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Microbial biosynthetic gene clusters (BGCs) encoding secondary metabolites are thought to impact a plethora of biologically mediated environmental processes, yet their discovery and functional characterization in natural microbiomes remains challenging. Here we describe deep long-read sequencing and assembly of metagenomes from biological soil crusts, a group of soil communities that are rich in BGCs. Taking advantage of the unusually long assemblies produced by this approach, we recovered nearly 3,000 BGCs for analysis, including 712 full-length BGCs. Functional exploration through metatranscriptome analysis of a 3-day wetting experiment uncovered phylum-specific BGC expression upon activation from dormancy, elucidating distinct roles and complex phylogenetic and temporal dynamics in wetting processes. For example, a pronounced increase in BGC transcription occurs at night primarily in cyanobacteria, implicating BGCs in nutrient scavenging roles and niche competition. Taken together, our results demonstrate that long-read metagenomic sequencing combined with metatranscriptomic analysis provides a direct view into the functional dynamics of BGCs in environmental processes and suggests a central role of secondary metabolites in maintaining phylogenetically conserved niches within biocrusts. Marc Van Goethem et al. combine short- and long-read sequencing from biocrust samples to report nearly 3,000 biosynthetic gene clusters (BGCs) encoding microbial secondary metabolites. Their results demonstrate the advantage of integrated metatranscriptomic and long-read metagenomic sequencing in analyzing BGCs and provides insight into the role of secondary metabolites in maintaining phylogenetic niches in biocrusts. [ABSTRACT FROM AUTHOR]

Hydrocarbon degradation is usually measured in laboratories under controlled conditions to establish the likely efficacy of a bioremediation process in the field. The present study used greenhouse-based bioremediation to investigate the effects of natural attenuation (NA) and necrophytoremediation (addition of pea straw (PS)) on hydrocarbon degradation, toxicity and the associated bacterial community structure and composition in diesel-contaminated soil. A significant reduction in total petroleum hydrocarbon (TPH) concentration was detected in both treatments; however, PS-treated soil showed more rapid degradation (87%) after 5 months together with a significant reduction in soil toxicity (EC50 = 91 mg diesel/kg). Quantitative PCR analysis revealed an increase in the number of 16S rRNA and alkB genes in the PS-amended soil. Substantial shifts in soil bacterial community were observed during the bioremediation, including an increased abundance of numerous hydrocarbon-degrading bacteria. The bacterial community shifted from dominance by Alphaproteobacteria and Gammaproteobacteria in the original soil to Actinobacteria during bioremediation. The dominance of two genera of bacteria, Sphingobacteria and Betaproteobacteria, in both NA- and PS-treated soil demonstrated changes occurring within the soil bacterial community through the incubation period. Additionally, pea straw itself was found to harbour a diverse hydrocarbonoclastic community including Luteimonas, Achromobacter, Sphingomonas, Rhodococcus and Microbacterium. At the end of the experiment, PS-amended soil exhibited reduced ecotoxicity and increased bacterial diversity as compared with the NA-treated soil. These findings suggest the rapid growth of species stimulated by the bioremediation treatment and strong selection for bacteria capable of degrading petroleum hydrocarbons during necrophytoremediation. [ABSTRACT FROM AUTHOR]

Footrot disease due to Nectria haematococca (anamorph Fusarium solani f.sp. pisi) is an economically important disease of peas globally. However, our ability to predict accurately the likelihood of footrot infections is limited because there is no method to determine inoculum density prior to planting. In this research, a PCR-based assay was developed to quantify the pea pathogenicity gene ( PEP3), exclusive to highly pathogenic forms of N. haematococca, from DNA extracted from agricultural field soils. The applicability of using quantitative PCR (qPCR) to measure this gene in soil was validated, and the relationship between PEP3 gene numbers and footrot disease was also studied. Results showed that the quantitative assay is both efficient and specific; amplification efficiency of the Q-PCR assay for the PEP3 gene was 92%. Gene copy numbers were shown to vary significantly ( P = 0.01) between fields, and were positively correlated to the number of spores of pathogenic N. haematococca, and to footrot disease. PEP3 numbers of up to 100 g soil constituted a threshold number for infection-potentially capable of causing economically significant pea footrot disease. The density of virulent N. haematococca in soil fields capable of causing footrot disease could be determined with a high degree of accuracy, with this assay. It offers the opportunity for prediction of pea footrot infections in agricultural soils prior to cultivation. [ABSTRACT FROM AUTHOR]