Design of novel primer sets for easy detection of Ruegeria species from seawater

Biosci Biotechnol Biochem. 2020 Apr;84(4):854-864. doi: 10.1080/09168451.2019.1700776. Epub 2019 Dec 9.

Abstract

Some coral-associated bacteria show protective roles for corals against pathogens. However, the distribution of coral-protecting bacteria in seawater is not well known. In addition, compared with the methods for investigating coral pathogens, few methods have been developed to detect coral-protecting bacteria. Here we prepared a simple method for detecting Ruegeria spp., some strains of which inhibit growth of the coral pathogen Vibrio coralliilyticus. We successfully obtained two Ruegeria-targeting primer sets through in silico and in vitro screening. The primer sets r38F-r30R and r445F-r446R, in addition to the newly designed universal primer set U357'F-U515'R, were evaluated in vitro using environmental DNA extracted from seawater collected in Osaka. These methods and primers should contribute to revealing the distribution of Ruegeria spp. in marine environments.

Keywords: Ruegeria; coral-associated bacteria; in silico screening; real-time PCR; seawater.

MeSH terms

  • Animals
  • Anthozoa / microbiology
  • DNA Primers*
  • Electrophoresis, Polyacrylamide Gel
  • Polymerase Chain Reaction
  • Rhodobacteraceae / genetics*
  • Rhodobacteraceae / isolation & purification*
  • Seawater*

Substances

  • DNA Primers