Luminescence-based systems for detection of bacteria in the environment

James Ivor Prosser, Kenneth Stuart Killham, Lesley Anne Glover, Elizabeth-Ann Simpson Rattray

Research output: Contribution to journalLiterature review

48 Citations (Scopus)

Abstract

The development of techniques for detection and tracking of microorganisms in natural environments has been accelerated by the requirement for assessment of the risks associated with environmental release of genetically engineered microbial inocula. Molecular marker systems are particularly appropriate for such studies and luminescence-based markers have the broadest range of applications, involving the introduction of prokaryotic (lux) or eukaryotic (luc) genes for the enzyme luciferase.

Lux or luc genes can be detected on the basis of unique DNA sequences by gene probing and PCR amplification, but the major advantage of luminescence-based systems is the ability to detect light emitted by marked organisms or by luciferase activity in cell-free extracts. Luminescent colonies can be detected by eye, providing distinction from colonies of indigenous organisms, and the sensitivity of plate counting can be increased greatly by CCD imaging. Single cells or microcolonies of luminescent organisms can also be detected in environmental samples by CCD image-enhanced microscopy, facilitating study of their spatial distribution. The metabolic activity of luminescence-marked populations can be quantified by luminometry and does not require extraction of cells or laboratory growth. Metabolic activity, and potential activity, of marked organisms therefore can be measured during colonization of soil particles and plant material in real time without disturbing the colonization process.

In comparison with traditional activity techniques, luminometry provides significant increases in sensitivity, accuracy, and, most importantly, selectivity, as activity can be measured in the presence of indigenous microbial communities. The sensitivity, speed, and convenience of luminescence measurements make this a powerful technique that is being applied to the study of an increasingly wide range of ecological problems. These include microbial survival and recovery, microbial predation, plant pathogenicity, phylloplane and rhizosphere colonization and reporting of gene expression in environmental samples.

Original languageEnglish
Pages (from-to)157-183
Number of pages27
JournalCritical Reviews in Biotechnology
Volume16
Issue number2
Publication statusPublished - 1996

Keywords

  • luminescence-based microbial detection
  • detection of genetically engineered microorganisms in the environment
  • molecular-based microbial marker systems
  • survival and recovery of microorganisms in natural environments
  • measurement of microbial activity in natural environments
  • microbial predation
  • microbial colonization of the phylloplane and rhizosphere
  • GENETICALLY ENGINEERED MICROORGANISMS
  • LOCUS CONTROLLING EXPRESSION
  • FIREFLY LUCIFERASE GENE
  • AMINO-ACID SEQUENCE
  • VIBRIO-HARVEYI
  • NUCLEOTIDE-SEQUENCE
  • STRUCTURAL IDENTIFICATION
  • PSEUDOMONAS-FLUORESCENS
  • RHIZOBIUM-MELILOTI
  • ERWINIA-CAROTOVORA

Cite this

Prosser, J. I., Killham, K. S., Glover, L. A., & Rattray, E-A. S. (1996). Luminescence-based systems for detection of bacteria in the environment. Critical Reviews in Biotechnology, 16(2), 157-183.