六足动物分子系统学研究进展

对近期国内外六足总纲动物的原尾纲、弹尾纲、双尾纲和昆虫纲在种群遗传变异及进化、种及种下阶元的分类鉴定、种上阶元的系统发育分析等分子系统学方面的研究进展进行了综述。多基因的联合分子数据研究日益增加。随着分子技术的日益推广,不同类型的基因序列甚至全基因组的联合使用将引导分子系统学走向辉煌的未来。     

六足亚门内颚纲问题浅析

依据近年来在六足动物系统学研究中取得的最新成果,对一些动物学教材中有关六足动物中内颚纲的有效性、进化地位等问题进行浅析,提供原尾纲、弹尾纲和双尾纲的形态、习性和代表物种的相关更新资料,供生物学专业的师生参考.     

跳虫系统进化的研究进展

跳虫是弹尾纲（Collembola）的俗称,在所有六足动物中化石年代最早,因此跳虫是六足动物起源及进化研究中非常重要的类群。跳虫的起源、分类地位和系统关系等问题,对于阐明六足动物甚至节肢动物各大类群的系统关系非常关键,日益成为相关学者关注和争论的焦点。本文就跳虫形态学和分子系统学方面的研究工作进行了综述。     

六足总纲系统发育研究进展与新分类系统

简要综述了昆虫分纲、分目的历史变化,包括昆虫分目多少的变化,昆虫是纲级还是总纲级阶元的变化,昆虫各目分类地位系统排列的变化以及六足总纲系统发育研究进展。根据近10年来形态特征与分子测序数据相结合的系统发育研究,整理出六足总纲与系统发育支序分析相一致的分类系统,对昆虫35目的分类运用了10个分类阶元。在此基础上,删减次要分类阶元,提出简明分类系统,既反映每个高级分类单元的单系性,明晰各目的共祖近度,又减少了分类阶元层次,方便各分类单元的识别与鉴定。六足总纲Hexapoda分为4纲:原尾纲Protura(包括蚖目Acerentomata、华蚖目Sinentomata、古蚖目Eosentomata),弹尾纲Collembola(包括弹尾目Collembola),双尾纲Diplura(包括双尾目Diplura),昆虫纲Insecta。昆虫纲分为单髁亚纲SubclassMonocondylia(包括石蛃目Archaeognatha)与双髁亚纲SubclassDicondylia。双髁亚纲分为衣鱼部DivisionZygentoma(包括衣鱼目Zygentoma)与有翅部DivisionPterygota。有翅部分为10个总目、27目。     

综合纲系统学研究进展

综合纲是形态十分特殊的多足动物类群,其系统发育地位一直备受争议。目前,综合纲的分类学和系统发育研究相较其他节肢动物类群欠缺,全世界仅记录204种,且主要集中在欧洲、美洲和大洋洲,亚洲和非洲地区的研究十分匮乏。综合纲1目2科13属的分类体系较为完备并被广泛采用,但其内部幺蚰科和幺蚣科之间进化关系尚不明确。综合纲与少足纲、倍足纲共同组成前殖孔类的系统发育假说得到了形态学及分子序列数据等方面的支持,但意见分歧依然存在。中国综合纲动物的系统学研究较为薄弱,目前仅有零星的报道。现从经典分类学和系统发育学两个方面对综合纲系统学研究的最新进展进行综述,以期推动中国综合纲动物系统学研究的开展。     

从弹尾纲和原尾纲的亲缘关系质疑缺尾纲（=近昆虫纲）的有效性

鉴于弹尾目（跳虫）和原尾目的尾部都没有尾须（cerci）,Brner于1910年就把这两类归并为一个类群缺尾纲（Ellipura）,这一分类阶元长期被许多昆虫学家沿用至今。Kukalová-Peck(1987 )在讨论化石双尾虫（?）附肢的总体结构（ground plan）时,认为跳虫和原尾虫的腹部侧板更原始,附肢无转节,将二者归纳成近昆虫纲（Parainsecta）。但是从形态特征、内部结构、比较精子学、变态类型和胚后发育等的特点以及线粒体DNA和核糖体DNA的测序分析结果,显示弹尾纲与原尾纲之间存在诸多重要差异,不具备较为密切的亲缘关系,我们不支持（弹尾纲+原尾纲）组成缺尾纲或近昆虫纲。据此建议取消缺尾纲（=近昆虫纲）这一分类阶元。     

节肢动物的分类和演化

节肢动物门是动物界中最大的1个门。按新的分类系统,本门分三叶虫亚门(已灭绝)、螯肢亚门、甲壳亚门、六足亚门和多足亚门等5亚门。六足亚门相当于以前分类系统中的昆虫纲(广义的), 是最重要的一类节肢动物;此亚门分原尾纲、弹尾纲、双尾纲和昆虫纲(狭义的)等4纲;昆虫纲分3亚纲30目,包括了前昆虫纲“有翅亚纲”中的各目。为便于读者了解新、旧系统的异同,文中列举前人的代表性系统加以对照。同时,对学术界关于节肢动物的起源和演化的一些新观点予以必要的说明,对现时流行的“泛节肢动物”、“泛甲壳动物”等概念作了简要的介绍。     

有关节肢动物系统发育若干问题的研究进展

近年来对节肢动物系统发育问题的认识有了相当大的变化,尤其是支序分析研究方法的使用,以及来自分子系统学和分子发育生物学的证据,成为继比较形态学之后有力的研究手段,近年来的研究显示线虫,缓步类和有爪类是节肢动物的近缘类群,而非以前绝大多数学者所认为的软体动物的环节动物,对于节肢动物门的单系性及其内部类群间关系的问题,目前倾向于认为（1）节肢动物为单系群;（2）由甲壳类,六足类和多足类联合形成单系群－具颚类;（3）与六足类分子系统学两方面研究的支持;对第3点的争议虽较大,但就论据而言确为优势观点。     

一种改进的小型节肢动物无形态损伤的DNA提取方法

本文介绍了一种改进的小型节肢动物无形态损伤的DNA提取方法,并在双尾纲、原尾纲和弹尾纲中进行了实验验证。结果表明,该方法可以高效的提取三类小型节肢动物的DNA,并用于扩增目标基因序列,凭证标本的回收质量高,有助于进一步的分类鉴定。该方法有望对螨、蚜虫、介壳虫、蚤等其他小型节肢动物的分子鉴定提供方便。     

双尾虫系统进化的初步探讨

双尾虫系统发生问题目前血受学者的关注。作为六足总纲中内,外颚亚纲的过渡类群,双尾虫是否单系性及其系统地位的确定是争论的焦点,也是解决六足总纲高级阶元系统关系的一个关键。文中综述了多年来双尾虫形态学和解剖学方面研究所引发的争议和达成的共识,并对近几年该方面分子系统学的工作进行了总结和初步探讨。     

Testing for misleading effects in the phylogenetic reconstruction of ancient lineages of hexapods: influence of character dependence and character choice in analyses of 28S rRNA sequences

The present analyses employ the almost complete sequence of the 28S rRNA gene to investigate phylogenetic relationships among Pancrustacea, placing special emphasis on the position of basal hexapod lineages. This study utilizes a greater number of characters and taxa of Protura, Collembola and Diplura than previous analyses to focus on conflicts in the reconstruction of the early steps in hexapod evolution. Phylogenetic trees are mainly based on Bayesian approaches, but likewise include analyses with Maximum Likelihood and Maximum Parsimony. Different analyses, including the application of a mixed DNA/RNA substitution model, were performed to narrow possible misleading effects of non-stationarity of nucleotide frequencies, saturation and character independence down to a minimum. This is the first time that a mixed DNA/RNA model is applied to analyse 28S rRNA sequences of basal hexapods. All methods yielded strong support for the monophyly of Collembola, Diplura, Dicondylia and Insecta s.str. , as well as for a cluster composed of Diplura and Protura ('Nonoculata-hypothesis'). However, the last cluster may be an artifact caused by a shared GC bias of the 28S sequences between these orders, in combination with a long branch effect. The instability of the position of the 'Nonoculata' within Pancrustacea further bears out the misleading effect of non-stationarity of nucleotide frequencies. Protura and Diplura either form the sister-group to Collembola (Entognatha) or cluster with branchiopod crustaceans. Overall, the phylogenetic signal of the complete sequences of the 28S rRNA gene favours monophyly of Hexapoda over paraphyly. However, further corroboration from independent data is needed to rule out the competing hypothesis of mutually paraphyletic Crustacea and Hexapoda.     

MicroRNA evolution provides new evidence for a close relationship of Diplura to Insecta

The phylogenetic interrelationships among four hexapod lineages (Protura, Collembola, Diplura and Insecta) are pivotal to understanding the origin of insects and the early diversification of Hexapoda, but they have been difficult to clarify based on the available data. In this study, we identified 91 conserved microRNA (miRNA) families from 36 panarthropod taxa, including seven newly sequenced non-insect hexapods. We found major clade differentiation accompanied by the origin of novel miRNA families, and most miRNA clusters are conserved with a high degree of microsynteny. Importantly, we were able to identify two miRNA families unique to Hexapoda, and four miRNA families and a miRNA cluster that exist exclusively in Diplura and Insecta, suggesting a close relationship between Diplura and Insecta as well as the monophyly of Hexapoda. Combined with a phylogenetic analysis based on the presence/absence matrix of miRNA families, our study demonstrates the effectiveness of miRNA in resolving deep phylogenetic problems.     

Is Ellipura monophyletic? A combined analysis of basal hexapod relationships with emphasis on the origin of insects

Hexapoda includes 33 commonly recognized orders, most of them insects. Ongoing controversy concerns the grouping of Protura and Collembola as a taxon Ellipura, the monophyly of Diplura, a single or multiple origins of entognathy, and the monophyly or paraphyly of the silverfish (Lepidotrichidae and Zygentoma s.s.) with respect to other dicondylous insects. Here we analyze relationships among basal hexapod orders via a cladistic analysis of sequence data for five molecular markers and 189 morphological characters in a simultaneous analysis framework using myriapod and crustacean outgroups. Using a sensitivity analysis approach and testing for stability, the most congruent parameters resolve Tricholepidion as sister group to the remaining Dicondylia, whereas most suboptimal parameter sets group Tricholepidion with Zygentoma. Stable hypotheses include the monophyly of Diplura, and a sister group relationship between Diplura and Protura, contradicting the Ellipura hypothesis. Hexapod monophyly is contradicted by an alliance between Collembola, Crustacea and Ectognatha (i.e., exclusive of Diplura and Protura) in molecular and combined analyses.     

Ribosomal DNA gene and phylogenetic relationships of Diplura and lower Hexapods

HennigdividedInsectas.lat.(=Hexapoda)intotwowelldefinedtaxonomicgroups:Entog-nathaandEctognatha[1].ThemostdistinctivecharacterofEntognatha(includingProtura,Col-lembolaandDiplura)liesintheenclosedmouthpartscondition,whereasthepresenceofexposedmouthpartsisthemainfeatureofEctognatha(includingMicrocoryphia,Zygentomaandtheptery-goteinsects).ControversiesaboutthephylogeneticrelationshipsofhightaxaEntognathahavekeptgrowinginrecentyears,withthemonophylyofDipluraandthephylogeneticpositionsofDiplur…     

Ribosomal DNA gene and phylogenetic relationships of Diplura and lower Hexapods

The monophyly of Diplura and its phylogenetic relationship with other hexapods are important for understanding the phylogeny of Hexapoda. The complete 18SrRNAgene and partial 28SrRNA gene (D3-D5 region) from 2 dipluran species (Campodeidae and Japygidae), 2 proturan species, 3 collembolan species, and 1 locust species were sequenced. Combining related sequences in GenBank, phylogenetic trees of Hexapoda were constructed by MP method using a crustaceanArtemia salina as an outgroup. The results indicated that: (i) the integrated data of 18SrDNA and 28SrDNA could provide better phylogenetic information, which well supported the monophyly of Diplura; (ii) Diplura had a close phylogenetic relationship to Protura with high bootstrap support.     

Brain organization in Collembola (springtails)

Arthropoda is comprised of four major taxa: Hexapoda, Crustacea, Myriapoda and Chelicerata. Although this classification is widely accepted, there is still some debate about the internal relationships of these groups. In particular, the phylogenetic position of Collembola remains enigmatic. Some molecular studies place Collembola into a close relationship to Protura and Diplura within the monophyletic Hexapoda, but this placement is not universally accepted, as Collembola is also regarded as either the sister group to Branchiopoda (a crustacean taxon) or to Pancrustacea (crustaceans + hexapods). To contribute to the current debate on the phylogenetic position of Collembola, we examined the brains in three collembolan species: Folsomia candida, Protaphorura armata and Tetrodontophora bielanensis, using antennal backfills, series of semi-thin sections, and immunostaining technique with several antisera, in conjunction with confocal laser scanning microscopy and three-dimensional reconstructions. We identified several neuroanatomical structures in the collembolan brain, including a fan-shaped central body showing a columnar organization, a protocerebral bridge, one pair of antennal lobes with 20-30 spheroidal glomeruli each, and a structure, which we interpret as a simply organized mushroom body. The results of our neuroanatomical study are consistent with the phylogenetic position of Collembola within the Hexapoda and do not contradict the hypothesis of a close relationship of Collembola, Protura and Diplura.     

The phylogenetic positions of three Basal-hexapod groups (protura, diplura, and collembola) based on ribosomal RNA gene sequences

This study combined complete 18S with partial 28S ribosomal RNA gene sequences ( approximately 2,000 nt in total) to investigate the relations of basal hexapods. Ten species of Protura, 12 of Diplura, and 10 of Collembola (representing all subgroups of these three clades) were sequenced, along with 5 true insects and 8 other arthropods, which served as out-groups. Trees were constructed with maximum parsimony, maximum likelihood, Bayesian analysis, and minimum-evolution analysis of LogDet-transformed distances. All methods yielded strong support for a clade of Protura plus Diplura, here named Nonoculata, and for monophyly of the Diplura. Parametric-bootstrapping analysis showed our data to be inconsistent with previous hypotheses (P < 0.01) that joined Protura with Collembola (Ellipura), that said Diplura are sister to true insects or are diphyletic, and that said Collembola are not hexapods. That is, our data are consistent with hexapod monophyly and Collembola grouped weakly with "Protura + Diplura" under most analytical conditions. As a caveat to the above conclusions, the sequences showed nonstationarity of nucleotide frequencies across taxa, so the CG-rich sequences of the diplurans and proturans may have grouped together artifactually; however, the fact that the LogDet method supported this group lessens this possibility. Within the basal hexapod groups, where nucleotide frequencies were stationary, traditional taxonomic subgroups generally were recovered: i.e., within Protura, the Eosentomata and Acerentomata (but Sinentomata was not monophyletic); within Collembola, the Arthropleona, Poduromorpha, and Entomobryomorpha (but Symphypleona was polyphyletic); and in Diplura, the most complete data set (> 2,100 nt) showed monophyly of Campodeoidea and of Japygoidea, and most methods united Projapygoidea with Japygoidea.     

The phylogenetic interrelationships of the higher taxa of apterygote hexapods

The phylogeny of the basal hexapods, the so-called apterygote insects, was studied using parsimony analysis procedures. Most analyses took into account 47 characters mainly based on external morphology, and 19 taxa including 14 apterygote representatives, 3 pterygotes and also 2 distantly related myriapods were used as outgroups. The binary and multistate characters are discussed in detail and treated as unordered and equally weighted. Other analyses were performed using a second data set in which 28 characters, based on internal anatomy and already used in a previous work ( Bitsch & Bitsch 1998 ), were added to the first data set. This second matrix was restricted to 12 terminal taxa, the same as those of our previous work. The results of the different analyses are generally congruent. They strongly support the monophyly of several orders (Protura, Collembola, Archaeognatha) and of two groupings (Ectognatha, Dicondylia). Three other assemblages (Ellipura, Diplura, Entognatha) appear as parsimonious phylogenetic hypotheses, but they are never supported by the cladistical analyses and are based on a very small number of autapomorphies; so, the monophyly of each of them is not firmly established. Archaeognatha appears as the sister group of the Dicondylia. The three unresolved representatives of the Zygentoma are found as the sister group of the Pterygota. The results are discussed in the light of current concepts in hexapod phylogeny.     

Towards an 18S phylogeny of hexapods: accounting for group-specific character covariance in optimized mixed nucleotide/doublet models

The phylogenetic diversification of Hexapoda is still not fully understood. Morphological and molecular analyses have resulted in partly contradicting hypotheses. In molecular analyses, 18S sequences are the most frequently employed, but it appears that 18S sequences do not contain enough phylogenetic signals to resolve basal relationships of hexapod lineages. Until recently, character interdependence in these data has never been treated seriously, though possibly accounting for the occurrence of biased results. However, software packages are readily available which can incorporate information on character interdependence within a Bayesian approach. Accounting for character covariation derived from a hexapod consensus secondary structure model and applying mixed DNA/RNA substitution models, our Bayesian analysis of 321 hexapod sequences yielded a partly robust tree that depicts many hexapod relationships congruent with morphological considerations. It appears that the application of mixed DNA/RNA models removes many of the anomalies seen in previous studies. We focus on basal hexapod relationships for which unambiguous results are missing. In particular, the strong support for a “Chiastomyaria” clade (Ephemeroptera+Neoptera) obtained in Kjer's [2004. Aligned 18S and insect phylogeny. Syst. Biol. 53, 1–9] study of 18S sequences could not be confirmed by our analysis. The hexapod tree can be rooted with monophyletic Entognatha but not with a clade Ellipura (Collembola+Protura). Compared to previously published contributions, accounting for character interdependence in analyses of rRNA data presents an improvement of phylogenetic resolution. We suggest that an integration of explicit clade-specific rRNA structural refinements is not only possible but an important step in the optimization of substitution models dealing with rRNA data.     

Phylogenetic relationships of basal hexapods among the mandibulate arthropods: a cladistic analysis based on comparative morphological characters

In this paper we propose a reappraisal of the relationships between the basal hexapod lineages (the former 'apterygote' insects) and the other major groups of mandibulate arthropods. It results from a cladistic analysis including 72 characters based on external morphology, internal anatomy and development. Detailed comments are provided on the various characters used and the scoring of their states. The 35 terminal taxa include 12 hexapods (9 of which are basal 'apterygote' representatives), 7 myriapods, 13 crustaceans, and 3 chelicerates taken as outgroups. The results of our analyses are discussed in detail for each of the taxonomic groupings, and compared with those recently obtained by other authors using different approaches based on morphological, palaeontological, developmental or molecular sequence data. Our results support the monophyly of the Mandibulata, Crustacea, Atelocerata (Tracheata) and Hexapoda, but the assemblage of Myriapoda appears poorly supported. A close relationship between Crustacea and Hexapoda, as hypothesized by several authors, is not found in any of our analyses. Within Hexapoda, the Protura and the Collembola appear as independent clades, whereas the two unresolved dipluran taxa are grouped with the monophyletic Ectognatha (Archaeognatha, Zygentoma and Pterygota).