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Genetic diversity within Lactobacillus sakei and Lactobacillus curvatus and design of PCR primers for its detection using randomly amplified polymorphic DNA

Francoise Berthier^1, and Stanislav D. Ehrlich²

¹ Laboratoire de Recherches sur la Viande and Laboratoire de Génétique Microbienne, Domaine de Vilvert, 78352 Jouy-en-Josas Cedex, France
² Laboratoire de Recherches sur la Viande, Institut National de la Recherche Agronomique, Domaine de Vilvert, 78352 Jouy-en-Josas Cedex, France

Author for correspondence: Francoise Berthier. Tel: +33 3 84 73 63 13. Fax: +33 3 84 37 37 81. e-mail: berthier{at}poligny.inra.fr

ABSTRACT

The genotypic and phenotypic diversity among isolates of the Lactobacillus curvatus/Lactobacillus graminis/Lactobacillus sakei group was evaluated by comparing RAPD data and results of biochemical tests, such as hydrolysis of arginine, D-lactate production, melibiose and xylose fermentation, and the presence of haem-dependent catalase. Analyses were applied to five type strains and to a collection of 165 isolates previously assigned to L. sakei or L. curvatus. Phenotypic and RAPD data were compared with each other and with previous DNA-DNA hybridization data. The phenotypic and genotypic separation between L. sakei, L. curvatus and L graminis was clear, and new insights into the detailed structure within L. sakei and L. curvatus were obtained. Individual strains could be typed by RAPD and, after the elimination of similar or identical isolates, two sub-groups in both L. curvatus and L. sakei were defined. The presence or absence of catalase activity further distinguished the two L. curvatus sub-groups. By cloning and sequencing specific RAPD products, pairs of PCR primers were developed that can be used to specifically detect L. curvatus, L. sakei and each of the L. sakei sub-groups.

Key Words: RAPD • PCR primers • genetic diversity • Lactobacillus sakei • Lactobacillus curvatus

Presant address: SRTAL, BP 89, 39801 Poligny Cedex, France.

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