Peter Schuchert

Medusozoan Phylogeny and Character Evolution Clarified by New Large and Small Subunit rDNA Data and an Assessment of the Utility of Phylogenetic Mixture Models

A newly compiled data set of nearly complete sequences of the large subunit of the nuclear ribosome (LSU or 28S) sampled from 31 diverse medusozoans greatly clarifies the phylogenetic history of Cnidaria. These data have substantial power to discern among many of the competing hypotheses of relationship derived from prior work. Moreover, LSU data provide strong support at key nodes that were equivocal based on other molecular markers. Combining LSU sequences with those of the small subunit of the nuclear ribosome (SSU or 18S), we present a detailed working hypothesis of medusozoan relationships and discuss character evolution within this diverse clade. Stauromedusae, comprising the benthic, so-called stalked jellyfish, appears to be the sister group of all other medusozoans, implying that the free-swimming medusa stage, the motor nerve net, and statocysts of ecto-endodermal origin are features derived within Medusozoa. Cubozoans, which have had uncertain phylogenetic affinities since the elucidation of their life cycles, form a clade-named Acraspeda-with the scyphozoan groups Coronatae, Rhizostomeae, and Semaeostomeae. The polyps of both cubozoans and hydrozoans appear to be secondarily simplified. Hydrozoa is comprised by two well-supported clades, Trachylina and Hydroidolina. The position of Limnomedusae within Trachylina indicates that the ancestral hydrozoan had a biphasic life cycle and that the medusa was formed via an entocodon. Recently hypothesized homologies between the entocodon and bilaterian mesoderm are therefore suspect. Laingiomedusae, which has often been viewed as a close ally of the trachyline group Narcomedusae, is instead shown to be unambiguously a member of Hydroidolina. The important model organisms of the Hydra species complex are part of a clade, Aplanulata, with other hydrozoans possessing direct development not involving a ciliated planula stage. Finally, applying phylogenetic mixture models to our data proved to be of little additional value over a more traditional phylogenetic approach involving explicit hypothesis testing and bootstrap analyses under multiple optimality criteria. [18S; 28S; Cubozoa; Hydrozoa; medusa; molecular systematics; polyp; Scyphozoa; Staurozoa.].

A specific DNA sequence is required for high frequency of recombination in the ade6 gene of fission yeast.

The point mutation M26 in the ade6 gene of Schizosaccharomyces pombe increases recombination frequency by an order of magnitude in comparison with other mutations in the same gene. The hypothesis is tested that this hot spot of recombination requires a specific nucleotide sequence at the M26 site. The DNA sequence is altered systematically by in vitro mutagenesis, and the resulting sequences are introduced into the ade6 gene in vivo by gene replacement. It results that any change of the heptanucleotide ATGACGT leads to loss of high frequency of recombination. Thus this oligonucleotide sequence is necessary for high frequency of recombination, but it seems not to be sufficient.

Phylogenetics of Hydroidolina (Hydrozoa: Cnidaria)

Hydroidolina is a group of hydrozoans that includes Anthoathecata, Leptothecata and Siphonophorae. Previous phylogenetic analyses show strong support for Hydroidolina monophyly, but the relationships between and within its subgroups remain uncertain. In an effort to further clarify hydroidolinan relationships, we performed phylogenetic analyses on 97 hydroidolinan taxa, using DNA sequences from partial mitochondrial 16S rDNA, nearly complete nuclear 18S rDNA and nearly complete nuclear 28S rDNA. Our findings are consistent with previous analyses that support monophyly of Siphonophorae and Leptothecata and do not support monophyly of Anthoathecata nor its component subgroups, Filifera and Capitata. Instead, within Anthoathecata, we find support for four separate filiferan clades and two separate capitate clades (Aplanulata and Capitata sensu stricto ). Our data however, lack any substantive support for discerning relationships between these eight distinct hydroidolinan clades.

SYNCHRONIZED MEIOSIS AND RECOMBINATION IN FISSION YEAST - OBSERVATIONS WITH PAT1-114 DIPLOID-CELLS

SummaryThe mutation pat1-114 has been used to synchronize meiosis in the fission yeast Schizosaccharomyces pombe. We have investigated several aspects of such synchronized meiotic cultures. In both pat1-114 and pat1+ diploids, meiotic landmark events are initiated at the same time after meiosis induction, but synchrony is much more pronounced in the pat1-114-driven meiosis. Commitment to recombination and to meiosis have been timed at 2 h after meiotic induction. Due to a seven-fold reduction of intragenic recombination frequency in the ade6 region of pat1-114 diploids, physical analysis of recombination has not been possible. We have distinguished three factors that influence intragenic recombination frequencies: temperature, azygotic versus zygotic meiosis, and the nature of the pat1 allele. Differences and similarities in the timing of meiotic landmarks in S. cerevisiae and S. pombe are discussed.

Molecular phylogenetics of Thecata (Hydrozoa, Cnidaria) reveals long-term maintenance of life history traits despite high frequency of recent character changes.

Two fundamental life cycle types are recognized among hydrozoan cnidarians, the benthic (generally colonial) polyp stage either producing pelagic sexual medusae or directly releasing gametes elaborated from an attached gonophore. The existence of intermediate forms, with polyps producing simple medusoids, has been classically considered compelling evidence in favor of phyletic gradualism. In order to gain insights about the evolution of hydrozoan life history traits, we inferred phylogenetic relationships of 142 species of Thecata (= Leptothecata, Leptomedusae), the most species-rich hydrozoan group, using 3 different ribosomal RNA markers (16S, 18S, and 28S). In conflict with morphology-derived classifications, most thecate species fell in 2 well-supported clades named here Statocysta and Macrocolonia. We inferred many independent medusa losses among Statocysta. Several instances of secondary regain of medusoids (but not of full medusa) from medusa-less ancestors were supported among Macrocolonia. Furthermore, life cycle character changes were significantly correlated with changes affecting colony shape. For both traits, changes did not reflect graded and progressive loss or gain of complexity. They were concentrated in recent branches, with intermediate character states being relatively short lived at a large evolutionary scale. This punctuational pattern supports the existence of 2 alternative stable evolutionary strategies: simple stolonal colonies with medusae (the ancestral strategy, seen in most Statocysta species) versus large complex colonies with fixed gonophores (the derived strategy, seen in most Macrocolonia species). Hypotheses of species selection are proposed to explain the apparent long-term stability of these life history traits despite a high frequency of character change. Notably, maintenance of the medusa across geological time in Statocysta might be due to higher extinction rates for species that have lost this dispersive stage.

Reconciling genealogical and morphological species in a worldwide study of the Family Hydractiniidae (Cnidaria, Hydrozoa)

The Hydractiniidae are a family of globally distributed marine hydrozoans (class Hydrozoa, phylum Cnidaria). Despite being one of the most well‐studied families of the Hydrozoa, their genus and species‐level taxonomy is unsettled and disputed. The taxonomic difficulties of the Hydractiniidae are due to many inadequate species descriptions, a paucity of available morphological characters, many cryptic species, and the often‐extreme plasticity seen when colonies of the same species are found at different stages of growth or different environmental conditions. This confusion over species identity is especially important because some species of the family Hydractiniidae are well‐established model organisms for a wide array of studies ranging from gene expression to developmental biology and colony growth. Here we report the species‐level implications of 226 mitochondrial large ribosomal subunit (16S) rDNA sequences from around the world and 52 nuclear DNA sequences (Elongation Factor 1α) with the intent to reconcile described morphospecies with genealogical lineages.

Phylogeny of the Plumularioidea (Hydrozoa, Leptothecata): evolution of colonial organisation and life cycle

The Plumularioidea (Cnidaria, Hydrozoa, Leptothecata) are the most species rich superfamily of the class Hydrozoa. They display a complex and diversified colonial organisation and their life cycle comprises either a reduced free‐living, pelagic generation (medusoid), alternating with the benthic colonial form or in most species, no pelagic generation. In order to understand the evolution of colonial and life cycle characters among Plumularioidea, we have reconstructed their phylogeny. Partial mitochondrial 16S rRNA sequences and 64 morphological characters were analysed separately and in combination. The morphological data included not only characters of the individual polyps and medusae, but also characters describing the organisation of colonies, for which we propose general principles applying to character coding in modular organisms. The phylogenetic analyses supported the monophyly of Plumularioidea and of the four plumularioid families (Aglaopheniidae, Halopterididae, Kirchenpaueriidae and Plumulariidae). Most genera were paraphyletic or polyphyletic. This study highlights multiple morphological simplifications of the colonial organisation during the evolution of Plumularioidea and the convergence of the defensive polyps — the dactylozooids — of Plumularioidea with those of others Leptothecata (Hydrodendron) or Anthoathecata (Hydractinia). Concerning the evolution of the life cycle, the phylogeny supports a provocative scenario, where the medusa was lost in an ancestor of the Plumularioidea, and then re‐acquired four times independently within this group, in the form of simple medusoids.

Life stage specific expression of a myosin heavy chain in the hydrozoan Podocoryne carnea

In order to study life stage specific gene-expression and also to investigate the molecular components of jellyfish muscle we cloned a tissue specifically expressed cDNA of the metagenetic hydrozoan Podocoryne carnea. A monoclonal antibody was made that reacts with the filaments of striated muscle of the medusa stage. The mAb detects a protein of 220 kDa in Western blots. The antibody was used to clone a cDNA from an expression library. Sequence analysis showed that the clone codes for the rod part of a myosing heavy chain (MHC). Surprisingly, the sequence is more similar to MHCs from striated muscle than to smooth muscle or non-muscle MHCs from either invertebrates or vertebrates. A particularly well conserved region, which may be correlated to a function in striated muscle, was found. Expression studies showed that the corresponding RNA is only present in the striated muscle cells of the medusa. During medusa development, the transcript became detectable at the same time as muscle differentiation becomes visible in the electron microscops.

High genetic diversity in the hydroid Plumularia setacea: A multitude of cryptic species or extensive population subdivision?

The marine hydroid Plumularia setacea has a near-cosmopolitan distribution. As in other sessile invertebrates with limited dispersal abilities, the wide distribution could also be a taxonomic artefact and the species might in fact be a complex of sibling species. To investigate this, a set of worldwide samples of P. setacea and several closely related species was examined using the mitochondrial markers 16S and COI, as well as the nuclear marker ITS. The results suggest an even higher degree of genetic diversity than expected. Almost all sampled regions had only private haplotypes and the resulting trees split into a multitude of geographically delimited lineages, this both for the mitochondrial and nuclear markers. In the framework of a genealogical species concept, these lineages would qualify as cryptic species. Using alternative species concepts, the results could be reconciled with traditional taxonomy by regarding P. setacea as a single species with an extensive population subdivision. A rapid molecular clock, limited dispersal abilities, and localized clonal propagation are likely the factors that explain the high but dispersed genetic diversity within this species.

Phylogenetics and evolution of Capitata (Cnidaria: Hydrozoa), and the systematics of Corynidae

Nawrocki, A. M., Schuchert, P. & Cartwright, P. (2009). Phylogenetics and evolution of Capitata (Cnidaria: Hydrozoa), and the systematics of Corynidae.—Zoologica Scripta, 39, 290–304.