title: Transcriptional activity of antifungal metabolite-encoding genes phlD and hcnBC in Pseudomonas spp. using qRT-PCR.
abstract: Production of 2,4-diacetylphloroglucinol (2,4-DAPG) and hydrogen cyanide (HCN) by Pseudomonas spp. shows great potential for controlling soilborne plant pathogens. However, little is known about the transcriptional activity of phl and hcn genes encoding 2,4-DAPG and HCN, respectively. To progress toward a better understanding of what triggers phl and hcn expression under rhizosphere conditions, novel PCR primers and TaqMan probes were designed to monitor relative phlD and hcnBC expression in quantitative real time-PCR assays. Transcriptional activity of phlD and hcnBC was studied in time-course confrontational assays using combinations of Pseudomonas spp. isolated in this study: LBUM300 (producing 2,4-DAPG and HCN) and LBUM647 (producing HCN only); pathogens Phytophthora cactorum and Verticillium dahliae; and solid growth media King's B medium and potato dextrose agar. In correlation with the antagonistic activity observed, expression of phlD and hcnBC and production of 2,4-DAPG was detected throughout the 14-day course of the experiment in LBUM300 on both media, while hcnBC expression diminished to undetectable levels in LBUM647. In LBUM300 expression of phlD and hcnBC significantly changed over time and was also influenced by the presence of pathogen and growth media following time-dependent responses.
authors: Department of Biology, Université de Moncton, Moncton, New Brunswick, Canada.
authors: Mélanie M
authors: MM
authors: Paulin
authors: Amy
authors: A
authors: Novinscak
authors: Marc
authors: M
authors: St-Arnaud
authors: Claudia
authors: C
authors: Goyer
authors: Nadine J
authors: NJ
authors: DeCoste
authors: Jean-Pierre
authors: JP
authors: Privé
authors: Josée
authors: J
authors: Owen
authors: Martin
authors: M
authors: Filion
mesh_terms: D000898:Antibiosis; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005798:Genes, Bacterial; D010696:Phloroglucinol / Q000031:analogs & derivatives / Q000378:metabolism; D010838:Phytophthora / Q000502:physiology; D011549:Pseudomonas / Q000235:genetics* / Q000378:metabolism; D012329:RNA, Bacterial / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012988:Soil Microbiology; D014158:Transcription, Genetic; D020172:Verticillium / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; C096183:PhlD protein, Pseudomonas; D012329:RNA, Bacterial; C059817:2,4-diacetylphloroglucinol; D010696:Phloroglucinol