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Microbiology 154 (2008), 1454-1463; DOI  10.1099/mic.0.2007/013888-0
© 2008 Society for General Microbiology

A chromosome map of the Flavescence dorée phytoplasma

Sylvie Malembic-Maher1,2,{dagger}, Fiona Constable3,4,5,{dagger},{ddagger}, Agnès Cimerman1,2, Guillaume Arnaud1,2, Patricia Carle1,2, Xavier Foissac1,2 and Elisabeth Boudon-Padieu3,4,5

1 INRA, UMR1090 Génomique Diversité Pouvoir Pathogène, F-33883 Villenave d'Ornon, France
2 Université Bordeaux 2, UMR1090 Génomique Diversité Pouvoir Pathogène, F-33076 Bordeaux, France
3 INRA, UMR1088 Plante Microbe Environnement, F-21065 Dijon, France
4 Université de Bourgogne, UMR1088 Plante Microbe Environnement, F-21000 Dijon, France
5 CNRS, UMR1088 Plante Microbe Environnement, F-21000 Dijon, France

Correspondence
Sylvie Malembic-Maher
smalembi{at}bordeaux.inra.fr

The Flavescence dorée phytoplasma (FD-P), a non-cultivable, plant-pathogenic bacterium of the class Mollicutes, is the causal agent of a quarantine disease affecting vineyards of southern Europe, mainly in southern France and northern Italy. To investigate FD-P diversity and phytoplasma genetic determinants governing the FD-P life cycle, a genome project has been initiated. A physical map of the chromosome of FD-P strain FD92, purified from infected broad beans, was constructed by performing restriction digests of the chromosome and resolving the fragments by PFGE. Single and double digestions of the chromosome with the enzymes SalI, BssHII, MluI and EagI were performed and used to map 13 restriction sites on the FD-P chromosome. The size of the chromosome was calculated to be 671 kbp. Southern blot analyses using cloned phytoplasma probes were carried out to assist in the arrangement of contiguous restriction fragments and to map eight genetic loci, including the two rRNA operons, the tuf, uvrB-degV and secY-map (FD9) genes, the FD2 marker and two orphan sequences (FDDH29 and FDSH05) isolated through subtractive suppression hybridization.

Abbreviations: FD, Flavescence dorée; FD-P, Flavescence dorée phytoplasma; SSH, subtractive suppression hybridization

{dagger}These authors contributed equally to this paper.

{ddagger}Present address: Department of Primary Industries, Knoxfield, Private Bag 15, Ferntree Gully Delivery Centre, Victoria 3156, Australia.






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