<italic toggle="yes">In Vitro</italic> Culture of the Insect Endosymbiont <italic toggle="yes">Spiroplasma poulsonii</italic> Highlights Bacterial Genes Involved in Host-Symbiont Interaction

ABSTRACT Endosymbiotic bacteria associated with eukaryotic hosts are omnipresent in nature, particularly in insects. Studying the bacterial side of host-symbiont interactions is, however, often limited by the unculturability and genetic intractability of the symbionts. Spiroplasma poulsonii is a mat...

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Autores principales:	Florent Masson, Sandra Calderon Copete, Fanny Schüpfer, Gonzalo Garcia-Arraez, Bruno Lemaitre
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	Spiroplasma endosymbiosis host-symbiont interaction Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c21a3ee76c4c47f2b60ae1966e7ef511
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spelling	oai:doaj.org-article:c21a3ee76c4c47f2b60ae1966e7ef5112021-11-15T15:53:26Z<italic toggle="yes">In Vitro</italic> Culture of the Insect Endosymbiont <italic toggle="yes">Spiroplasma poulsonii</italic> Highlights Bacterial Genes Involved in Host-Symbiont Interaction10.1128/mBio.00024-182150-7511https://doaj.org/article/c21a3ee76c4c47f2b60ae1966e7ef5112018-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00024-18https://doaj.org/toc/2150-7511ABSTRACT Endosymbiotic bacteria associated with eukaryotic hosts are omnipresent in nature, particularly in insects. Studying the bacterial side of host-symbiont interactions is, however, often limited by the unculturability and genetic intractability of the symbionts. Spiroplasma poulsonii is a maternally transmitted bacterial endosymbiont that is naturally associated with several Drosophila species. S. poulsonii strongly affects its host’s physiology, for example by causing male killing or by protecting it against various parasites. Despite intense work on this model since the 1950s, attempts to cultivate endosymbiotic Spiroplasma in vitro have failed so far. Here, we developed a method to sustain the in vitro culture of S. poulsonii by optimizing a commercially accessible medium. We also provide a complete genome assembly, including the first sequence of a natural plasmid of an endosymbiotic Spiroplasma species. Last, by comparing the transcriptome of the in vitro culture to the transcriptome of bacteria extracted from the host, we identified genes putatively involved in host-symbiont interactions. This work provides new opportunities to study the physiology of endosymbiotic Spiroplasma and paves the way to dissect insect-endosymbiont interactions with two genetically tractable partners. IMPORTANCE The discovery of insect bacterial endosymbionts (maternally transmitted bacteria) has revolutionized the study of insects, suggesting novel strategies for their control. Most endosymbionts are strongly dependent on their host to survive, making them uncultivable in artificial systems and genetically intractable. Spiroplasma poulsonii is an endosymbiont of Drosophila that affects host metabolism, reproduction, and defense against parasites. By providing the first reliable culture medium that allows a long-lasting in vitro culture of Spiroplasma and by elucidating its complete genome, this work lays the foundation for the development of genetic engineering tools to dissect endosymbiosis with two partners amenable to molecular study. Furthermore, the optimization method that we describe can be used on other yet uncultivable symbionts, opening new technical opportunities in the field of host-microbes interactions.Florent MassonSandra Calderon CopeteFanny SchüpferGonzalo Garcia-ArraezBruno LemaitreAmerican Society for MicrobiologyarticleSpiroplasmaendosymbiosishost-symbiont interactionMicrobiologyQR1-502ENmBio, Vol 9, Iss 2 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Spiroplasma endosymbiosis host-symbiont interaction Microbiology QR1-502
spellingShingle	Spiroplasma endosymbiosis host-symbiont interaction Microbiology QR1-502 Florent Masson Sandra Calderon Copete Fanny Schüpfer Gonzalo Garcia-Arraez Bruno Lemaitre <italic toggle="yes">In Vitro</italic> Culture of the Insect Endosymbiont <italic toggle="yes">Spiroplasma poulsonii</italic> Highlights Bacterial Genes Involved in Host-Symbiont Interaction
description	ABSTRACT Endosymbiotic bacteria associated with eukaryotic hosts are omnipresent in nature, particularly in insects. Studying the bacterial side of host-symbiont interactions is, however, often limited by the unculturability and genetic intractability of the symbionts. Spiroplasma poulsonii is a maternally transmitted bacterial endosymbiont that is naturally associated with several Drosophila species. S. poulsonii strongly affects its host’s physiology, for example by causing male killing or by protecting it against various parasites. Despite intense work on this model since the 1950s, attempts to cultivate endosymbiotic Spiroplasma in vitro have failed so far. Here, we developed a method to sustain the in vitro culture of S. poulsonii by optimizing a commercially accessible medium. We also provide a complete genome assembly, including the first sequence of a natural plasmid of an endosymbiotic Spiroplasma species. Last, by comparing the transcriptome of the in vitro culture to the transcriptome of bacteria extracted from the host, we identified genes putatively involved in host-symbiont interactions. This work provides new opportunities to study the physiology of endosymbiotic Spiroplasma and paves the way to dissect insect-endosymbiont interactions with two genetically tractable partners. IMPORTANCE The discovery of insect bacterial endosymbionts (maternally transmitted bacteria) has revolutionized the study of insects, suggesting novel strategies for their control. Most endosymbionts are strongly dependent on their host to survive, making them uncultivable in artificial systems and genetically intractable. Spiroplasma poulsonii is an endosymbiont of Drosophila that affects host metabolism, reproduction, and defense against parasites. By providing the first reliable culture medium that allows a long-lasting in vitro culture of Spiroplasma and by elucidating its complete genome, this work lays the foundation for the development of genetic engineering tools to dissect endosymbiosis with two partners amenable to molecular study. Furthermore, the optimization method that we describe can be used on other yet uncultivable symbionts, opening new technical opportunities in the field of host-microbes interactions.
format	article
author	Florent Masson Sandra Calderon Copete Fanny Schüpfer Gonzalo Garcia-Arraez Bruno Lemaitre
author_facet	Florent Masson Sandra Calderon Copete Fanny Schüpfer Gonzalo Garcia-Arraez Bruno Lemaitre
author_sort	Florent Masson
title	<italic toggle="yes">In Vitro</italic> Culture of the Insect Endosymbiont <italic toggle="yes">Spiroplasma poulsonii</italic> Highlights Bacterial Genes Involved in Host-Symbiont Interaction
title_short	<italic toggle="yes">In Vitro</italic> Culture of the Insect Endosymbiont <italic toggle="yes">Spiroplasma poulsonii</italic> Highlights Bacterial Genes Involved in Host-Symbiont Interaction
title_full	<italic toggle="yes">In Vitro</italic> Culture of the Insect Endosymbiont <italic toggle="yes">Spiroplasma poulsonii</italic> Highlights Bacterial Genes Involved in Host-Symbiont Interaction
title_fullStr	<italic toggle="yes">In Vitro</italic> Culture of the Insect Endosymbiont <italic toggle="yes">Spiroplasma poulsonii</italic> Highlights Bacterial Genes Involved in Host-Symbiont Interaction
title_full_unstemmed	<italic toggle="yes">In Vitro</italic> Culture of the Insect Endosymbiont <italic toggle="yes">Spiroplasma poulsonii</italic> Highlights Bacterial Genes Involved in Host-Symbiont Interaction
title_sort	<italic toggle="yes">in vitro</italic> culture of the insect endosymbiont <italic toggle="yes">spiroplasma poulsonii</italic> highlights bacterial genes involved in host-symbiont interaction
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/c21a3ee76c4c47f2b60ae1966e7ef511
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<italic toggle="yes">In Vitro</italic> Culture of the Insect Endosymbiont <italic toggle="yes">Spiroplasma poulsonii</italic> Highlights Bacterial Genes Involved in Host-Symbiont Interaction

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