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Polyphasic study of Zymomonas mobilis strains revealing the existence of a novel subspecies Z. mobilis subsp. francensis subsp. nov., isolated from French cider

¹ ADRIA NORMANDIE, bvd du 13 juin 1944, 14310 Villers-Bocage, France
² USC-INRA Laboratoire de Microbiologie de l'Environnement, Université de Caen, Esplanade de la paix, 14000 Caen, France

Correspondence
Monika Coton
mcoton{at}adrianie.org

Zymomonas mobilis strains recently isolated from French ‘framboisé’ ciders were compared with collection strains of the two defined subspecies, Z. mobilis subsp. mobilis and Z. mobilis subsp. pomaceae, using a polyphasic approach. Six strains isolated from six different regions of France were compared with three strains of Z. mobilis subsp. mobilis, including the type strain LMG 404^T, and four strains of Z. mobilis subsp. pomaceae, including the type strain LMG 448^T, using phenotypic and genotypic methods. For phenotypic characterization, both physiological tests and SDS-PAGE protein profiles revealed significant differences between the two known subspecies and the French isolates; three distinct groups were observed. These findings were further confirmed by random amplified polymorphic DNA and repetitive extragenic palindromic-PCR genotyping methods in which the French isolates were clearly distinguished from the other two subspecies. Sequence analysis of a fragment ranging from 604 to 617 nucleotides corresponding to the 16S–23S rRNA gene intergenic spacer region (ISR), a 592 nucleotide HSP60 gene fragment and a 1044 nucleotide gyrB gene fragment confirmed the presence of three distinct groups. The French strains exhibited almost 94 % similarity to the ISR, 90 % to HSP60 and 86 % to gyrB sequences of the three collection strains of Z. mobilis subsp. mobilis and 87, 84 and 80 % sequence similarity, respectively, was observed with the four Z. mobilis subsp. pomaceae strains. Based on both the phenotypic and genotypic results, the French strains are proposed to represent a novel subspecies, Zymomonas mobilis subsp. francensis subsp. nov. Strain AN0101^T (=LMG 22974^T=CIP 108684^T) was designated as the type strain.

Published online ahead of print on 9 September 2005 as DOI 10.1099/ijs.0.63732-0.

The GenBank/EMBL/DDBJ accession numbers for the 16S–23S ISR sequences of Zymomonas mobilis strains LMG 404^T, LMG 445, AN0101^T, AN0102 and LMG 448^T are AY940084–AY940087, respectively. Those of the partial HSP60 gene sequences of strains LMG 404^T, LMG 445, LMG 448^T, LMG 5993 and AN0101^T are AY940079–AY940083, respectively, and those of the gyrB gene sequences for strains LMG 404^T, LMG 445, LMG 448^T and AN0101^T are AY940075–AY940078, respectively.

Figures showing the results of SDS-PAGE analysis and phylogenetic trees generated from the ISR, HSP60 and gyrB gene sequences are available as supplementary material in IJSEM Online.