Characterization of the Bacterial Community Naturally Present on Commercially Grown Basil Leaves: Evaluation of Sample Preparation Prior to Culture-Independent Techniques

Int J Environ Res Public Health. 2015 Aug 21;12(8):10171-97. doi: 10.3390/ijerph120810171.

Abstract

Fresh herbs such as basil constitute an important food commodity worldwide. Basil provides considerable culinary and health benefits, but has also been implicated in foodborne illnesses. The naturally occurring bacterial community on basil leaves is currently unknown, so the epiphytic bacterial community was investigated using the culture-independent techniques denaturing gradient gel electrophoresis (DGGE) and next-generation sequencing (NGS). Sample preparation had a major influence on the results from DGGE and NGS: Novosphingobium was the dominant genus for three different basil batches obtained by maceration of basil leaves, while washing of the leaves yielded lower numbers but more variable dominant bacterial genera including Klebsiella, Pantoea, Flavobacterium, Sphingobacterium and Pseudomonas. During storage of basil, bacterial growth and shifts in the bacterial community were observed with DGGE and NGS. Spoilage was not associated with specific bacterial groups and presumably caused by physiological tissue deterioration and visual defects, rather than by bacterial growth.

Keywords: 16S rRNA; DGGE; Novosphingobium; fresh herbs; next-generation sequencing NGS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / classification
  • Bacteria / genetics
  • Bacteria / isolation & purification*
  • DNA, Bacterial / genetics
  • Denaturing Gradient Gel Electrophoresis
  • Food Microbiology*
  • Food Storage*
  • Microbiota*
  • Ocimum basilicum / microbiology*
  • Plant Leaves / microbiology
  • Polymerase Chain Reaction
  • RNA, Ribosomal, 16S / genetics
  • Temperature

Substances

  • DNA, Bacterial
  • RNA, Ribosomal, 16S

Associated data

  • SRA/PRJNA288639