Coelomocyte‐derived fluorescence and DNA markers of composting earthworm species. Issue 1 (30th September 2013)

• Record Type: Journal Article
• Authors: Rorat, Agnieszka; Kachamakova‐Trojanowska, Neli; Jozkowicz, Alicja; Kruk, Jerzy; Cocquerelle, Claude; Vandenbulcke, Franck; Santocki, Michal; Plytycz, Barbara
• Is Part Of: Journal of experimental zoology. Volume 321:Issue 1(2014)
• Page Start: 28
• Page End: 40
• Publication Date: 2013-09-30
• Subjects: Zoology -- Periodicals; Ecological genetics -- Periodicals; Ecophysiology -- Periodicals; 571.105
• DOI: 10.1002/jez.1834
• Languages: English
• ISSNs: 1932-5223

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>ABSTRACT</title> <sec id="jez1834-sec-0001" sec-type="section"> <p>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 <italic>Eisenia andrei</italic>, <italic>Eisenia fetida</italic>, and <italic>Dendrobaena veneta</italic>, and left for reproduction and analysis of non‐invasively retrieved coelomocyte‐containing coelomic fluid and/or species‐specific partial sequences of cytochrome <italic>c</italic> 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 <italic>D. veneta</italic> ≪ <italic>E. andrei</italic> < <italic>E. fetida</italic>, and intensity of eleocyte‐derived fluorescence is lower in <italic>D. veneta</italic> than in <italic>Eisenia</italic> spp. Spectrofluorimetry of coelomocyte lysates revealed that the amount of eleocyte‐stored riboflavin is significantly lower in coelomocyte lysates from <italic>D. veneta</italic> than from <italic>Eisenia</italic> spp., and the emission peak of X‐fluorophore is much more distinct in <italic>D. veneta</italic> than in <italic>Eisenia</italic> spp. Coelomic fluid of <italic>E. andrei</italic> exhibits a very distinct spectra of MUG fluorophore which are absent in <italic>D. veneta</italic> and in the majority of <italic>E. fetida</italic>, while some <italic>E. fetida</italic> 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.</p> </sec> </abstract>