Conflicts of interest: None.
Coelomocyte-derived fluorescence and DNA markers of composting earthworm species
Article first published online: 30 SEP 2013
© 2013 Wiley Periodicals, Inc.
Journal of Experimental Zoology Part A: Ecological Genetics and Physiology
Volume 321, Issue 1, pages 28–40, January 2014
How to Cite
2014. Coelomocyte-derived fluorescence and DNA markers of composting earthworm species. J. Exp. Zool. 321A:28–40., , , , , , , .
- Issue published online: 9 DEC 2013
- Article first published online: 30 SEP 2013
- Manuscript Accepted: 4 SEP 2013
- Manuscript Revised: 16 JUL 2013
- Manuscript Received: 3 APR 2013
- Ministry of Science and Higher Education of Poland. Grant Number: K/ZDS/001955
- French Agence National de la Recherche (ANR)
- CESA Interconnect project
Supravital species identification of morphologically similar syntopic earthworms inhabiting dung and compost heaps or those from commercial cultures is difficult. The aim of the studies was to find out non-invasive species-specific markers for proper segregation of earthworm species from a dense mixed colony of waste decomposers. Worms were segregated according to external characteristics into Eisenia andrei, Eisenia fetida, and Dendrobaena veneta, and left for reproduction and analysis of non-invasively retrieved coelomocyte-containing coelomic fluid and/or species-specific partial sequences of cytochrome c oxidase subunit I (COI) gene in DNA extracted from amputated tail tips of adults and their offspring. Flow cytometric analysis of coelomocyte samples revealed that amount of nuclear DNA increases in order D. veneta ≪ E. andrei < E. fetida, and intensity of eleocyte-derived fluorescence is lower in D. veneta than in Eisenia spp. Spectrofluorimetry of coelomocyte lysates revealed that the amount of eleocyte-stored riboflavin is significantly lower in coelomocyte lysates from D. veneta than from Eisenia spp., and the emission peak of X-fluorophore is much more distinct in D. veneta than in Eisenia spp. Coelomic fluid of E. andrei exhibits a very distinct spectra of MUG fluorophore which are absent in D. veneta and in the majority of E. fetida, while some E. fetida possess MUG-like fluorophore. Sequences of the COI gene in the DNA of the worms from the mixed colony and their offspring confirmed species identity. In conclusion, species-specific coelomocyte-derived markers may be a useful complement to morphological and DNA-based taxonomy during studies on syntopic earthworms. J. Exp. Zool. 321A: 28–40, 2014. © 2013 Wiley Periodicals, Inc.