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.
The insect pool available for carrion visitation and colonisation varies with geographical areas, hence the need to build a comprehensive database wherever such data could be used in forensic investigations. However, most of the geographic records on carrion-related insects are from short-term seasonal studies. Here, we provide the year-round taxonomic composition for the dominant ecosystem of the Maritime lowland ecological region that borders the Gulf of St. Lawrence, Canada, and we examine how this composition is subjected to natural (seasonal) as well as anthropogenic changes. Fresh pig carcasses, used here as human surrogates, were exposed recurrently throughout the whole annual period when carrion-related insects are active in forests and adjacent areas subjected to human-induced land cover changes from agricultural practices. A total of 130 necrophageous and predacious insect species representing 2 orders, 18 families and 75 genera were recovered from carcasses. Abundant fly species were able to visit and/or colonise carcasses exposed in both forests and agricultural fields but the species involved varied throughout the year. Conversely, the complex of abundant coleopterans found on carcasses remained stable throughout the year but differed between forests and agricultural fields. Considering the seasonal and anthropogenic changes that were observed in the complex of carrion-related insects, we stressed that inference on the taxonomic composition in relation to minimum postmortem interval should be restricted to a specific habitat and time of the year. These results also have methodological implications, suggesting that the experimental designs of forensic studies in temperate areas require adjustments to permit robust estimations of minimum postmortem intervals from the insect fauna associated with carcasses.
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