Project description:The goal of this study was to use heterologous microarray hybridization to determine genomic content shared among different vesicomyid symbionts. These symbionts are closely related and can be thought of as different strains of bacteria, facilitating the use of heterologous microarray hybridization to determine genomic content. Keywords: comparative genomic hybridization Microarrays were built off the Ruthia magnifica genome and two replicate hybridizations to this organism were used as a baseline for comparisons. Genomic DNA from two other vesicomyid symbionts (Calyptogena kilmeri and C. pacifica symbionts) was also hybridized to the array with three biological replicates for each sample.
Project description:The goal of this study was to use heterologous microarray hybridization to determine genomic content shared among different vesicomyid symbionts. These symbionts are closely related and can be thought of as different strains of bacteria, facilitating the use of heterologous microarray hybridization to determine genomic content. Keywords: comparative genomic hybridization
Project description:Host-microbe interactions are virtually bidirectional, benefiting both the host and microbial sides. It is becoming increasingly recognized the influence of the microbe on many aspects of host physiology and diseases, but whether/how the host affects their symbionts is poorly characterized. Here, we reported that the host acts as a critical factor to shape the lifestyle of their symbionts in the Drosophila and bacteria model system. First, we observe that Drosophila larvae play a pivotal role in competing with pathogenic symbionts in the co-existing niche. More specifically, host larvae antagonize symbionts by deconstructing the surface slick, preventing outgrowth and antagonizing the pathogenicity of S. marcescens. Furthermore, Drosophila larvae cause the shift in the transcriptomic profile of S. marcescens, characterized with the upregulated expression of genes related to bacterial proliferation and growth and the downregulated expression of genes related to bacterial pathogenicity. More importantly, advances in bacterial single-cell RNA sequencing provide opportunities to reveal transcriptional variation, including toxic factors, across individual cells and a subpopulation clustering of isogenic bacterial populations. Finally, we found that AMPs from larvae recapitulated the response of S. marcescens to the presence of Drosophila larvae. Altogether, these findings provide an insight into the pivotal roles of the host in influencing the potential pathogens' lifecycle switching from commensalism to pathogenicity, opening the door to a better understanding of the ecological relationships between the host and microbe.
Project description:Host-microbe interactions are virtually bidirectional, benefiting both the host and microbial sides. It is becoming increasingly recognized the influence of the microbe on many aspects of host physiology and diseases, but whether/how the host affects their symbionts is poorly characterized. Here, we reported that the host acts as a critical factor to shape the lifestyle of their symbionts in the Drosophila and bacteria model system. First, we observe that Drosophila larvae play a pivotal role in competing with pathogenic symbionts in the co-existing niche. More specifically, host larvae antagonize symbionts by deconstructing the surface slick, preventing outgrowth and antagonizing the pathogenicity of S. marcescens. Furthermore, Drosophila larvae cause the shift in the transcriptomic profile of S. marcescens, characterized with the upregulated expression of genes related to bacterial proliferation and growth and the downregulated expression of genes related to bacterial pathogenicity. More importantly, advances in bacterial single-cell RNA sequencing provide opportunities to reveal transcriptional variation, including toxic factors, across individual cells and a subpopulation clustering of isogenic bacterial populations. Finally, we found that AMPs from larvae recapitulated the response of S. marcescens to the presence of Drosophila larvae. Altogether, these findings provide an insight into the pivotal roles of the host in influencing the potential pathogens' lifecycle switching from commensalism to pathogenicity, opening the door to a better understanding of the ecological relationships between the host and microbe.
Project description:Alcohol oxidases are ecologically important enzymes which facilitate a number of plant-fungal interactions. Within Ascomycota they are primarily associated with methylotrophy, as a peroxisomal alcohol oxidase (AOX) catalyzing the conversion of methanol to formaldehyde in methylotrophic yeast. In this study we demonstrate that AOX orthologs are phylogenetically conserved proteins which are common in the genomes of non-methylotrophic, plant-associating fungi. Additionally, AOX orthologs are highly expressed during infection in a range of diverse pathosystems. To study the role of AOX in plant colonization, AOX knockout mutants were generated in the broad host range pathogen Sclerotinia sclerotiorum. Disease assays in soybean showed that these mutants had a significant virulence defect as evidenced by markedly reduced stem lesions and mortality rates. Chemical genomics suggest that SsAOX may function as an aromatic alcohol oxidase, and growth assays demonstrate that ΔSsAOX is incapable of properly utilizing plant extract as a nutrient source. Profiling of known aromatic alcohols point towards the monolignol coniferyl alcohol (CoA) as a possible substrate for SsAOX. As CoA and other monolignols are ubiquitous among land plants, the presence of highly conserved AOX orthologs throughout Ascomycota imply that this is a broadly conserved protein used by ascomycete fungi during plant colonization.