Differentiation of two very similar glaucomid ciliate morphospecies (Ciliophora, Tetrahymenida) by fluorescence in situ hybridization with 18S rRNA targeted oligonucleotide probes.
Conventional, morphological identification of ciliates and other protozoa needs considerable experience and often is difficult as various staining methods must be applied. New molecular techniques, such as fluorescence in situ hybridization (FISH) with gene probes, are powerful means to overcome this problem. As a test case, the morphology of two very similar, and thus difficult to differentiate ciliate morphospecies, Glaucoma scintillans and Glaucomides bromelicola, were compared. They were then distinguished by applying the Ciliate-FISH technique with a set of eight 18S rRNA targeted oligonucleotide probes, four of which have been developed for specific detection of G. scintillans. The remaining four probes were designed to detect G. bromelicola in order to prove probe specificities by binding to the homologous target region of the probes mentioned before. The tests resulted in a clear and easy differentiation of the two species by fluorescence signals of three of the four tested probe pairs. Thus, FISH techniques are very useful for the identification and detection of protozoa and might be of great help studying geographical distributions of known taxa.
Technische Universität München, Lehrstuhl für Mikrobiologie, Am Hochanger 4, 85350 Freising, Germany. johannes.fried@uibk.ac.at
Johannes
J
Fried
Wilhelm
W
Foissner
D000818:Animals; D016798:Ciliophora / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000032:analysis* / Q000235:genetics
D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal; D012337:RNA, Ribosomal, 18S