Summary
Morchella rufobrunnea is a saprobic edible mushroom, found in a range of ecological niches, indicating nutritional adjustment to different habitats and possible interaction with soil prokaryotic microbiome (SPM). Using the 16S rRNA gene, we examined the SPM of M. rufobrunnea that appeared in a natural habitat in Northern Israel. Three sample types were included: bare soil without mushroom, soil beneath young mushroom initials and soil beneath the mature fruiting body. Morchella rufobrunnea developmental stage was significantly associated with changes in bacterial populations (PERMANOVA, p < 0.0005). Indicator analysis with point‐biserial correlation coefficient found 180 operational taxonomic units (OTU) uniquely associated with distinct stages of development. The Functional Annotation of Prokaryotic Taxonomy (FAPROTAX) database helped to infer ecological roles for indicator OTU. The functional ecological progression begins with establishment of a photoautotrophic N‐fixing bacterial mat on bare soil. Pioneer heterotrophs including oligotrophs, acidifying nutrient mobilizers and nitrifiers are congruent with appearance of young M. rufobrunnea initials. Under the mature fruiting body, the population changed to saprobes, organic‐N degraders, denitrifiers, insect endosymbionts and fungal antagonists. Based on this work, M. rufobrunnea may be able to influence SPM and change the soil nutritional profile.
A sensitive, high-throughput, and costeffective method for screening bacterial pathogens in the environment was developed. A variety of environmental samples, including aerosols, soil of various types (sand, sand/clay mix, and clay), wastewater, and vegetable surface (modeled by tomato), were concomitantly spiked with Salmonella enterica and/or Pseudomonas aeruginosa to determine recovery rates and limits of detection. The various matrices were first enriched with a general pre-enrichment broth in a dilution series and then enumerated by most probable number (MPN) estimation using quantitative PCR for rapid screening of amplicon presence. Soil and aerosols were then tested in non-spiked environmental samples, as these matrices are prone to large experimental variation. Limit of detection in the various soil types was 1-3 colony-forming units (CFU) g −1 ; on vegetable surface, 5 CFU per tomato; in treated wastewater, 5 CFU L −1 ; and in aerosols, >300 CFU mL −1 . Our method accurately identified S. enterica in non-spiked environmental soil samples within a day, while traditional methods took 4 to 5 days and required sorting through biochemically and morphologically similar species. Likewise, our method successfully identified P. aeruginosa in non-spiked aerosols generated by a domestic wastewater treatment system. The obtained results suggest that the developed method presents a broad approach for the rapid, efficient, and reliable detection of relatively low densities of pathogenic organisms in challenging environmental samples.
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