Evaluation of Enrichment Protocols for Bacterial Endosymbionts of Ciliates by Real-Time PCR

Current Microbiology
Michele CastelliGiulio Petroni

Abstract

Large-scale studies on obligate bacterial endosymbionts may frequently require preliminary purification and enrichment protocols, which are often elaborate to set up and to evaluate, especially if the host organism is a protist. The purpose of this study was to develop a real-time PCR-based strategy and employ it for assessing two of such enrichment protocols for Holospora caryophila, hosted by the ciliate Paramecium. Four SSU rRNA gene-targeted real-time PCR assays were designed, which allowed to compare the amount of H. caryophila to other organisms, namely the host, its food bacterium (Raoultella planticola), and free-living bacteria present in the culture medium. By the use of the real-time PCR assays in combination, it was possible to conclude that the "cell fractionation" protocol was quite successful in the enrichment of the symbiont, while the "Percoll gradient" protocol will need further refinements to be fully repeatable. The proposed approach has the potential to facilitate and encourage future studies on the yet underexplored field of bacterial endosymbionts of ciliates and other protists. It can also find valuable applications for experimental questions other than those tested, such as fast and precise assessment o...Continue Reading

References

Jul 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·R C Findly, J G Gall
Jul 21, 2004·International Review of Cytology·Sergei I Fokin
Jul 29, 2004·International Journal of Systematic and Evolutionary Microbiology·Claudia VanniniGiulio Petroni
Dec 7, 2005·FEMS Microbiology Ecology·Fei ZhuDaniel Vaulot
Aug 12, 2006·Cell·Colin Dale, Nancy A Moran
Aug 15, 2006·Molecular Plant-microbe Interactions : MPMI·Mónica Rosenblueth, Esperanza Martínez-Romero
May 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·Guido FaviaDaniele Daffonchio
Feb 13, 2008·Nature Reviews. Genetics·Andrés MoyaAmparo Latorre
Nov 14, 2008·Nucleic Acids Research·J R ColeJ M Tiedje
Feb 24, 2009·Protist·Elena V SabaneyevaIlya N Skovorodkin
Feb 4, 2010·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Eva C M Nowack, Michael Melkonian
Mar 30, 2011·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Julia Ferrari, Fabrice Vavre
Jan 10, 2012·Ecology Letters·Simon FellousOliver Kaltz
Mar 20, 2012·European Journal of Protistology·Sergei I Fokin
Apr 19, 2012·BMC Microbiology·Cindy M LiuLance B Price
Nov 30, 2012·Nucleic Acids Research·Christian QuastFrank Oliver Glöckner
Feb 13, 2013·The Quarterly Review of Biology·Scott F GilbertAlfred I Tauber
Aug 13, 2013·American Journal of Botany·Jonathan R GaieroKari E Dunfield
Aug 24, 2013·Proceedings. Biological Sciences·Alison B DuncanOliver Kaltz
Oct 30, 2013·Proceedings of the National Academy of Sciences of the United States of America·Vittorio BoscaroGiulio Petroni
Feb 5, 2014·Cold Spring Harbor Perspectives in Biology·Angela E Douglas
Sep 10, 2014·FEMS Microbiology Ecology·Alejandro BelancheCharles J Newbold
Oct 12, 2014·Journal of Molecular Biology·Matthew R Redinbo
Oct 13, 2014·Current Opinion in Microbiology·Verena ZimorskiSven B Gould
Jan 31, 2015·European Journal of Protistology·Michele CastelliGiulio Petroni

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