KinoRoot was divided into three main work packages:
- Study the morphology of weakly segmented kinorhynchs
We examined traces of segmentation in three potentially unsegmented kinorhynch genera through immunohistochemical, histological, and ultrastructural examinations of muscular and nervous systems combined with computational 3D reconstruction.
Previously collected, identified and fixed target species of the genera Cateria, Franciscideres, and Zelinkaderes were stained using specific markers for muscles (phalloidin) and to characterize different subsets of the nervous system we used antibodies against alpha-tubulin, serotonin, and FMRFamide combined with nuclear labeling (DAPI). Tracing and labeling muscle sets and three-dimensional models based on the CLSM scans were built for each of the species. Immunohistochemical analyses were complemented with additional histological, scanning electron microscopy, or transmission electron microscopy studies.
- Analysis of phylogenetic relationships in Kinorhyncha.
We collected 18 species from 15 key genera representing half of the diversity of the group, extracted RNA from single specimens, sequenced, de novo assembled their transcriptomes. Generate phylogenomic trees to explain the evolutionary relationships within the phylum.
- Reconstruct the evolution of segmentation.
Based on combined results from the morphological and molecular analyses the evolution of kinorhynch segmentation was reconstructed on the obtained phylogenomic tree.
Results and exploitation:
- Comprehensive morphological database (including 3D models) of the nervous system and musculature of five kinorhynch genera, three of them “less segmented”. These will be used as a reference for any future comparative morphological study within the phylum and outside by any researcher interested in the evolution of organ systems.
- Establishment and optimization of a protocol to successfully extract RNA from single individuals
- Generation of 18 new high-quality kinorhynch transcriptomes representing 15 genera inclusive 3 of the weakly segmented genera. This will be useful for testing hypotheses within Kinorhyncha and for future, large-scaled phylogenomic or comparative transcriptomic analyses.
- Production for the first time of a robust phylogenomic tree resolving most of the internal relationships in kinorhynchs inclusive the position of aberrant, less segmented genera.
- Resolved evolution of segmentation within kinorhynchs and proposal of a scenario for the evolution of segmentation in Ecdysozoa. Understanding the evolution of this body plan is pivotal for understanding the evolution of most other organ systems. Therefore, KinoRoot results are relevant and useful for a broad range of evolutionary scientists, paleontologists, and invertebrate zoologists.
Dissemination:
To date I have published 2 research papers during the development of this action, one more is under review and I am currently in the process of publishing a fourth one. I attended one international conference during the action, another one just before the start of the action introducing my project, and I intend to attend an additional one (which was postponed) after the action to present the final results. Additional dissemination of my results was through participation as a speaker in several seminars at the Natural History Museum of Demark and The BIO department at the University of Copenhagen.
