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Reverse taxonomy: an approach towards determining the diversity of meiobenthic organisms based on ribosomal RNA signature sequences.

ABSTRACT: Organisms living in or on the sediment layer of water bodies constitute the benthos fauna, which is known to harbour a large number of species of diverse taxonomic groups. The benthos plays a significant role in the nutrient cycle and it is, therefore, of high ecological relevance. Here, we have explored a DNA-taxonomic approach to access the meiobenthic organismic diversity, by focusing on obtaining signature sequences from a part of the large ribosomal subunit rRNA (28S), the D3-D5 region. To obtain a broad representation of taxa, benthos samples were taken from 12 lakes in Germany, representing different ecological conditions. In a first approach, we have extracted whole DNA from these samples, amplified the respective fragment by PCR, cloned the fragments and sequenced individual clones. However, we found a relatively large number of recombinant clones that must be considered PCR artefacts. In a second approach we have, therefore, directly sequenced PCR fragments that were obtained from DNA extracts of randomly picked individual organisms. In total, we have obtained 264 new unique sequences, which can be readily placed into taxon groups, based on phylogenetic comparison with currently available database sequences. The group with the highest taxon abundance were nematodes and protozoa, followed by chironomids. However, we find also that we have by far not exhausted the diversity of organisms in the samples. Still, our data provide a framework within which a meiobenthos DNA signature sequence database can be constructed, that will allow to develop the necessary techniques for studying taxon diversity in the context of ecological analysis. Since many taxa in our analysis are initially only identified via their signature sequences, but not yet their morphology, we propose to call this approach 'reverse taxonomy'.

SUBMITTER: Markmann M

PROVIDER: S-EPMC1609229 | biostudies-literature | 2005 Oct

REPOSITORIES: biostudies-literature

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Reverse taxonomy: an approach towards determining the diversity of meiobenthic organisms based on ribosomal RNA signature sequences.

Markmann Melanie M Tautz Diethard D

Philosophical transactions of the Royal Society of London. Series B, Biological sciences 20051001 1462

Organisms living in or on the sediment layer of water bodies constitute the benthos fauna, which is known to harbour a large number of species of diverse taxonomic groups. The benthos plays a significant role in the nutrient cycle and it is, therefore, of high ecological relevance. Here, we have explored a DNA-taxonomic approach to access the meiobenthic organismic diversity, by focusing on obtaining signature sequences from a part of the large ribosomal subunit rRNA (28S), the D3-D5 region. To ...[more]

PMID: 16214749

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Project description:BackgroundThe molecular operational taxonomic unit (MOTU) has recently been applied to microbial and microscopic animal biodiversity surveys. However, in many cases, some of the MOTUs cannot be definitively tied to any of the taxonomic groups in current databases. To surmount these limitations, the concept of "reverse taxonomy" has been proposed, i.e. to primarily list the MOTUs with morphological information, and then identify and/or describe them at genus/species level using subsamples or by re-isolating the target organisms. Nevertheless, the application of "reverse taxonomy" has not been sufficiently evaluated. Therefore, the practical applicability of "reverse taxonomy" is tested using termite-associated nematodes as a model system for phoretic/parasitic organisms which have high habitat specificity and a potential handle (their termite host species) for re-isolation attempts.MethodologyForty-eight species (from 298 colonies) of termites collected from the American tropics and subtropics were examined for their nematode associates using the reverse taxonomy method and culturing attempts (morphological identification and further sequencing efforts). The survey yielded 51 sequence types (?=? MOTUs) belonging to 19 tentatively identified genera. Within these, four were identified based on molecular data with preliminary morphological observation, and an additional seven were identified or characterized from successful culturing, leaving eight genera unidentified.ConclusionsThat 1/3 of the genera were not successfully identified suggests deficiencies in the depth of available sequences in the database and biological characters, i.e. usually isolated as phoretic/parasitic stages which are not available for morphological identification, and too many undiscovered lineages of nematodes. Although there still is the issue of culturability of nematodes, culturing attempts could help to make reverse taxonomy methods more effective. However, expansion of the database, i.e., production of more DNA barcodes tied to biological information by finding and characterizing additional new and known lineages, is necessary for analyzing functional diversity.

Project description:AimWe evaluated traditional biogeographic boundaries of coastal marine regions in Southwestern Atlantic using DNA sequence data from common, rocky-shore inhabiting, marine mites of the genera Agauopsis and Rhombognathus, family Halacaridae.MethodsWe investigated geographic population genetic structure using CO1 gene sequences, estimated divergence times using a multigene dataset and absolute time-calibrated molecular clock analyses, and performed environmental niche modeling (ENM) of common marine mite species.ResultsAgauopsis legionium has a shallow history (2.01 Ma) with four geographically differentiated groups. Two of them corresponded to the traditional Amazonian and Northeastern ecoregions, but the boundary between the two other groups was inferred at the Abrolhos Plateau, not Cabo Frio. Rhombognathus levigatoides s. lat. was represented by two cryptic species that diverged 7.22 (multilocus data) or 10.01 Ma (CO1-only analyses), with their boundary, again at the Abrolhos Plateau. ENM showed that A. legionium has suitable habitats scattered along the coast, while the two R. levigatoides cryptic species differ considerably in their niches, especially in parameters related to upwelling. This indicates that genetic isolation associated with the Abrolhos Plateau occurred in both lineages, but for the R. levigatoides species complex, ecological niche specialization was also an important factor.Main conclusionsOur study suggests that the major biogeographic boundary in the Southwestern Atlantic lies not at Cabo Frio but at the Abrolhos Plateau. There two biogeographically relevant factors meet (a) changes in current directions (which limit dispersal) and (b) abrupt changes in environmental parameters associated with the South Atlantic Central Waters (SACW) upwelling (offering distinct ecological niches). We suggest that our result represents a general biogeographic pattern because a barrier at the Abrolhos Plateau was found previously for the fish genus Macrodon (phylogeographic data), prosobranch mollusks, ascidians, and reef fishes (community-level data).

Dataset Information

Reverse taxonomy: an approach towards determining the diversity of meiobenthic organisms based on ribosomal RNA signature sequences.

Publications

Reverse taxonomy: an approach towards determining the diversity of meiobenthic organisms based on ribosomal RNA signature sequences.

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