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Research Article Open access CC BY 3.0

Isolation and Molecular Characterization of Lactic Acid Bacteria Isolated from Fresh Fruits and Vegetables Using Nested PCR Analysis

E. C. Emerenini, O. R. Afolabi, P. I. Okolie, A. K. Akintokun

Microbiology Research Journal International · pp. 368–377 · Published 16 Jun 2013

10.9734/BMRJ/2013/2520

Abstract

Aims: The study investigated the diversity and identities of Lactic Acid Bacteria (LAB) isolated from different fresh fruits and vegetables using Molecular Nested PCR analysis with the view of identifying LAB with anti-microbial potentials. Study Design: Nested PCR approach was used in this study employing universal 16S rRNA gene primers in the first round PCR and LAB specific Primers in the second round PCR with the view of generating specific Nested PCR products for the LAB diversity present in the samples. Place and Duration of Study: Biotechnology Centre of Federal University of Agriculture, Abeokuta, Ogun State, Nigeria, between January 2011 and February 2012. Methodology: Forty Gram positive, catalase negative strains of LAB were isolated from fresh fruits and vegetables on Man Rogosa and Sharpe agar (Lab M) using streaking method. Standard molecular methods were used for DNA extraction (Norgen Biotek kit method, Canada), Polymerase Chain Reaction (PCR) Amplification, Electrophoresis, Purification and Sequencing of generated Nested PCR products (Macrogen Inc., USA). Results: The partial sequences obtained were deposited in the database of National Centre for Biotechnology Information (NCBI). Isolates were identified based upon the sequences as Weissella cibaria (5 isolates, 27.78%), Weissella kimchi (5, 27.78%), Weissella paramensenteroides (3, 16.67%), Lactobacillus plantarum (2, 11.11%), Pediococcus pentosaceus (2, 11.11%) and Lactobacillus pentosus (1, 5.56%) from fresh vegetable; while Weissella cibaria (4, 18.18%), Weissella confusa (3, 13.64%), Leuconostoc paramensenteroides (1, 4.55%), Lactobacillus plantarum (8, 36.36%), Lactobacillus paraplantarum (1, 4.55%) and Lactobacillus pentosus (1, 4.55%) were identified from fresh fruits. Conclusion: This study shows that potentially LAB can be quickly and holistically characterized by molecular methods to specie level by nested PCR analysis of the bacteria isolate genomic DNA using universal 16S rRNA primers and LAB specific primer.

Nested PCR Molecular characterization 16S rRNA gene Lactic acid bacteria.

References (18)

  1. 1 Polyphasic taxonomy, a consensus approach to bacterial systematics [DOI]
  2. 2 Phenotypic and molecular identification and clustering of lactic acid bacteria and yeasts from wheat (species Triticum durum and Triticum aestivum) sourdoughs of Southern Italy [DOI]
  3. 3 Lactic acid bacteria from fresh fruit and vegetables as biocontrol agents of phytopathogenic bacteria and fungi. [DOI]
  4. 4 Design and Evaluation of PCR Primers for Analysis of Bacterial Populations in Wine by Denaturing Gradient Gel Electrophoresis [DOI]
  5. 5 Isolation and identification of lactic acid bacteria from soil using an enrichment procedure [DOI]
  6. 6 Lactobacillus casei, dominant species in naturally fermented Sicilian green olives [DOI]
  7. 7 Safety of industrial lactic acid bacteria [DOI]
  8. 8 Microflora identification of the Bulgarian cereal-based fermented beverage boza [DOI]
  9. 9 Characterization of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation [DOI]
  10. 10 Biochemical characterization of lactic acid bacteria isolated from spontaneous fermentation of ‘Almagro’ eggplants [DOI]
  11. 11 Identification of the lactic acid bacteria in Kimchi according to initial and over-ripened fermentation using PCR and 16S rRNA gene sequence analysis [DOI]
  12. 12 Isolation, characterization and identification of lactic acid bacteria involved in traditional fermentation of borde, an Ethiopian cereal beverage [DOI]
  13. 13 SCREENING AND IDENTIFICATION OF LACTIC ACID BACTERIA PRODUCING ANTIMICROBIAL COMPOUNDS FROM PIG GASTROINTESTINAL TRACTS
  14. 14 Isolation and Applications of One Strain of Lactobacillus paraplantarum from Tea Leaves (Camellia sinensis) [DOI]
  15. 15 Identifications of Predominant Bacterial Isolates from the Fermenting Kimchi Using ITS-PCR and Partial 16S rDNA Sequence Analyses
  16. 16 Cloning and characterization of a gene encoding phosphoketolase in a Lactobacillus paraplantarum isolated from Kimchi.
  17. 17 Microbial Enzymes Associated with Fresh-cut Produce [DOI]
  18. 18 Isolation and Identification of Epiphytic Lactic Acid Bacteria from Guinea Grass (Panicum maximum) [DOI]

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