This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Blaiotta, G.
Right arrow Articles by Villani, F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Blaiotta, G.
Right arrow Articles by Villani, F.
Agricola
Right arrow Articles by Blaiotta, G.
Right arrow Articles by Villani, F.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, January 2008, p. 208-215, Vol. 74, No. 1
0099-2240/08/$08.00+0     doi:10.1128/AEM.01711-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Lactobacillus Strain Diversity Based on Partial hsp60 Gene Sequences and Design of PCR-Restriction Fragment Length Polymorphism Assays for Species Identification and Differentiation{triangledown} ,{dagger}

Giuseppe Blaiotta,1* Vincenzina Fusco,1 Danilo Ercolini,2 Maria Aponte,1 Olimpia Pepe,1 and Francesco Villani1

Department of Food Science, School of Agriculture,1 School of Biotechnological Sciences, The University of Naples Federico II, via Università 100, 80055 Portici, Italy2

Received 25 July 2007/ Accepted 30 October 2007

A phylogenetic tree showing diversities among 116 partial (499-bp) Lactobacillus hsp60 (groEL, encoding a 60-kDa heat shock protein) nucleotide sequences was obtained and compared to those previously described for 16S rRNA and tuf gene sequences. The topology of the tree produced in this study showed a Lactobacillus species distribution similar, but not identical, to those previously reported. However, according to the most recent systematic studies, a clear differentiation of 43 single-species clusters was detected/identified among the sequences analyzed. The slightly higher variability of the hsp60 nucleotide sequences than of the 16S rRNA sequences offers better opportunities to design or develop molecular assays allowing identification and differentiation of either distant or very closely related Lactobacillus species. Therefore, our results suggest that hsp60 can be considered an excellent molecular marker for inferring the taxonomy and phylogeny of members of the genus Lactobacillus and that the chosen primers can be used in a simple PCR procedure allowing the direct sequencing of the hsp60 fragments. Moreover, in this study we performed a computer-aided restriction endonuclease analysis of all 499-bp hsp60 partial sequences and we showed that the PCR-restriction fragment length polymorphism (RFLP) patterns obtainable by using both endonucleases AluI and TacI (in separate reactions) can allow identification and differentiation of all 43 Lactobacillus species considered, with the exception of the pair L. plantarum/L. pentosus. However, the latter species can be differentiated by further analysis with Sau3AI or MseI. The hsp60 PCR-RFLP approach was efficiently applied to identify and to differentiate a total of 110 wild Lactobacillus strains (including closely related species, such as L. casei and L. rhamnosus or L. plantarum and L. pentosus) isolated from cheese and dry-fermented sausages.

* Corresponding author. Mailing address: Dipartimento di Scienza degli Alimenti, Università degli Studi di Napoli Federico II, Via Università 100, 80055 Portici (Naples), Italy. Phone: 39-081-2539451. Fax: 39-081-2539407. E-mail: blaiotta{at}unina.it

{triangledown} Published ahead of print on 9 November 2007.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, January 2008, p. 208-215, Vol. 74, No. 1
0099-2240/08/$08.00+0     doi:10.1128/AEM.01711-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Glazunova, O. O., Raoult, D., Roux, V. (2009). Partial sequence comparison of the rpoB, sodA, groEL and gyrB genes within the genus Streptococcus. Int. J. Syst. Evol. Microbiol. 59: 2317-2322 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»