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Differentiation of species of the family Acetobacteraceae by AFLP DNA fingerprinting: reclassification of Gluconacetobacter kombuchae as a later heterotypic synonym of Gluconacetobacter hansenii

¹ BCCM/LMG Bacteria Collection;
² Vrije Universiteit Brussel;
³ BCCM/LMG Bacteria Collection, Ghent University

⁴ E-mail: ilse.cleenwerck{at}ugent.be

Amplified fragment length polymorphism (AFLP) DNA fingerprinting was investigated as a tool for fast and accurate identification of acetic acid bacteria (AAB) to the species level. One hundred and thirty five reference strains and 15 additional strains, representing the 50 recognized species of the family Acetobacteraceae, were subjected to AFLP analysis using the restriction enzyme combination ApaI/TaqI and the primer combination A03/T03. The reference strains were previously subjected to either DNA-DNA hybridizations or 16S-23S rDNA spacer region sequence analysis and were regarded as being accurately classified to the species level. The present study revealed that six of these strains have to be reclassified, namely Gluconacetobacter europaeus LMG 1518 and Gluconacetobacter xylinus LMG 1510 as Gluconacetobacter xylinus and Gluconacetobacter europaeus, respectively; Gluconacetobacter kombuchae LMG 23726T as Gluconacetobacter hansenii; and Acetobacter orleanensis LMG 1545, LMG 1592 and LMG 1608 as Acetobacter cerevisiae. Cluster analysis of the AFLP DNA fingerprints of the reference strains revealed one cluster for each species, showing a linkage level below 50 % with other clusters, except for Acetobacter pasteurianus, Acetobacter indonesiensis, and Acetobacter cerevisiae. The latter species were separated into two, two, and three clusters, respectively. The AFLP data further supported to classify Gluconacetobacter oboediens and Gluconacetobacter intermedius, for which at present confusion exists concerning their taxonomic status, as different taxa. The 15 additional strains could all be identified to the species level. AFLP analysis further revealed that some species harbour genetically diverse strains, whereas other species consist of strains showing similar banding patterns, indicating a more limited genetic diversity. It can be concluded that AFLP DNA fingerprinting is suitable for accurate identification and classification of a broad range of AAB, as well as for determination of intraspecific genetic diversity.