Application of relative real-time PCR to detect differential expression of virulence genes in Vibrio anguillarum under standard and stressed growth conditions

In this study, we aimed to understand whether abiotic factors affect the expression of virulence genes in Vibrio anguillarum. We observed the in vitro responses of two Mediterranean strains of V. anguillarum to temperature, NaCl and iron concentration changes. We monitored growth performance and gene transcription levels by comparing the results obtained under stressed conditions (temperatures of 5 °C, 15 °C and 37 °C; NaCl concentrations of 3% and 5%; and iron depletion and excess) with those obtained under standard growth conditions (25 °C, 1.5% NaCl and 0.6 ?m of iron). The results showed that the strains respond differently. The strain 975/I was most strongly affected by conditions of 15 °C and iron depletion; these conditions induced increased transcription levels of empA, angR and fatA. Growth of the strain 17/I was inhibited at 15 °C and in iron depletion conditions; this strain also showed dramatic changes in the transcription levels of toxR and tonB2 under increased NaCl concentrations. These results demonstrate that environmental stress affects the expression of virulence genes in V. anguillarum that have implications for the competitiveness, stress tolerance and the ability of V. anguillarum to cause infection. © 2013 John Wiley & Sons Ltd.