Annette Klussmann-Kolb

Opisthobranchia (Mollusca, Gastropoda) - more than just slimy slugs. Shell reduction and its implications on defence and foraging.

BackgroundIn general shell-less slugs are considered to be slimy animals with a rather dull appearance and a pest to garden plants. But marine slugs usually are beautifully coloured animals belonging to the less-known Opisthobranchia. They are characterized by a large array of interesting biological phenomena, usually related to foraging and/or defence. In this paper our knowledge of shell reduction, correlated with the evolution of different defensive and foraging strategies is reviewed, and new results on histology of different glandular systems are included.ResultsBased on a phylogeny obtained by morphological and histological data, the parallel reduction of the shell within the different groups is outlined. Major food sources are given and glandular structures are described as possible defensive structures in the external epithelia, and as internal glands.ConclusionAccording to phylogenetic analyses, the reduction of the shell correlates with the evolution of defensive strategies. Many different kinds of defence structures, like cleptocnides, mantle dermal formations (MDFs), and acid glands, are only present in shell-less slugs. In several cases, it is not clear whether the defensive devices were a prerequisite for the reduction of the shell, or reduction occurred before. Reduction of the shell and acquisition of different defensive structures had an implication on exploration of new food sources and therefore likely enhanced adaptive radiation of several groups.

Transcriptomic Evidence That Longevity of Acquired Plastids in the Photosynthetic Slugs Elysia timida and Plakobranchus ocellatus Does Not Entail Lateral Transfer of Algal Nuclear Genes

Sacoglossan sea slugs are unique in the animal kingdom in that they sequester and maintain active plastids that they acquire from the siphonaceous algae upon which they feed, making the animals photosynthetic. Although most sacoglossan species digest their freshly ingested plastids within hours, four species from the family Plakobranchidae retain their stolen plastids (kleptoplasts) in a photosynthetically active state on timescales of weeks to months. The molecular basis of plastid maintenance within the cytosol of digestive gland cells in these photosynthetic metazoans is yet unknown but is widely thought to involve gene transfer from the algal food source to the slugs based upon previous investigations of single genes. Indeed, normal plastid development requires hundreds of nuclear-encoded proteins, with protein turnover in photosystem II in particular known to be rapid under various conditions. Moreover, only algal plastids, not the algal nuclei, are sequestered by the animals during feeding. If algal nuclear genes are transferred to the animal either during feeding or in the germ line, and if they are expressed, then they should be readily detectable with deep-sequencing methods. We have sequenced expressed mRNAs from actively photosynthesizing, starved individuals of two photosynthetic sea slug species, Plakobranchus ocellatus Van Hasselt, 1824 and Elysia timida Risso, 1818. We find that nuclear-encoded, algal-derived genes specific to photosynthetic function are expressed neither in P. ocellatus nor in E. timida. Despite their dramatic plastid longevity, these photosynthetic sacoglossan slugs do not express genes acquired from algal nuclei in order to maintain plastid function.

From sea to land and beyond – New insights into the evolution of euthyneuran Gastropoda (Mollusca)

BackgroundThe Euthyneura are considered to be the most successful and diverse group of Gastropoda. Phylogenetically, they are riven with controversy. Previous morphology-based phylogenetic studies have been greatly hampered by rampant parallelism in morphological characters or by incomplete taxon sampling. Based on sequences of nuclear 18S rRNA and 28S rRNA as well as mitochondrial 16S rRNA and COI DNA from 56 taxa, we reconstructed the phylogeny of Euthyneura utilising Maximum Likelihood and Bayesian inference methods. The evolution of colonization of freshwater and terrestrial habitats by pulmonate Euthyneura, considered crucial in the evolution of this group of Gastropoda, is reconstructed with Bayesian approaches.ResultsWe found several well supported clades within Euthyneura, however, we could not confirm the traditional classification, since Pulmonata are paraphyletic and Opistobranchia are either polyphyletic or paraphyletic with several clades clearly distinguishable. Sacoglossa appear separately from the rest of the Opisthobranchia as sister taxon to basal Pulmonata. Within Pulmonata, Basommatophora are paraphyletic and Hygrophila and Eupulmonata form monophyletic clades. Pyramidelloidea are placed within Euthyneura rendering the Euthyneura paraphyletic.ConclusionBased on the current phylogeny, it can be proposed for the first time that invasion of freshwater by Pulmonata is a unique evolutionary event and has taken place directly from the marine environment via an aquatic pathway. The origin of colonisation of terrestrial habitats is seeded in marginal zones and has probably occurred via estuaries or semi-terrestrial habitats such as mangroves.

Flashback and foreshadowing—a review of the taxon Opisthobranchia

Opisthobranchia have experienced an unsettled taxonomic history. At the moment their taxonomy is in state of dramatic flux as recent phylogenetic studies have revealed traditional Opisthobranchia to be paraphyletic or even polyphyletic, allocating some traditional opisthobranch taxa to other groups of Heterobranchia, e.g. Pulmonata. Here we review the history of Opisthobranchia and their subgroups, explain their traditionally proposed relationships, and outline the most recent phylogenetic analyses based on various methods (morphology, single gene and multiple gene analyses, as well as genomic data). We also present a phylogenetic hypothesis on Heterobranchia that, according to the latest results, represents a consensus and is the most probable one available to date. The proposed phylogeny supports the Acteonoidea outside of monophyletic Euthyneura, the basal euthyneuran split into Nudipleura (Nudibranchia plus Pleurobranchoidea) and the recently established taxon Tectipleura. The latter divides into the Euopisthobranchia, containing most of the major traditional opisthobranch clades, and the Panpulmonata, with a mix of the former opisthobranch, putative allogastropod and pulmonate taxa. This “new euthyneuran tree” rejects the traditional taxa Opisthobranchia and Pulmonata, and, in particular, has profound implications for preconceived textbook scenarios of opisthobranch and pulmonate evolution, which must now be reconsidered. In the absence of systematic barriers, research communities—which have traditionally investigated marine and non-marine heterobranchs separately—need to interact and finally merge for the sake of science.

The taxonomist - an endangered race. A practical proposal for its survival.

BackgroundTaxonomy or biological systematics is the basic scientific discipline of biology, postulating hypotheses of identity and relationships, on which all other natural sciences dealing with organisms relies. However, the scientific contributions of taxonomists have been largely neglected when using species names in scientific publications by not citing the authority on which they are based.DiscussionConsequences of this neglect is reduced recognition of the importance of taxonomy, which in turn results in diminished funding, lower interest from journals in publishing taxonomic research, and a reduced number of young scientists entering the field. This has lead to the so-called taxonomic impediment at a time when biodiversity studies are of critical importance.Here we emphasize a practical and obvious solution to this dilemma. We propose that whenever a species name is used, the author(s) of the species hypothesis be included and the original literature source cited, including taxonomic revisions and identification literature - nothing more than what is done for every other hypothesis or assumption included in a scientific publication. In addition, we postulate that journals primarily publishing taxonomic studies should be indexed in ISISM.SummaryThe proposal outlined above would make visible the true contribution of taxonomists within the scientific community, and would provide a more accurate assessment for funding agencies impact and importance of taxonomy, and help in the recruitment of young scientists into the field, thus helping to alleviate the taxonomic impediment. In addition, it would also make much of the biological literature more robust by reducing or alleviating taxonomic uncertainty.

A new approach to an old conundrum—DNA barcoding sheds new light on phenotypic plasticity and morphological stasis in microsnails (Gastropoda, Pulmonata, Carychiidae)

The identification of microsnail taxa based on morphological characters is often a time‐consuming and inconclusive process. Aspects such as morphological stasis and phenotypic plasticity further complicate their taxonomic designation. In this study, we demonstrate that the application of DNA barcoding can alleviate these problems within the Carychiidae (Gastropoda, Pulmonata). These microsnails are a taxon of the pulmonate lineage and most likely migrated onto land independently of the Stylommatophora clade. Their taxonomical classification is currently based on conchological and anatomical characters only. Despite much confusion about historic species assignments, the Carychiidae can be unambiguously subdivided into two taxa: (i) Zospeum species, which are restricted to karst caves, and (ii) Carychium species, which occur in a broad range of environmental conditions. The implementation of discrete molecular data (COI marker) enabled us to correctly designate 90% of the carychiid microsnails. The remaining cases were probably cryptic Zospeum and Carychium taxa and incipient species, which require further investigation into their species status. Because conventional reliance upon mostly continuous (i.e. nondiscrete) conchological characters is subject to fallibility for many gastropod species assignments, we highly recommend the use of DNA barcoding as a taxonomic, cutting‐edge method for delimiting microsnail taxa.

Out of Antarctica? – New insights into the phylogeny and biogeography of the Pleurobranchomorpha (Mollusca, Gastropoda)

The aim of the current study was to gain new insights into the phylogeny, biogeography and evolution of the opisthobranch clade Pleurobranchomorpha. We focused on testing the hypothesis of an Antarctic origin of this clade. The combination of four gene markers (18S rDNA, 28S rDNA, 16S rDNA and CO1) was used to infer a phylogenetic hypothesis of the Pleurobranchomorpha employing Maximum likelihood and Bayesian inference methods. Four methodologically distinct approaches were applied to reconstruct the historical biogeography and dating of the tree was performed via relaxed molecular clock analysis. Phylogenetic analyses supported the monophyly of the Pleurobranchomorpha and their sister group relationship to the Nudibranchia. Monophyly of the main subgroups Pleurobranchaeinae and Pleurobranchinae could not be revealed. Reconstruction of the ancestral area of the Pleurobranchomorpha yielded different possibilities in the diverse analyses. However, the Pleurobranchinae most probably derived from an Antarctic origin. Estimation of divergence times revealed a long credible interval for the Pleurobranchomorpha, whereas the Pleurobranchinae diverged in Early Oligocene and underwent rapid radiation during Oligocene and Early Miocene. Divergence of the Pleurobranchinae into the Antarctic Tomthompsonia and the remaining species in Early Oligocene coincides with two major geological events; namely the onset of glaciation in Antarctica and the opening of the Drake Passage with following formation of the Antarctic Circumpolar Current (ACC). These sudden and dramatic changes in climate probably led to subsequent migration of the last common ancestor of the remaining Pleurobranchinae into warmer regions, while the ACC may have accounted for larval dispersal to the Eastern Atlantic.

Evolution of microgastropods (Ellobioidea, Carychiidae): integrating taxonomic, phylogenetic and evolutionary hypotheses.

BackgroundCurrent biodiversity patterns are considered largely the result of past climatic and tectonic changes. In an integrative approach, we combine taxonomic and phylogenetic hypotheses to analyze temporal and geographic diversification of epigean (Carychium) and subterranean (Zospeum) evolutionary lineages in Carychiidae (Eupulmonata, Ellobioidea). We explicitly test three hypotheses: 1) morphospecies encompass unrecognized evolutionary lineages, 2) limited dispersal results in a close genetic relationship of geographical proximally distributed taxa and 3) major climatic and tectonic events had an impact on lineage diversification within Carychiidae.ResultsInitial morphospecies assignments were investigated by different molecular delimitation approaches (threshold, ABGD, GMYC and SP). Despite a conservative delimitation strategy, carychiid morphospecies comprise a great number of unrecognized evolutionary lineages. We attribute this phenomenon to historic underestimation of morphological stasis and phenotypic variability amongst lineages. The first molecular phylogenetic hypothesis for the Carychiidae (based on COI, 16S and H3) reveals Carychium and Zospeum to be reciprocally monophyletic. Geographical proximally distributed lineages are often closely related. The temporal diversification of Carychiidae is best described by a constant rate model of diversification. The evolution of Carychiidae is characterized by relatively few (long distance) colonization events. We find support for an Asian origin of Carychium. Zospeum may have arrived in Europe before extant members of Carychium. Distantly related Carychium clades inhabit a wide spectrum of the available bioclimatic niche and demonstrate considerable niche overlap.ConclusionsCarychiid taxonomy is in dire need of revision. An inferred wide distribution and variable phenotype suggest underestimated diversity in Zospeum. Several Carychium morphospecies are results of past taxonomic lumping. By collecting populations at their type locality, molecular investigations are able to link historic morphospecies assignments to their respective evolutionary lineage. We propose that rare founder populations initially colonized a continent or cave system. Subsequent passive dispersal into adjacent areas led to in situ pan-continental or mountain range diversifications. Major environmental changes did not influence carychiid diversification. However, certain molecular delimitation methods indicated a recent decrease in diversification rate. We attribute this decrease to protracted speciation.

Comparative investigation of the genital systems in the Opisthobranchia (Mollusca, Gastropoda) with special emphasis on the nidamental glandular system

Abstract The reproductive systems and especially the nidamental glands of 20 species of Opisthobranchia belonging to the ”Cephalaspidea s. l.”, Anaspidea, Sacoglossa, Tylodinoidea and Pleurobranchoidea, have been investigated histologically and ultrastructurally. The nidamental glandular system is responsible for the formation of the egg masses. In all investigated species it is divided into three distinct parts. The most proximal part can be an albumen gland (some ”Cephalaspidea s. l.”, Anaspidea and Sacoglossa) or can exhibit a capsule gland (some ”Cephalaspidea s. l.”, Tylodinoidea and Pleurobranchoidea). All species additionally possess a membrane gland and a distally lying mucous gland. In some species the most distal part of the oviduct was also found to be glandular. The structure of the nidamental glands is described and compared within the Opisthobranchia. Albumen and capsule glands are found to be homologous glandular parts of the system. It can be concluded that the albumen gland has undergone a structural and functional change within the evolution of the Opisthobranchia.

On the Fine Structure of Opisthobranch Egg Masses (Mollusca, Gastropoda)

Abstract The light-microscopic structure and ultrastructure of the egg masses of some species of Opisthobranchia (Mollusca, Gastropoda) are presented and discussed. The egg masses have a rather uniform structure. The embryos are embedded in a viscous mass of albumen in some species of “Cephalaspidea s. l.”, all Anaspidea and Sacoglossa, whereas in other taxa examined (some “Cephalaspidea s. l.”, and Nudibranchia) this albumen is missing in the egg capsules. Instead, the albumen forms an additional compact albuminous layer. This layer seems to dissolve during intracapsular development and is apparently ingested by the veliger larvae in the capsules. Embryos and albumen, or embryos and albuminous layers are enveloped by a mucoid membrane, forming distinct capsules. The capsules themselves are surrounded by an inner mucous layer and embedded in a mucous matrix. Along the outside of the egg mass a multi-layered outer mucous cover is present. Interspecific differences in this general structure are apparent in the number of embryos per capsule, the thickness of the capsule, the fine structure of the capsule and the density and fine structure of the mucous layers.