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Journal of Bacteriology, October 2004, p. 6876-6884, Vol. 186, No. 20
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.20.6876-6884.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Isolation, Free-Living Capacities, and Genome Structure of "Candidatus Glomeribacter gigasporarum," the Endocellular Bacterium of the Mycorrhizal Fungus Gigaspora margarita

P. Jargeat,1,{dagger} C. Cosseau,2,{dagger} B. Ola'h,1 A. Jauneau,3 P. Bonfante,4 J. Batut,2 and G. Bécard1*

SCSV UMR 5546 CNRS-University Paul Sabatier,1 LIPM UMR 2594-441 CNRS-INRA,2 IFR 40, Castanet-Tolosan, France,3 Dipartimento di Biologia Vegetale dell'Università and Istituto per la Protezione delle Piante, CNR, Turin, Italy4

Received 8 April 2004/ Accepted 19 July 2004

"Candidatus Glomeribacter gigasporarum" is an endocellular ß-proteobacterium present in the arbuscular mycorrhizal (AM) fungus Gigaspora margarita. We established a protocol to isolate "Ca. Glomeribacter gigasporarum" from its host which allowed us to carry out morphological, physiological, and genomic investigations on purified bacteria. They are rod shaped, with a cell wall typical of gram-negative bacteria and a cytoplasm rich in ribosomes, and they present no flagella or pili. Isolated bacteria could not be grown in any of the 19 culture media tested, but they could be kept alive for up to 4 weeks. PCR-based investigations of purified DNA from isolated bacteria did not confirm the presence of all genes previously assigned to "Ca. Glomeribacter gigasporarum." In particular, the presence of nif genes could not be detected. Pulsed-field gel electrophoresis analyses allowed us to estimate the genome size of "Ca. Glomeribacter gigasporarum" to approximately 1.4 Mb with a ca. 750-kb chromosome and a 600- to 650-kb plasmid. This is the smallest genome known for a ß-proteobacterium. Such small genome sizes are typically found in endocellular bacteria living permanently in their host. Altogether, our data suggest that "Ca. Glomeribacter gigasporarum" is an ancient obligate endocellular bacterium of the AM fungus G. margarita.

* Corresponding author. Mailing address: UMR 5546 CNRS-University Paul Sabatier, 24, chemin de Borde-Rouge, BP 17, Auzeville 31326, Castanet-Tolosan, France. Phone: 33 562 19 35 20. Fax: 33 562 19 35 02. E-mail: becard{at}scsv.ups-tlse.fr.

{dagger} P.J. and C.C. contributed equally to the work.

Journal of Bacteriology, October 2004, p. 6876-6884, Vol. 186, No. 20
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.20.6876-6884.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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