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。     

中国脊筒天牛属分类研究(鞘翅目:天牛科)

系统整理了中国脊筒天牛属失一部种类,共31种,给出了所有种类的分类检索表,记述了新种暗背脊筒天牛Nupserha fuscodorsalis Wang et Chiang,sp nov。及2中国新纪录种三斑脊筒天牛Nupserha multimaculata Pic和黑肩脊筒天牛Nupserha nigrohumeralis Pic,新种正模保存于西南农业大学昆虫标本馆,副模共5号存于南开大学生物系标本室。     

星天牛属的数值分类研究初探(鞘翅目:天牛科)

本文选取了92个综合特征对星天牛属 Anoplophora 的九个种分别进行了系统聚类法和主成分分析研究,结果是一致的。灰星天牛 A.versteegi(Ritsema)是一个比较特殊的种,在体型和一系列特征上都与其它星天牛类存在较大的差异,因此把它从本属中分出,归入安天牛属Annamanum是比较合理的。许多学者将 Anoplophora分开成两个亚属,甚或两个不同的属,数值分类结果表明,除A.versteegi外的所有种中,不存在明显的类元划分界线,且也不存在秩级划分的重要特征,而是呈均一的自然类群。 用三个相似性系数借助于三个协表相相关系数作为最优性分类的评判标准对系统聚类法的九种聚合策略进行了比较研究,结果表明UPGMA是最佳的。     

沟胫天牛族三新种记述(鞘翅目:天牛科:沟胫天牛亚科)

记述了沟胫天牛亚科沟胫天牛族Ａｇｎｉｎｉ３新种,即双斑齿胫天牛Ｐａｒａｌｅｐｒｏｄｅｒａ ｂｉｍａｃｕｌａｔａ ｓｐ．ｎｏｖ．,云南棱柄天牛Ｎａｎｏｈａｍｍｕｓ ｙｕｎｎａｎａ ｓｐ．ｎｏｖ．和白斑肖墨天牛Ｘｅｎｏｈａｍｍｕｓ ａｌｂｏｍａｃｕｌａｔａ ｓｐ．ｎｏｖ．。新种模式标本保存于西南农业大学昆虫标本馆。     

鞘翅目昆虫核酸分子系统学研究现状

从研究对象、研究种类、研究内容等方面对鞘翅目Coleoptera核酸分子系统学研究的近况进行了总结和分析,研究中应用的技术主要有DNA序列分析、RELP技术、RAPD技术、AFLP技术、分子杂交技术和SSCD技术。并认为这些技术的应用对补充和完善传统分类方法,深入探讨各类群的分类地位和系统发育关系具有重要作用。     

小鲵科线粒体16S rRNA基因序列分析及其系统发育

To study the phylogeny of Hynobiidae, we amplified DNA fragments of 470 bp 16S ribosomal RNA (16S rRNA) gene on mitochondrial DNA from Ranodon sibiricus and Ranodon tsinpaensis. PCR products were cloned into PMD18 T vector after purification. These sequences were determined and deposited in the GenBank (accession numbers: AY373459 for Ranodon sibiricus, AY372534 for Ranodon tsinpaensis). By comparing the nucleotide differences of 16S ribosomal RNA sequences among Liua shihi, Pseudohynobius flavomaculatus and Batrachuperus genus from GenBank database, we analyzed the divergences and base substitution among these sequences with the MEGA software. The molecular results support that B. tibetanus, B. pinchonii and B. karlschmidti are classified into three valid species. Liua shihi has closer phylogenetic relationships to Ranodon tsinpaensis than to other species. More our results reveal that Pseudohynobius flavomaculatus is not a synonym of Ranodon tsinpaensis. [Acta Zoologica Sinica 50 (3) : 464 - 469,2004].     

基于28S rDNA 的叩甲科分子系统发育关系研究

【目的】通过对叩甲科（Elateridae）昆虫核糖体28S rDNA基因片段序列进行比较,从分子水平研究叩甲科昆虫的系统发育关系,并和传统分类结果相比较,为我国叩甲科分类系统的论证和进一步修订奠定基础。【方法】将自测的我国9种（含两个地理种群）共10个叩甲科昆虫样品的28S rDNA基因片段序列与GenBank报道的32种叩甲科昆虫进行同一性比较,用DNAStar Lasergene v 7.1.0和MEGA4.0（NJ法、MP法和ME法）构建分子系统发育树。【结果】在获得的890 bp的序列中,保守位点477个,占全部位点的56.1％;简约位点291个,占全部位点的34.2％;G+C的平均含量为63.9％,明显高于A+T的平均含量,碱基组成偏向G和C;转换（transition）稍高于颠换（transversion）。遗传距离分析表明叩甲科昆虫各亚科内各种间遗传距离在0.000～0.130之间变动,明显小于各亚科之间的遗传距离。不同的系统发育树都支持叩甲科为一单系群,并将10个亚科聚为4个聚类簇:聚类簇Ⅰ为梳爪叩甲亚科（Melanotinae）＋叩甲亚科（Elaterinae）,聚类簇Ⅱ为槽缝叩甲亚科（Agrypninae）＋萤叩甲亚科（Pyrophorinae）＋单叶叩甲亚科（Conoderinae）,聚类簇Ⅲ为小叩甲亚科（Negastriinae）＋心盾叩甲亚科（Cardiophorinae）,聚类簇Ⅳ为齿胸叩甲亚科（Denticollinae）＋尖鞘叩甲亚科（Oxynopterinae）和异角叩甲亚科（Pityobiinae）。它们来源于2个支系,支系1包含聚类簇Ⅰ,支系2包含聚类簇Ⅱ、聚类簇Ⅲ和聚类簇Ⅳ,而Senodonia quadricollis总是单独作为一支与其他叩甲分开。【结论】本研究证实了过去基于成虫和幼虫形态为基础的分类系统的基本合理性,一是叩甲科为一单系类群;二是叩甲科可明显地分为4个簇群;三是心盾叩甲亚科（Cardiophorinae）为一单系类群,但其他许多亚科存在并系的情况,特别是Senodonia quadricollis的归属还需进一步论证。28S rDNA 序列分析是一种很好的研究叩甲科从种级到科级各类群间的系统发育关系的方法。     

基于核糖体DNA联合序列的天牛总科高阶元分子系统学研究

【目的】利用核糖体DNA联合序列探讨天牛总科高阶元分子系统发育。【方法】本研究采用分子标记技术,分析测定了63种天牛核糖体28S rDNA D2和D3区以及18S rDNA V4和V7区的DNA序列,并采用邻接法、最大似然法和贝叶斯推论法分别构建了天牛总科2科6亚科63种的分子进化系统。【结果】序列联合比对分析,最终得到1 404 bp的联合数据组,其中可变位点446个（32.0%）,保守位点958（68.0%）,转换/颠换的平均值（R值）为1.73。28S rDNA和18S rDNA以及联合序列的饱和度分析显示碱基突变未达到饱和,说明这些序列适合于分子进化树的构建。利用不同系统发育重建方法得到进化树具有相似拓扑结构,结果支持沟胫天牛亚科、花天牛亚科和天牛亚科为单系群,这与形态学分类结果相似;狭胸天牛独立成为亚科得到了支持。【结论】利用28S rDNA D2和D3区以及18S rDNA V4和V7区联合序列成功构建出了天牛总科高阶元的系统发育树。研究表明联合序列分析是探讨天牛高阶元分类的有效的方法。     

天牛亚科中国四新纪录种(鞘翅目,天牛科)

记述了天牛亚科虎天牛族4新纪录种,刺虎天牛属Demonax Thomson的八点刺虎天牛D.contrarius Holzschuh;艳虎天牛属Rhaphuma Pascoe的独艳虎天牛R.unigena Holzschuh;绿虎天牛属Chlorophorus Chevrolat的胖绿虎天牛C.proannulatus Gressitt & Rondon;瘦棍腿天牛属Stenodryas Bates的四纹瘦棍腿天牛S.nigromaculatus(Gardner).研究标本保存在西南大学昆虫标本馆.     

Molecular Phylogeny of the Family Ophryscolecidae (Ciliophora,Litostomatea) Inferred from 18S rDNA Sequences

The 18S rDNA was used to infer oral ciliature patterns of evolution within the family Ophryoscolecidae, with the addition of five new sequences of ciliates from the genus Ostracodinium. Our data confirmed the monophyly of the subfamilies Entodiniinae and Ophryoscolecinae, but more analysis would be required for the definition of the status of the subfamily Diplodiniinae. The oral infraciliature patterns reflect evolutionary divergence in the family Ophryscolecidae, observing monophyly on Entodinium‐type, Diplodinium‐type, Ostracodinium‐type, Epidinium‐type, and Ophryoscolex‐type. The ancestral infraciliature of Entodinium‐type cannot be proven, however, the position of Entodinium‐type showed closer of Diplodinium‐type than Ophryoscolex‐type, corroborating previous studies using morphological characters. The observed inconsistencies reflect the need to increase the number of 18S rDNA sequences to family Ophryoscolecidae and investigate the evolution of this group using other molecular markers.     

基于16S rDNA和28S rDNA D2基因序列与形态特征联合分析的中国角顶叶蝉亚科系统发育研究（半翅目： 叶蝉科）

首次在国内利用28S rDNA D2区段和16S rDNA基因序列,结合50个形态特征对角顶叶蝉亚科(Deltocephalinae)［半翅目（Hemiptera）: 叶蝉科（Cicadellidae）］19个属进行系统发育分析研究。从无水乙醇浸泡保存的标本中提取基因组DNA并扩增了19个内群和1种外群Typhlocybinae［半翅目(Hemiptera): 叶蝉科(Cicadellidae)］种类的28S rDNA D2基因片段并测序,同时扩增了16S rDNA基因片段并测序11条,采用了GenBank中1个种类的16S rDNA同源序列。采用PAUP*4.0和MrBayes3.0两个分析软件和3种建树方法,利用同源28S D2 rDNA和16S rDNA两个基因序列与形态特征结合进行系统发育分析研究。分析结果表明,二叉叶蝉族Macrostelini是一个单系,并在角顶叶蝉亚科的系统发育中处于基部的位置,是内群中最原始的族;角顶叶蝉族Deltocephalini中除了纹翅叶蝉属Nakaharanus,其余各属构成单系;殃叶蝉族Euscelini内属的归属比较混乱,可能是一个并系群,属间差异有待进一步研究。隆额叶蝉族Paralimnini与顶带叶蝉族Athysanini是姐妹群。带叶蝉属Scaphoideus与纹翅叶蝉属Nakaharanus是姐妹群,二者与木叶蝉属Phlogotettix的关系最近,三者构成一个单系,建议将三者归为带叶蝉族Scaphoideini。研究结果还表明,小眼叶蝉族Xestocephalini和Balcluthini的系统发育位置不明,有待进一步研究。     

基于18S rDNA序列的蝽次目(半翅目：异翅亚目)

利用18SrDNA分子约1 912 bp的序列对蝽次目21个科53个种进行系统发育分析。运用MP法、ML法和NJ法分析后的结果表明:蝽次目的单系性得到很高的支持;扁蝽总科成为毛点类的姐妹群;毛点类基本确定为两大分支:一支包含蝽总科和红蝽总科;另一支主要由长蝽总科、缘蝽总科和南蝽总科组成;长蝽总科和缘蝽总科都是多系;长蝽总科中,跷蝽科和皮蝽科的关系最近,构成姐妹群,位于整个毛点类的基部;与长蝽总科中另外两个科长蝽科和地长蝽科的关系很远。说明利用18SrDNA分子对研究蝽次目的系统发育关系是适合的,能够重建蝽次目;扁蝽总科和蝽总科单系性的结果与形态学的研究以及Li et al (2005)的研究一致;但较Li et al(2005)的研究更进一步把红蝽总科从广义的缘蝽总科中分出来;并建议皮蝽科作为一个独立的总科更合适。     

烟粉虱内共生菌16S rDNA的特性研究

对烟粉虱内共生菌16SrDNA的酶切结果及部分生物体16SrDNA的(G C)%分析结果表明:烟粉虱初生内共生菌的16SrDNA能够被EcoRI酶切成两个片段、而不能够分别被BamHI与SacI酶切;烟粉虱次生内共生菌的16SrDNA没有BamHI内切酶位点、而能够分别被EcoRI或Sac I酶切成大小不同的两个片段。(G C)mol%与菌的分类地位有关,同时还与菌的可培养能力有关。Proteobacteriaγ亚纲的烟粉虱初生内共生菌与α亚纲的Rickettsia、线粒体的16SrDNA相似,富含(A T)mol%,具低的(G C)mol%。而γ亚纲的次生内共生菌及大肠杆菌与β亚纲的mealybugs初生内共生菌的16SrDNA相似,富含(G C)mol%。说明初生内共生菌可能与烟粉虱同时发生,并且形成一种非常紧密的共生关系,次生内共生菌与烟粉虱关系松散一些,其特性近似于自由生活的细菌,更有可能获得纯培养体。16SrDNA的系统进化树表明,烟粉虱次生内共生菌属于Proteobacteriaγ-3亚纲,而初生内共生菌属于Proteobacteriaγ亚纲的另一分支。     

Molecular Phylogeny of Conjugating Green Algae (Zygnemophyceae, Streptophyta) Inferred from SSU rDNA Sequence Comparisons

Abstract Nuclear-encoded SSU rDNA sequences have been obtained from 64 strains of conjugating green algae (Zygnemophyceae, Streptophyta, Viridiplantae). Molecular phylogenetic analyses of 90 SSU rDNA sequences of Viridiplantae (inciuding 78 from the Zygnemophyceae) were performed using complex evolutionary models and maximum likelihood, distance, and maximum parsimony methods. The significance of the results was tested by bootstrap analyses, deletion of long-branch taxa, relative rate tests, and Kishino–Hasegawa tests with user-defined trees. All results support the monophyly of the class Zygnemophyceae and of the order Desmidiales. The second order, Zygnematales, forms a series of early-branching clades in paraphyletic succession, with the two traditional families Mesotaeniaceae and Zygnemataceae not recovered as lineages. Instead, a long-branch Spirogyra/Sirogonium clade and the later-diverging Netrium and Roya clades represent independent clades. Within the order Desmidiales, the families Gonatozygaceae and Closteriaceae are monophyletic, whereas the Peniaceae (represented only by Penium margaritaceum) and the Desmidiaceae represent a single weakly supported lineage. Within the Desmidiaceae short internal branches and varying rates of sequence evolution among taxa reduce the phylogenetic resolution significantly. The SSU rDNA-based phylogeny is largely congruent with a published analysis of the rbcL phylogeny of the Zygnemophyceae (McCourt et al. 2000) and is also in general agreement with classification schemes based on cell wall ultrastructure. The extended taxon sampling at the subgenus level provides solid evidence that many genera in the Zygnemophyceae are not monophyletic and that the genus concept in the group needs to be revised.     

Phylogeny of coral-inhabiting barnacles (Cirripedia; Thoracica; Pyrgomatidae) based on 12S, 16S and 18S rDNA analysis

The traditional phylogeny of the coral-inhabiting barnacles, the Pyrgomatidae, is based on morphological characteristics, mainly of the hard parts. It has been difficult to establish the phylogenetic relationships among Pyrgomatidae because of the apparent convergence of morphological characteristics, and due to the use of non-cladistic systematics, which emphasize ancestor-descendant relationships rather than sister-clade relationships. We used partial sequences of two mithochondrial genes, 12S rDNA and 16S rDNA, and a nuclear gene, 18S rDNA, to infer the molecular phylogeny of the pyrgomatids. Our phylogenetic results allowed us to reject previous classifications of Pyrgomatidae based on morphological characteristics. Our results also suggested the possibility of paraphyly of the Pyrgomatidae. The hydrocoral barnacle Wanella is not found on the same clade as the other pyrgomatids, but rather, with the free-living balanids. The basal position of Megatrema and Ceratoconcha is supported. The archeaobalanid Armatobalanus is grouped with Cantellius at the base of the Indo-Pacific pyrgomatines. Fusion of the shell plate and modification of the opercular valves are homoplasious features that occurred more than three times on different clades. The monophyly of the "Savignium" group, comprising four nominal genera, is also not supported, and the different taxa are placed on different clades.     

Phylogeny of Molting Protostomes (Ecdysozoa) as Inferred from 18S and 28S rRNA Gene Sequences

Phylogenetic relationships within the group of molting protostomes were reconstructed by comparing the sets of 18S and 28S rRNA gene sequences considered either separately or in combination. The reliability of reconstructions was estimated from the bootstrap indices for major phylogenetic tree nodes and from the degree of congruence of phylogenetic trees obtained by different methods. By either criterion, the phylogenetic trees reconstructed on the basis of both 18 and 28S rRNA gene sequences were better than those based on the 18S or 28S sequences alone. The results of reconstruction are consistent with the phylogenetic hypothesis classifying protostomes into two major clades: molting Ecdysozoa (Priapulida + Kinorhyncha, Nematoda + Nematomorpha, Onychophora + Tardigrada, Myriapoda + Chelicerata, and Crustacea + Hexapoda) and nonmolting Lophotrochozoa (Plathelminthes, Nemertini, Annelida, Mollusca, Echiura, and Sipuncula). Nematomorphs (Nematomorpha) do not belong to the clade Cephalorhyncha (Priapulida + Kinorhyncha). It is concluded that combined data on the 18S and 28S rRNA gene sequences provide a more reliable basis for phylogenetic inferences.__________Translated from Molekulyarnaya Biologiya, Vol. 39, No. 4, 2005, pp. 590–601.Original Russian Text Copyright © 2005 by Petrov, Vladychenskaya.     

基于线粒体16S rRNA基因对中国狼蛛科(Lycosidae)系统发育研究

将自测的中国狼蛛科Lycosidae4亚科6属26种和从GenBank中检索到的北美2种豹蛛的mtD-NA-16S rRNA序列进行比较;以漏斗蛛科1种蜘蛛作为外群,对碱基序列的组成和遗传距离进行了分析,采用Bayesian方法和最大简约法(MP)构建分子系统树。研究结果表明:16SrRNA基因的部分序列为340bp到360bp,A T含量平均为75%,存在较强的A T含量偏向性;序列共有157个碱基存在变异,其中79个简约信息位点。狼蛛科各属间的遗传距离介于0.026￣0.200之间。2种建树方法均表明:科内的属及属内的种优先聚在一起;水狼蛛属相对马蛛属是狼蛛科中较为原始的类群,分化较早;獾蛛属作为1个单系群与熊蛛属合为1个并系,属于狼蛛亚科。狼蛛科6属间的分子系统关系为(Pirata(Hippasa(Trochsa Arctosa(Pardosa Wadicosa))))。     

一株产纤维素酶细菌的筛选、鉴定及产酶条件优化

目的：筛选1株产纤维素酶的细菌。方法：通过对从腐烂朽木及其附近土壤中得到的样品进行富集培养、分离纯化得到16株纤维素分解菌,经刚果红染色鉴定和液体发酵培养后对其进行了菌种初步鉴定及产酶条件的初步优化。结果：获得1株纤维素酶分泌量较高的细菌LT3。结论：LT3为革兰氏阳性菌,菌体成杆状,经发酵优化培养后,较适产酶条件为甘蔗渣20g/L,pH7.0、30℃培养120h,CMC酶活为71.17U/mL,滤纸酶活为33.37U/mL。通过克隆其16S rDNA序列,对其进行系统进化分析,鉴定为蜡状芽孢杆菌。