Phylogeny of the Nemertodermatida (Acoelomorpha, Platyhelminthes). A cladistic analysis

The Nemertodermatida is a small group of worms, regarded as an order of the Platyhelminthes. The group is of special systematic interest because of its putative basal phylogenetic position in the Platyhelminthes. The phylogeny of the Nemertodermatida was estimated using paup 4.0 software for parsimony analysis. The analysis was based on 72 structural parsimony-informative characters totalling 184 different character states. All eight well described species of nemertodermatids were included in the ingroup. As outgroup were chosen species of the Acoela, Catenulida, Macrostomida and Xenoturbellida. A single most parsimonious tree was obtained with 140 steps and a consistency index (CI) of 0.80. The Nemertodermatida, Ascoparidae and Nemertodermatidae are shown as monophyletic taxa in the tree. The species Nemertoderma psammicola does not group with the other members of the genus Nemertoderma , hence criteria of phylogenetic taxonomy imply that N. psammicola should be renamed. A suggested new name is Sterreria psammicola gen. n .     

The muscular system of Nemertoderma westbladi and Meara stichopi (Nemertodermatida, Acoelomorpha)

Nemertodermatida is a small taxon of marine worm-like animals; its position in the tree of life is highly contested. The musculature of Nemertoderma westbladi and Meara stichopi is studied here in detail using fluorescent phalloidin and confocal microscopy. In both species, the musculature is composed of an outer layer of circular and an inner layer of longitudinal musculature, diagonal muscles form a distinct layer in N. westbladi, but in M. stichopi these fibres connect to both other layers. The supraterminally opening male pore and antrum are formed by invagination of the whole body-wall in both species, and the seminal vesicle is lined by a thin net of musculature only in full male maturity. Modifications of the ventral body-wall adjacent to the mouth are small and transient in N. westbladi including no extra musculature, whereas it consists of additional strong U-shaped musculature in M. stichopi. Myogenesis in N. westbladi is not finished in hatchlings and will be completed dorsally in juvenile specimens and ventrally in male mature ones, after the loss of the mouth. Musculature between the two species differs considerably and might give insights into the internal relationships of Nemertodermatida and might prove to be useful in studies investigating their phylogenetic position. More data of other species and developmental changes are needed.     

Evolution of body-wall musculature in the Platyhelminthes (Acoelomorpha, Catenulida, Rhabditophora)

In an effort to understand the phylogeny of the Platyhelminthes, the patterns of body-wall musculature of flatworms were studied using fluorescence microscopy and Alexa-488-labeled phalloidin. Species of the Catenulida have a simple orthogonal gridwork of longitudinal and circular muscles. Members of the Rhabditophora have the same gridwork of musculature, but also have diagonal muscles over their entire body. Although a few species of Acoelomorpha possessed a simple orthogonal grid of musculature, most species typically have distinctly different patterns of dorsal and ventral body-wall musculature that include sets of longitudinal, circular, U-shaped, and several kinds of diagonal muscles. Several distinct patterns of musculature were identified, including 8 patterns in 11 families of acoels. These patterns have proven to be useful in clarifying the phylogeny of the Acoelomorpha, particularly with regard to the higher acoels. Patterns of musculature as well as other morphological characters are used here for revisions of acoel systematics, including the return of Eumecynostomum sanguineum (Mecynostomidae) to the genus Aphanostoma (Convolutidae), the revision of the family Childiidae, and the formation of a new family, Actinoposthiidae.     

Molecular systematics of the Acoela (Acoelomorpha,Platyhelminthes) and its concordance with morphology

The phylogenetic relationships of the lower worm group Acoela were investigated using newly obtained nuclear 18S rDNA sequences from 16 acoels in combination with 16 acoel sequences available on GenBank from other laboratories. Parsimony and maximum likelihood analyses of the molecular data supported the concept that the Acoela is monophyletic; however, the gene tree produced by these analyses conflicts with the current taxonomic system for the Acoela in several family-level groupings. Most notable is the apparent polyphyly of the largest family of acoels, the Convolutidae. DNA analysis grouped together species of small-bodied convolutids in one clade, while large-bodied convolutids grouped in a separate clade with other large-bodied acoels. Despite such conflicts, the branching pattern in the gene tree is well supported by morphological characters of sperm and body-wall musculature.     

Ultrastructure of the epidermis of Meara stichopi (Platyhelminthes,Nemertodermatida) and associated extra-epidermal bacteria

This preliminary mechanistic model of normal swimming and phototactic behaviour in individual Daphnia was constructed using data and assumptions based on experiments and observations. Swimming under constant light intensity is characterized by short periods of upward movements alternating with equal periods of downward movements. Two oscillators are proposed that generate these phases in swimming. Unexpected shifts in depth, as observed in D. magna and D. longispina, are also present in the swimming of the computer daphnid and thus seem to be inherent to the underlying mechanism. As in real daphnids, during relative decreases in light intensity of low velocity, positive phototactic upward swimming is stepwise. With increasing velocity in the change in light, these steps disappear. When the model is triggered by a natural increase in light at dawn, a small downward movement results. Migration distance can be increased to commonly found depths of migrating Daphnia by the introduction of a fish exudate factor into the model, which enhances the phototactic response. Since attenuation of light in the water affects the phototactic swimming response, it also influences migration distance. The results of model calculations agree quite well with an empirical relationship between Secchi disc depth and amplitude of diel vertical migration in a number of lakes.     

Dismissal of Acoelomorpha: Acoela and Nemertodermatida are separate early bilaterian clades

We used new 18S and 28S rRNA sequences analysed with parsimony, maximum likelihood and Bayesian methods of phylogenetic reconstruction to show that Nemertodermatida, generally classified as the sister group of Acoela within the recently proposed Phylum Acoelomorpha, are a separate basal bilaterian lineage. We used several analytical approaches to control for possible long branch attraction (LBA) artefacts in our results. Parsimony and the model based phylogenetic reconstruction methods that incorporate 'corrections' for substitution rate heterogenities yielded concordant results. When putative long branch taxa were experimentally removed the resulting topologies were consistent with our total evidence analysis. Deletion of fast-evolving nucleotide sites decreased resolution and clade support, but did not support a topology conflicting with the total evidence analysis. Establishment of Acoela and Nemertodermatida as two early lineages facilitates reconstruction of ancestral bilaterian features. The ancestor of extant Bilateria was a small, benthic direct developer without coelom or a planktonic larval stage. The previously proposed Phylum Acoelomorpha is dismissed as paraphyletic.     

The brain of the Nemertodermatida (Platyhelminthes) as revealed by anti-5HT and anti-FMRFamide immunostainings

The taxa Nemertodermatida and Acoela have traditionally been considered closely related and classified as sister groups within the Acoelomorpha Ehlers 1984 (Platyhelminthes). Recent molecular investigations have questioned their respective position. In this study, the 5-HT and FMRFamide immunoreactivity (IR) in the nervous system of two nemertodermatids, Nemertoderma westbladi and Meara stichopi, is described. The 5-HT immunoreactive pattern differs in the two nemertodermatids studied. In M. stichopi, two loose longitudinal bundles of 5-HT-immunoreactive fibres and an basi-epidermal nerve net were observed. In N. westbladi the 5-HT-IR shows a ring-shaped commissural structure, different from the commissural brain of acoels. In both nemertodermatids, FMRFamide immunoreactive nerve fibres followed the 5-HT-immunoreactive fibres. It is demonstrated that the Nemertodermatida have neither a 'commissural brain' structure similar to that of the Acoela, nor a 'true', ganglionic brain and orthogon, typical for other Platyhelminthes. The question of the plesiomorphic or apomorphic nature of the nervous system in Nemertodermatida cannot yet be answered. The neuroanatomy of the studied worms provides no synapomorphy supporting the taxon Acoelomorpha.     

The Nemertodermatida are basal bilaterians and not members of the Platyhelminthes

Recent hypotheses on metazoan phylogeny have recognized three main clades of bilaterian animals: Deuterostomia, Ecdysozoa and Lophotrochozoa. The acoelomate and 'pseudocoelomate' metazoans, including the Platyhelminthes, long considered basal bilaterians, have been referred to positions within these clades by many authors. However, a recent study based on ribosomal DNA placed the flatworm group Acoela as the sister group of all other extant bilaterian lineages. Unexpectedly, the nemertodermatid flatworms, usually considered the sister group of the Acoela together forming the Acoelomorpha, were grouped separately from the Acoela with the rest of the Platyhelminthes (the Rhabditophora) within the Lophotrochozoa. To re-evaluate and clarify the phylogenetic position of the Nemertodermatida, new sequence data from 18S ribosomal DNA and mitochondrial genes of nemertodermatid and other bilaterian species were analysed with parsimony and maximum likelihood methods. The analyses strongly support a basal position within the Bilateria for the Nemertodermatida as a sister group to all other bilaterian taxa except the Acoela. Despite the basal position of both Nemertodermatida and Acoela, the clade Acoelomorpha was not retrieved. These results imply that the last common ancestor of bilaterian metazoans was a small, benthic, direct developer without segments, coelomic cavities, nephrida or a true brain. The name Nephrozoa is proposed for the ancestor of all bilaterians excluding the Nemertodermatida and the Acoela, and its descendants.     

The epidermal ciliary rootlets of Xenoturbella bocki (Xenoturbellida) revisited: new support for a possible kinship with the Acoelomorpha (Platyhelminthes)

Ultrastructural studies of epidermal locomotory ciliary structures of Xenoturbella bocki , i.e. the basal part of the axaneme, the basal body and the ciliary rootlets, have revealed characters supporting the hypothesis of a close relationship between Xenoturbella and the Acoelomorpha. Some of the most prominent of these possibly synapomorphic characters are a cup-shaped structure at the base of the axoneme, a proximally hollow rootlet arising on the anterior face of the basal body and bundles of microtubules extending from the basal foot to the main rootlets in the next hind row. Other views in the recent literature places Xenoturbella as closely related to the Mollusca or to the stem species of the Bilateria. As such, the phylogenetic affinities of Xenoturbella remain highly debateable. A review of recent reports on the subject is given here, together with a discussion of combinations of current phylogeny hypotheses.     

Mitochondrial genome data support the basal position of Acoelomorpha and the polyphyly of the Platyhelminthes

We determined 9.7, 5.2, and 6.8 kb, respectively, of the mitochondrial genomes of the acoel Paratomella rubra, the nemertodermatid Nemertoderma westbladi, and the free-living rhabditophoran platyhelminth Microstomum lineare. The identified gene arrangements are unique among metazoans, including each other, sharing no more than one or two single gene boundaries with a few distantly related taxa. Phylogenetic analysis of the amino acid sequences inferred from the sequenced genes confirms that the acoelomorph flatworms (acoels+nemertodermatids) do not belong to the Platyhelminthes, but are, instead, the most basal extant bilaterian group. Therefore, the Platyhelminthes, as traditionally constituted, is a polyphyletic phylum.     

达氏科(扁形动物门,单肠目)涡虫二新种

本文记述淡水涡虫类单肠目Rhabdocoela达氏科Dalyelliidae小达氏涡虫属Microdalyellia一新种,广东小达氏涡虫Microdelyellia guangdongensis Wang et Rong,sp.nov.,模式标本由卢彦宏于2013年6月9日采自广东省深圳市石岩水库(22°40'58″N,113°53'42″E);杰氏涡虫属Gieysztoria一新种,七星岩杰氏涡虫Gieysztoria qixingyanensis Wang et Rong,sp.nov.,模式标本由刘旭坤于2014年8月6日采自广东省肇庆市七星岩风景区(23°04'32″N,112°28'46″E)。对2个涡虫新种的形态特征作了详细描述,并分别与小达氏涡虫属和杰氏涡虫属近似物种进行了比较。所有标本保存于中国科学院国家动物博物馆。     

Two Intracellular Symbiotic Bacteria from the Mulberry Psyllid Anomoneura mori (Insecta, Homoptera)

We characterized the intracellular symbiotic bacteria of the mulberry psyllid Anomoneura mori by performing a molecular phylogenetic analysis combined with in situ hybridization. In its abdomen, the psyllid has a large, yellow, bilobed mycetome (or bacteriome) which consists of many round uninucleated mycetocytes (or bacteriocytes) enclosing syncytial tissue. The mycetocytes and syncytium harbor specific intracellular bacteria, the X-symbionts and Y-symbionts, respectively. Almost the entire length of the bacterial 16S ribosomal DNA (rDNA) was amplified and cloned from the whole DNA of A. mori, and two clones, the A-type and B-type clones, were identified by restriction fragment length polymorphism analysis. In situ hybridization with specific oligonucleotide probes demonstrated that the A-type and B-type 16S rDNAs were derived from the X-symbionts and Y-symbionts, respectively. Molecular phylogenetic analyses of the 16S rDNA sequences showed that these symbionts belong to distinct lineages in the γ subdivision of the Proteobacteria. No 16S rDNA sequences in the databases were closely related to the 16S rDNA sequences of the X- and Y-symbionts. However, the sequences that were relatively closely related to them were the sequences of endosymbionts of other insects. The nucleotide compositions of the 16S rDNAs of the X- and Y-symbionts were highly AT biased, and the sequence of the X-symbiont was the most AT-rich bacterial 16S rDNA sequence reported so far.     

昆虫病原线虫的共生细菌

昆虫病原线虫与其共生细菌二者互惠共生 :共生细菌需要昆虫病原线虫作为载体以寄生寄主昆虫并做为自己的营养来源 ,而昆虫病原线则需要依靠共生细菌来杀死昆虫。综述了共生细菌的病原作用、抗菌作用与杀虫作用 ,评述了共生细菌的基因工程进展 ,讨论了昆虫共生细菌在昆虫病原线虫致病性的作用。     

大口涡虫属中国一新纪录(扁形动物门,大口虫目)及其分类性状

报道了大口虫目大口虫科中国一新纪录种,即帆大口涡虫Macrostomum saifunicum Nasonov,1929,详细研究了该涡虫的个体发育与角质阴茎发育的关系.结果 表明:1) 帆大口涡虫角质阴茎端部结构在个体发育的第9 d 一次性成型,终身不变,是一个稳定的关键分类性状; 2) 个体发育至第20 d 后,角质阴茎长度达140 μm,第40 d后个体进入衰老期,其角质阴茎长度超过160 μm,指出角质阴茎的长度不是一个稳定的分类性状; 3) 根据作者多年的野外采集,发现北京、安徽、湖南、江西、广东均有帆大口涡虫分布,指出该物种是中国常见的分布较广的物种,并提示大口涡虫是一种在教学和科学研究方面理想的实验动物.     

Comparative morphology of the bursal nozzles in acoels (Acoela, Acoelomorpha)

Systematics of the Acoela is particularly difficult because of the paucity of readily discernible morphological features. In other soft-bodied worms, sclerotized structures, such as copulatory stylets, provide important characters that can be seen in whole mounts, but acoels generally lack such features. Among the few sclerotized structures in acoels are bursal nozzles-tubiform outlets on the seminal bursae that are believed to be conduits (spermatic ducts) through which allosperm are transported to the oocytes. Early classifications of the Acoela used features of the female reproductive system, including bursal nozzles, for distinguishing major groups, but the current system essentially ignores them as too plastic to provide higher-level distinctions. We used confocal and electron microscopy to further characterize bursal nozzles in five acoel species, and found all composed of actin-reinforced extensions of stacked, flat mesenchymal cells. In Notocelis gullmarensis, Aphanostoma bruscai, and Daku woorimensis, the nozzle is a stiffened region of the same cells forming the wall of the bursa. By contrast, in Wulguru cuspidata cells forming the nozzle are distinct from those of the bursa. The so-called bursal cap of A. bruscai and D. woorimensis has small sclerotized disjunct units within it, also composed of stacked, flat, actin-reinforced cells. The nozzle of W. cuspidata, prominent like that of other convolutid acoels, is relatively complex, its actin-reinforced cells sandwiched with secretory cells and its base bearing a "sorting apparatus" of egg-shaped cells that send narrow processes inside the spermatic duct. Cases of sperm inside the nozzle corroborate its assumed role in reproduction. Whereas most nozzles sit at the end of the bursa facing the ovary, in species of Pseudmecynostomum and purportedly in a few other acoels, they sit between the female pore and the bursa, constituting what we call a vaginal nozzle. All bursal nozzles of acoels show a common ground pattern indicating common ancestry, but certain features discerned through electron and confocal microscopy show promise of providing synapomorphies for grouping some species.     

Hox genes from the Polystomatidae (Platyhelminthes, Monogenea)

Hox genes form a multigenic family that play a fundamental role during the early stages of development. They are organised in a single cluster and share a 60 amino acid conserved sequence that corresponds to the DNA binding domain, i.e. the homeodomain. Sequence conservation in this region has allowed investigators to explore Hox diversity in the metazoan lineages. Within parasitic flatworms only homeobox sequences of parasite species from the Cestoda and Digenea have been reported. In the present study we surveyed species of the Polyopisthocotylea (Monogenea) in order to clarify Hox identification and diversification processes in the neodermatan lineage. From cloning of degenerative PCR products of the central region of the homeobox, we report one ParaHox and 25 new Hox sequences from 10 species of the Polystomatidae and one species of the Diclidophoridae, which extend Hox gene diversity from 46 to 72 within Neodermata. Hox sequences from the Polyopisthocotylea were annotated and classified from sequence alignments and Bayesian inferences of 178 Hox, ParaHox and related gene families recovered from all available parasitic platyhelminths and other bilaterian taxa. Our results are discussed in the light of the recent Hox evolutionary schemes. They may provide new perspectives to study the transition from turbellarians to parasitic flatworms with complex life-cycles and outline the first steps for evolutionary developmental biological approaches within platyhelminth parasites.     

中国伞形科矮泽芹属植物叶表皮形态特征的研究

利用光学显微镜、扫描电镜对伞形科矮泽芹属8种植物叶表皮形态进行观察与研究。结果表明:(1)矮泽芹属8种植物上下表皮细胞均为不规则形或规则多边形,垂周壁为近平直状或波状,上表皮细胞长宽比在1.3~2.4之间,下表皮细胞长宽比在1.5~2.5之间;在近轴面,有细叶矮泽芹、聂拉木矮泽芹和绿花矮泽芹3种植物没有气孔器的存在,其余物种气孔密度在20~74个/mm2之间,气孔指数为6.0%~17.7%;在远轴面,所有物种都具有丰富的气孔器,气孔密度为100~183个/mm2,气孔指数为16.1%~23.6%。(2)聚类分析结果显示,矮泽芹、大苞矮泽芹、粗棱矮泽芹为类群Ⅰ,鹤庆矮泽芹为类群Ⅱ,聂拉木矮泽芹、细叶矮泽芹、绿花矮泽芹为类群Ⅲ,松潘矮泽芹则单独聚为类群Ⅳ;聚类分析结果大体上支持形态学分类的结果。(3)叶表皮形态特征对于区分矮泽芹属不同物种具有十分重要的分类学价值。     

Ultrastructure of the pharynx of Prorhynchus (Platyhelminthes, Lecithoepitheliata)

The pharynx variabilis of Prorhynchus is strongly muscular, with a small pharyngeal fold and a thin surrounding sheath. There is one row of inner longitudinal musclcs, up to six rows of inner circular muscles, many radial muscles, one row of outer circular and one row of outer longitudinal muscles, with no sphincter muscle groups. Three kinds of secretion, produced in a cluster of gland cell bodies posterior to the pharynx, enter the pharynx wall. They travel anteriorly in ducts and two kinds unite in a common duct just prior to discharging into the anterior region of the pharynx lumen. The perikarya of lumen epithelial cells lie within the pharynx musculature and, at the anterior and posterior margins of the pharynx, external to the pharynx. Bundles of ciliated receptors are numerous at the anterior and posterior constrictions. Similarities in the ultrastructure of flame bulbs of Rhabdocoela and Lecithoepitheliata suggest a relationship between these groups. However, the usefulness of pharynx ultrastructure for platyhelminth phylogeny cannot be assessed until complete ultrastructural studies of various groups of Rhabdocoela have been made.     

中国涡虫新纪录科微口涡虫属一新种(大口虫目,微口虫科)及其分子系统学分析

本文报道中国涡虫新纪录科,中国微口涡虫,新种Microstomum sinensis n.sp.。标本采自中国广东省深圳市南山区蛇口码头海边。该新种具有横裂殖和有性生殖个体。所有个体均具雄性生殖器官,其交配刺骨质,整体呈钩状,全长60~75μm,基部呈漏斗状,从基部到末端刺管直径逐渐缩小。交配刺后1/3弯曲位于基部所在平面之外,弯曲程度接近半圆,末端开口呈尖叶状。另对其16S r DNA和28S r DNA进行了分子系统学分析,确定了其分类地位。