Project description:Plant response to insect feeding appears to be highly specific with regard to the organisms in the system. Here, we report on the interaction between grapevine Vitis vinifera plants and a phloem-feeding insect pest, the vine mealybug Planococcus ficus. Plants were exposed to P. ficus for periods of 6 hours and 96 hours, after which they were analysed for gene expression levels using microarrays and quantitative real-time PCR (qPCR). Both methods showed that grapevine displayed only a minimal response to mealybug feeding at the transcript level at both time periods. Intermediate grapevine exposure times (24, 48 and 72 hours) to P. ficus feeding were investigated using qPCR analysis of ten additional genes associated with known plant defense responses. Results showed that only a single gene, pathogenesis-related protein 1, was differentially expressed after 48 hours of mealybug feeding. During the course of mealybug feeding, however, a number of other genes were significantly up- or down-regulated at certain time points. Thus, it appears as if grapevine responds minimally to feeding by P. ficus as well as within a very narrow time period. The relative lack of grapevine plant defense mechanisms may be a result of the feeding strategies of mealybugs.
Project description:Plant response to insect feeding appears to be highly specific with regard to the organisms in the system. Here, we report on the interaction between grapevine Vitis vinifera plants and a phloem-feeding insect pest, the vine mealybug Planococcus ficus. Plants were exposed to P. ficus for periods of 6 hours and 96 hours, after which they were analysed for gene expression levels using microarrays and quantitative real-time PCR (qPCR). Both methods showed that grapevine displayed only a minimal response to mealybug feeding at the transcript level at both time periods. Intermediate grapevine exposure times (24, 48 and 72 hours) to P. ficus feeding were investigated using qPCR analysis of ten additional genes associated with known plant defense responses. Results showed that only a single gene, pathogenesis-related protein 1, was differentially expressed after 48 hours of mealybug feeding. During the course of mealybug feeding, however, a number of other genes were significantly up- or down-regulated at certain time points. Thus, it appears as if grapevine responds minimally to feeding by P. ficus as well as within a very narrow time period. The relative lack of grapevine plant defense mechanisms may be a result of the feeding strategies of mealybugs. Eight samples were analysed. Two replicates each were included for each treatment (6 hour and 96 hour feeding), resulting in four samples. Two control replicates were included for each treatment (6 hour and 96 hour feeding controls), resulting in a further four samples.
Project description:Persistent mucosal inflammation and microbial infection are characteristic of Chronic Rhinosinusitis (CRS). Though mucosal microbiota dysbiosis is a characteristic feature of other chronic inflammatory diseases, the relationship between sinus microbiota composition and CRS is unknown. Here we demonstrate, using comparative microbiome profiling of a cohort of CRS patients and healthy subjects, that the sinus microbiota of CRS patients exhibit significantly reduced bacterial diversity. Characteristic of this community collapse is the depletion of multiple, phylogenetically distinct, Lactic Acid Bacteria and the concomitant increase in relative abundance of a single species, Corynebacterium tuberculostearicum. Recapitulating the conditions observed in our human cohort in a murine model confirmed the pathogenic potential of C. tuberculostearicum and the critical necessity for a replete mucosal microbiota to protect against this species. Moreover, we provide evidence that Lactobacillus sakei, identified from our comparative microbiome analyses as a potentially protective species, affords defense against C. tuberculostearicum sinus infection, even in the context of a depleted sinus bacterial community. These studies demonstrate that sinus mucosal health is highly dependent on the composition of the resident microbiota, and identifies a new sino-pathogen and a strong bacterial candidate for therapeutic intervention. A total of 14 samples were profiled for microbiome composition: 7 from non-sinusitis patients, and 7 from patients with clinically diagnosed chronic sinusitis.
Project description:P. citri mealybugs cause severe damage in citrus production. We aimed at indentifying genes involved in mealybug sex pheromone biosynthesis. As these should be expressed in virgin females and are presumably downregulated after mating, we looked at genes differentially expressed between mated and virgin females.
Project description:Persistent mucosal inflammation and microbial infection are characteristic of Chronic Rhinosinusitis (CRS). Though mucosal microbiota dysbiosis is a characteristic feature of other chronic inflammatory diseases, the relationship between sinus microbiota composition and CRS is unknown. Here we demonstrate, using comparative microbiome profiling of a cohort of CRS patients and healthy subjects, that the sinus microbiota of CRS patients exhibit significantly reduced bacterial diversity. Characteristic of this community collapse is the depletion of multiple, phylogenetically distinct, Lactic Acid Bacteria and the concomitant increase in relative abundance of a single species, Corynebacterium tuberculostearicum. Recapitulating the conditions observed in our human cohort in a murine model confirmed the pathogenic potential of C. tuberculostearicum and the critical necessity for a replete mucosal microbiota to protect against this species. Moreover, we provide evidence that Lactobacillus sakei, identified from our comparative microbiome analyses as a potentially protective species, affords defense against C. tuberculostearicum sinus infection, even in the context of a depleted sinus bacterial community. These studies demonstrate that sinus mucosal health is highly dependent on the composition of the resident microbiota, and identifies a new sino-pathogen and a strong bacterial candidate for therapeutic intervention.
Project description:Host pathways mediating changes in immune states elicited by intestinal microbial colonization are incompletely characterized. Here we describe alterations of the host immune state induced by colonization of germ-free zebrafish larvae with an intestinal microbial community or single bacterial species. We show that microbiota-induced changes in intestinal leukocyte subsets and whole-body host gene expression are dependent on the innate immune adaptor gene myd88. Similar patterns of gene expression are elicited by colonization with conventional microbiome, as well as mono-colonization with two different zebrafish commensal bacterial strains. By studying loss-of-function myd88 mutants, we find that colonization suppresses Myd88 at the mRNA level. Tlr2 is essential for microbiota-induced effects on myd88 transcription and intestinal immune cell composition.
Project description:Here we report a direct tRNA sequencing protocol and software to simultaneously examine the composition and biological activity of naturally occurring microbial communities. Our analysis of mouse gut microbiome with tRNA-seq and 16S ribosomal RNA gene amplicons revealed comparable microbial community structures, and additional physiological insights into the microbiome through tRNA abundance and modifications.