BackgroundThe mountain pine beetle (MPB, Dendroctonus ponderosae Hopkins) is a highly destructive pest of pine forests in western North America. During flight to a new host tree and initiation of feeding, mountain pine beetles release aggregation pheromones. The biosynthetic pathways of these pheromones are sex-specific and localized in the midgut and fat body, but the enzymes involved have not all been identified or characterized.ResultsWe used a comparative RNA-Seq analysis between fed and unfed male and female MPB midguts and fat bodies to identify candidate genes involved in pheromone biosynthesis. The 13,407 potentially unique transcripts showed clear separation based on feeding state and gender. Gene co-expression network construction and examination using petal identified gene groups that were tightly connected. This, as well as other co-expression and gene ontology analyses, identified all four known pheromone biosynthetic genes, confirmed the tentative identification of four others from a previous study, and suggested nine novel candidates. One cytochrome P450 monooxygenase, CYP6DE3, identified as a possible exo-brevicomin-biosynthetic enzyme in this study, was functionally characterized and likely is involved in resin detoxification rather than pheromone biosynthesis.ConclusionsOur analysis supported previously characterized pheromone-biosynthetic genes involved in exo-brevicomin and frontalin biosynthesis and identified a number of candidate cytochrome P450 monooxygenases and a putative cyclase for further studies. Functional analyses of CYP6DE3 suggest its role in resin detoxification and underscore the limitation of using high-throughput data to tentatively identify candidate genes. Further functional analyses of candidate genes found in this study should lead to the full characterization of MPB pheromone biosynthetic pathways and the identification of molecular targets for possible pest management strategies.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-017-3696-4) contains supplementary material, which is available to authorized users.
Increases in the growth rate of plants and microbes in the Mojave Desert in response to predicted increases in precipitation and CO 2 due to global climate change may induce nutrient limitations. This study was designed to measure the pool of potentially bioavailable nutrients in soils of the Mojave Desert. Soils were collected from shrub and interspace microsites and then subjected to amendment with buffered solutions of an excess of various enzymes. The products of each enzyme reaction were then measured and the maximum quantity of hydrolyzable substrates was calculated. In interspace and shrub microsite soils, respectively, 14.5 and 9.7% of the organic C in the form cellulose, 60.0-97.8% and 61.2-100.0% of the organic N in the form protein, and 44.0 and 57.5% of the organic P was hydrolyzable. There were significant differences between microsites for hydrolyzable substrate using all enzyme amendments, except protease. We propose that accumulations of hydrolyzable organic C, N, and P in the Mojave Desert could be a result of the persistently dry soil conditions often found in desert ecosystems and the immobilization of enzymes, which may result in low diffusivity of soil substrates and enzymes and, accordingly, little degradation of organic C, N, and P. Alternatively, rapid nutrient cycling and immobilization by soil microorganisms could account for accumulations of organic C, N, and P. Further refinement of the methods used in this study could lead to a valuable tool for the assessment of potential bioavailability of nutrients in a variety of soils.
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