总科的划分多数与Otte系统相一致;蜢总科的单系性得不到支持;蝗总科的剑角蝗科、斑腿蝗科、斑翅蝗科、网翅蝗科和槌角蝗科5科均不是单系群,各物种间的遗传距离差异不大,应合并为一科,即蝗科;本研究支持将Otte系统中蚱总科和螽蟖总科下各亚科级阶元提升为科级阶元;18S rRNA基因全序列可以作为划分科级阶元的工具,当位于同一分支上互成姐妹群的类群间的遗传距离超过1%时,这几个类群属于不同的科;但由于其在进化上的保守性,18S rRNA基因只能用于纲目等高级阶元间关系的研究,而由其获得的总科以下阶元间的关系并不可靠。     

基于线粒体Cyt b,16S rDNA和核28S rDNA的中国斑腿蝗科(直翅目,蝗总科)九亚科间的系统发育关系研究

基于线粒体Cytb、16S rDNA和核28S rDNA的部分序列的联合数据集,重建了中国斑腿蝗科9亚科26种蝗虫和癞蝗科笨蝗(外群)间的系统发育关系。结果显示:1)除黑蝗亚科、秃蝗亚科和切翅蝗亚科外,其余6亚科的单系性都得到支持;2)黑蝗亚科似乎应当与秃蝗亚科合并为一个亚科,且与其他几个亚科亲缘关系相对较远;3)凸额蝗属应该由切翅蝗亚科中独立出来;4)卵翅蝗亚科和稻蝗亚科是姐妹群;5)星翅蝗亚科、黑背蝗亚科、斑腿蝗亚科、刺胸蝗亚科及切翅蝗亚科中的凸额蝗属间的关系较近。     

基于线粒体ND2基因的中国斑翅蝗科部分种类分子系统学研究（直翅目：蝗总科）

本文的目的是通过对斑翅蝗科部分种类的线粒体ND2基因进行分析,重建斑翅蝗科昆虫的系统发育关系,并探讨分子系统发育关系和传统分类结果的异同。扩增并测定了我国斑翅蝗科10属16种蝗虫的线粒体ND2全基因1 023 bp的序列,对序列的碱基组成、转换颠换、系统发育信号等进行了分析。并基于ND2全基因序列数据,分别采用邻接法（NJ）、最简约法（MP）、最大似然法（ML）和贝叶斯法重建了10属16种蝗虫的系统发育关系。结果表明：斑翅蝗科蝗虫ND2全基因A+T含量平均为74.6%;痂蝗亚科和异痂蝗亚科没能得到区分,建议合并为一个亚科;而斑翅蝗亚科和飞蝗亚科的分类地位还存在争议。     

蝗总科精子超微形态比较及其在系统分类上的意义（直翅目）

通过对８科１９属２３种蝗虫精子超微形态结构的观察和比较,发现其精子有４种类型。这４种类型精子间的主要差异反映在中心粒侧体及线粒体衍生体的超微形态结构方面。经过比较和分析,确认８科中癞蝗科种类（除友谊华癞蝗外）的精子最原始,斑腿蝗科、斑翅蝗科、网翅蝗科、槌角蝗科、剑角蝗科（除中华蚱蜢外）种类的精子最进化。根据精子的演化类型建立了蝗总科精子系统,在这个系统里蝗总科８科昆虫在演化趋势上可分为３大类群     

基于Cyt b基因的网翅蝗科系统发生重建

研究测定了蝗总科25种蝗虫的线粒体Cyt b部分序列,并从GenBank中下载了19种蝗亚目昆虫的Cyt b基因相应序列片段。本文目的是要建立网翅蝗科的系统发育关系并说明网翅蝗科在蝗总科中的分类地位。以瘤锥蝗科的云南蝗Yunnanites coriacea和长额橄蝗Tagasta marginella作为外群,用MP法和贝叶斯法重建系统发生树。比对后的序列长度是384 bp,包括167个简约信息位点。A T平均含量为70.7%,C G 平均含量29.3%。分子系统树表明:网翅蝗科并不是一个单系群。网翅蝗亚科和竹蝗亚科并非单系群。现存的雏蝗属并非单系群,应该是多系群。分子系统学研究结果和传统的基于形态特征的网翅蝗科分类体系有很大的不同。     

基于Cytb和COⅡ基因序列的网翅蝗科(直翅目:蝗总科)部分种类的分子系统学研究

采用PCR产物直接测序法测定了2亚科31种网翅蝗科昆虫及1种癞蝗(外群)的线粒体基因Cyt b和COⅡ全序列,使用MEGA V4.1进行序列组成分析,对线粒体基因Cyt b、COⅡ数据集按照蛋白质基因密码子第一、二、三位点划分子集,并组成联合数据集COⅡ&Cyt b,对各数据集其子集进行了数据探索研究.在PAUP中应用NJ法、MP法、ML法以及在MrBayes V3.1中应用贝叶斯系统发育推论(BI)法进行系统发育关系重建,结果表明所研究的网翅蝗科部分种类的系统发育关系基本与中国的传统形态学分类体系没有差别,竹蝗亚科为较早分化出来的类群,其次为网翅蝗亚科;竹蝗新种和黄脊雷篦蝗聚为一支和其它竹蝗属类群形成姊妹群,因此建议将雷篾蝗属并入竹蝗属;隆额网翅蝗和宽翅曲背蝗聚为另一支,故建议将曲背蝗属并入网翅蝗属.雏蝗属的四个亚属中黑翅亚属、曲隆亚属和直隆亚属这三个亚属的单系性均得到稳健的支持,而短翅亚属的单系性在各种分析中皆得不到支持,它很可能是多系起源.     

基于18S rDNA的蝗总科分子系统发育关系研究及分类系统探讨

将自测的我国直翅目蝗总科7科7种和从GenBank中下载的17种直翅目昆虫的18S rDNA序列片段进行了同源性比较,用似然比检验的方法对序列比对结果进行了碱基替代模型的选择,以蚱总科的Paratettix cucullatus和蜢总科的Stiphra robusta作外群,用NJ、MP、ML和贝叶斯法构建了分子系统树。在获得的1 849 bp的序列中,有205个变异位点,74个简约信息位点; A、T、C和G的碱基平均含量分别为23.9%、24.3%、23.8%和28.0%,碱基组成基本上无偏异。分子系统树表明：所研究的内群聚为4支,锥头蝗科、瘤锥蝗科、斑腿蝗科、网翅蝗科、槌角蝗科和剑角蝗科都不是单系。建议将蝗总科分为4科,即锥头蝗科、大腹蝗科、癞蝗科和蝗科。     

New taxa refresh the phylogeny and classification of pleurostomatid ciliates (Ciliophora,Litostomatea)

A high diversity of pleurostomatid ciliates has been discovered in the last decade, and their systematics needs to be improved in the light of new findings concerning their morphology and molecular phylogeny. In this work, a new genus, Protolitonotus gen. n., and two new species, Protolitonotus magnus sp. n. and Protolitonotus longus sp. n., were studied. Furthermore, 19 novel nucleotide sequences of SSU rDNA, LSU rDNA and ITS1‐5.8S‐ITS2 were collected to determine the phylogenetic relationships and systematic positions of the pleurostomatid ciliates in this study. Based on both molecular and morphological data, the results demonstrated that: (i) as disclosed by the sequence analysis of SSU rDNA, LSU rDNA and ITS1‐5.8S‐ITS2, Protolitonotus gen. n. is sister to all other pleurostomatids and thus represents an independent lineage and a separate family, Protolitonotidae fam. n., which is defined by the presence of a semi‐suture formed by the right somatic kineties near the dorsal margin of the body; (ii) the families Litonotidae and Kentrophyllidae are both monophyletic based on both SSU rDNA and LSU rDNA sequences, whereas Amphileptidae are non‐monophyletic in trees inferred from SSU rDNA sequences; and (iii) the genera Loxophyllum and Kentrophyllum are both monophyletic, whereas Litonotus is non‐monophyletic based on SSU rDNA analyses. ITS1‐5.8S‐ITS2 sequence data were used for the phylogenetic analyses of pleurostomatids for the first time; however, species relationships were less well resolved than in the SSU rDNA and LSU rDNA trees. In addition, a major revision to the classification of the order Pleurostomatida is suggested and a key to its families and genera is provided.     

Phylogenetic analysis of ribosomal RNA operons from uncultivated coastal marine bacterioplankton

Analyses of small subunit ribosomal RNA genes (SSU rDNAs) have significantly influenced our understanding of the composition of aquatic microbial assemblages. Unfortunately, SSU rDNA sequences often do not have sufficient resolving power to differentiate closely related species. To address this general problem for uncultivated bacterioplankton taxa, we analysed and compared sequences of polymerase chain reaction (PCR)-generated and bacterial artificial chromosome (BAC)-derived clones that contained most of the SSU rDNAs, the internal transcribed spacer (ITS) and the large subunit ribosomal RNA gene (LSU rDNA). The phylogenetic representation in the rRNA operon PCR library was similar to that reported previously in coastal bacterioplankton SSU rDNA libraries. We observed good concordance between the phylogenetic relationships among coastal bacterioplankton inferred from SSU or LSU rDNA sequences. ITS sequences confirmed the close intragroup relationships among members of the SAR11, SAR116 and SAR86 clades that were predicted by SSU and LSU rDNA sequence analyses. We also found strong support for homologous recombination between the ITS regions of operons from the SAR11 clade.     

Amplification of mitochondrial small subunit ribosomal DNA of polypores and its potential for phylogenetic analysis

There has been a systematic need to seek adequate phylogenetic markers that can be applied in phylogenetic analyses of fungal taxa at various levels. The mitochondrial small subunit ribosomal DNA (mt SSU rDNA) is generally considered to be one of the molecules that are appropriate for phylogenetic analyses at a family level. In order to obtain universal primers for polypores of Hymenomycetes, mt SSU rRNA genes were cloned from Bjerkandera adusta, Ganoderma lucidum, Phlebiopsis gigantea, and Phellinus laevigatus and their sequences were determined. Based on the conserved sequences of cloned genes from polypores and Agrocybe aegerita, PCR primers were designed for amplification and sequencing of mt SSU rDNAs. New primers allowed effective amplification and sequencing of almost full-sized genes from representative species of polypores and related species. Phylogenetic relationships were resolved quite efficiently by mt SSU rDNA sequences, and they proved to be more useful in phylogenetic reconstruction of Ganoderma than nuclear internal transcribed spacer (ITS) rDNA sequences.     

Phylogenetic analysis of Glomeromycota by partial LSU rDNA sequences

We analyzed the large subunit ribosomal RNA (rRNA) gene [LSU ribosomal DNA (rDNA)] as a phylogenetic marker for arbuscular mycorrhizal (AM) fungal taxonomy. Partial LSU rDNA sequences were obtained from ten AM fungal isolates, comprising seven species, with two new primers designed for Glomeromycota LSU rDNA. The sequences, together with 58 sequences available from the databases, represented 31 AM fungal species. Neighbor joining and parsimony analyses were performed with the aim of evaluating the potential of the LSU rDNA for phylogenetic resolution. The resulting trees indicated that Archaeosporaceae are a basal group in Glomeromycota, Acaulosporaceae and Gigasporaceae belong to the same clade, while Glomeraceae are polyphyletic. The results support data obtained with the small subunit (SSU) rRNA gene, demonstrating that the LSU rRNA gene is a useful molecular marker for clarifying taxonomic and phylogenetic relationships in Glomeromycota.     

Phylogenetic validation of the genera Angomonas and Strigomonas of trypanosomatids harboring bacterial endosymbionts with the description of new species of trypanosomatids and of proteobacterial symbionts

We comparatively examined the nutritional, molecular and optical and electron microscopical characteristics of reference species and new isolates of trypanosomatids harboring bacterial endosymbionts. Sequencing of the V7V8 region of the small subunit of the ribosomal RNA (SSU rRNA) gene distinguished six major genotypes among the 13 isolates examined. The entire sequences of the SSU rRNA and glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) genes were obtained for phylogenetic analyses. In the resulting phylogenetic trees, the symbiont-harboring species clustered as a major clade comprising two subclades that corresponded to the proposed genera Angomonas and Strigomonas. The genus Angomonas comprised 10 flagellates including former Crithidia deanei and C. desouzai plus a new species. The genus Strigomonas included former Crithidia oncopelti and Blastocrithidia culicis plus a new species. Sequences from the internal transcribed spacer of ribosomal DNA (ITS rDNA) and size polymorphism of kinetoplast DNA (kDNA) minicircles revealed considerable genetic heterogeneity within the genera Angomonas and Strigomonas. Phylogenetic analyses based on 16S rDNA and ITS rDNA sequences demonstrated that all of the endosymbionts belonged to the Betaproteobacteria and revealed three new species. The congruence of the phylogenetic trees of trypanosomatids and their symbionts support a co-divergent host-symbiont evolutionary history.     

Phylogenetic relationships of Palaeacanthocephala (Acanthocephala) inferred from SSU and LSU rDNA gene sequences

The Palaeacanthocephala is traditionally represented by 2 orders, Echinorhynchida and Polymorphida, with 10 and 3 families, respectively. To test the monophyly of the class, these 2 orders, and certain families, phylogenies were inferred using nuclear small-subunit (SSU) and large-subunit (LSU) ribosomal DNA sequences obtained for 29 species representing 10 families, 2 other classes of acanthocephalans, and 3 rotifer outgroups. Phylogenetic relationships were inferred by analyzing combined SSU and LSU sequences using maximum parsimony (MP) and maximum likelihood (ML) methods. Parsimony and ML trees inferred from combined analysis of these rDNA data strongly supported monophyly of Palaeacanthocephala and provided good resolution among species. Neither Polymorphida nor Echinorhynchida was monophyletic. Gorgorhynchoides bullocki (Echinorhynchida) was nested within the 6 species representing Polymorphida, and this clade was nested within species representing Echinorhynchida. Three of 4 palaeacanthocephalan families that could be evaluated were not monophyletic, and this finding was strongly supported. These results indicate that the family level classification of palaeacanthocephalans, which is mainly based on combinations of shared characters (not shared derived characters), needs to be reevaluated with respect to comprehensively sampled phylogenetic hypotheses.     

Nuclear-encoded rDNA group I introns: origin and phylogenetic relationships of insertion site lineages in the green algae

Group I introns are widespread in eukaryotic organelles and nuclear- encoded ribosomal DNAs (rDNAs). The green algae are particularly rich in rDNA group I introns. To better understand the origins and phylogenetic relationships of green algal nuclear-encoded small subunit rDNA group I introns, a secondary structure-based alignment was constructed with available intron sequences and 11 new subgroup ICI and three new subgroup IB3 intron sequences determined from members of the Trebouxiophyceae (common phycobiont components of lichen) and the Ulvophyceae. Phylogenetic analyses using a weighted maximum-parsimony method showed that most group I introns form distinct lineages defined by insertion sites within the SSU rDNA. The comparison of topologies defining the phylogenetic relationships of 12 members of the 1512 group I intron insertion site lineage (position relative to the E. coli SSU rDNA coding region) with that of the host cells (i.e., SSU rDNAs) that contain these introns provided insights into the possible origin, stability, loss, and lateral transfer of ICI group I introns. The phylogenetic data were consistent with a viral origin of the 1512 group I intron in the green algae. This intron appears to have originated, minimally, within the SSU rDNA of the common ancestor of the trebouxiophytes and has subsequently been vertically inherited within this algal lineage with loss of the intron in some taxa. The phylogenetic analyses also suggested that the 1512 intron was laterally transferred among later-diverging trebouxiophytes; these algal taxa may have coexisted in a developing lichen thallus, thus facilitating cell- to-cell contact and the lateral transfer. Comparison of available group I intron sequences from the nuclear-encoded SSU rDNA of phycobiont and mycobiont components of lichens demonstrated that these sequences have independent origins and are not the result of lateral transfer from one component to the other.      

Phylogenetic analysis of the Metastrongyloidea (Nematoda: Strongylida) inferred from ribosomal RNA gene sequences

Phylogenetic relationships among nematodes of the strongylid superfamily Metastrongyloidea were analyzed using partial sequences from the large-subunit ribosomal RNA (LSU rRNA) and small-subunit ribosomal RNA (SSU rRNA) genes. Regions of nuclear ribosomal DNA (rDNA) were amplified by polymerase chain reaction, directly sequenced, aligned, and phylogenies inferred using maximum parsimony. Phylogenetic hypotheses inferred from the SSU rRNA gene supported the monophyly of representative taxa from each of the 7 currently accepted metastrongyloid families. Metastrongyloid taxa formed the sister group to representative trichostrongyloid sequences based on SSU data. Sequences from either the SSU or LSU RNA regions alone provided poor resolution for relationships within the Metastrongyloidea. However, a combined analysis using sequences from all rDNA regions yielded 3 equally parsimonious trees that represented the abursate Filaroididae as polyphyletic, Parafilaroides decorus as the sister species to the monophyletic Pseudaliidae, and a sister group relationship between Oslerus osleri and Metastrongylus salmi. Relationships among 3 members of the Crenosomatidae, and 1 representative of the Skrjabingylidae (Skrjabingylus chitwoodorum) were not resolved by these combined data. However, members of both these groups were consistently resolved as the sister group to the other metastrongyloid families. These relationships are inconsistent with traditional classifications of the Metastrongyloidea and existing hypotheses for their evolution.