Project description:Native host plant insect resistance in the maize inbred line Mp708 was developed by traditional plant breeding. Resistant Mp708 thwarts feeding by fall armyworm (Spodoptera frugiperda [J.E. Smith]; Lepidoptera: Noctuidae), numerous other lepidopteran pests, and the coleopteran western corn rootworm. This broad resistance makes it an excellent model for studying native host plant resistance mechanisms. In response to caterpillar feeding, Mp708 rapidly mobilizes Mir1-CP, a unique cysteine protease that appears to translocate from roots to the maize midwhorl where it accumulates. This accumulation correlates with a significant reduction in caterpillar growth resulting from diminished food utilization. In addition, the peritrophic membrane (PM) that surrounds the food bolus in the mudgut (MG) is severely damaged in caterpillars fed on sweet corn callus transformed to express the gene encoding Mir1-CP or on midwhorl tissue from resistant Mp708 maize. Functions of the PM include assisting digestion and protecting the epithelium of the caterpillar MG from physical and chemical damage. Consequently, the reduced growth of caterpillars that feed on Mp708 is probably due to the action of Mir1-CP on PM physiology. In fact, previous in vitro studies indicated that Mir1-CP was capable of permeabilizing the PM. The present study used both targeted (qRT-PCR) and global (mRNA-seq) transcriptome analyses to explore the effect of eating Mir1-CP expressing Mp708 maize on abundance of transcripts in the MG of fall armyworm larvae in comparison to MGs from larvae fed on susceptible Tx601 maize that does not express Mir1-CP. Expression of genes encoding proteins involved in PM production is upregulated in MGs from fall armyworm fed on Mp708. Also, several digestive enzymes (endopeptidases, aminopeptidases, lipases, amylase) were more highly expressed in MGs from larvae fed on Mp708 than MGs from larvae fed on Tx601. Impaired growth of larvae fed on Mp708 probably results from metabolic costs associated with higher production of PM constituents and digestive enzymes in a compensatory attempt to maintain MG function.
Project description:Given the criticle role of gut bacteria involve in number of diseases, the gut microbiota from young and aged people were estimated using 16s rRNA next-generation sequencing. This study will benefit to identify the role of gut bacteria on the pathegenic mechasim of aging relative diseases.
Project description:Native host plant insect resistance in the maize inbred line Mp708 was developed by traditional plant breeding. Resistant Mp708 thwarts feeding by fall armyworm (Spodoptera frugiperda [J.E. Smith]; Lepidoptera: Noctuidae), numerous other lepidopteran pests, and the coleopteran western corn rootworm. This broad resistance makes it an excellent model for studying native host plant resistance mechanisms. In response to caterpillar feeding, Mp708 rapidly mobilizes Mir1-CP, a unique cysteine protease that appears to translocate from roots to the maize midwhorl where it accumulates. This accumulation correlates with a significant reduction in caterpillar growth resulting from diminished food utilization. In addition, the peritrophic membrane (PM) that surrounds the food bolus in the mudgut (MG) is severely damaged in caterpillars fed on sweet corn callus transformed to express the gene encoding Mir1-CP or on midwhorl tissue from resistant Mp708 maize. Functions of the PM include assisting digestion and protecting the epithelium of the caterpillar MG from physical and chemical damage. Consequently, the reduced growth of caterpillars that feed on Mp708 is probably due to the action of Mir1-CP on PM physiology. In fact, previous in vitro studies indicated that Mir1-CP was capable of permeabilizing the PM. The present study used both targeted (qRT-PCR) and global (mRNA-seq) transcriptome analyses to explore the effect of eating Mir1-CP expressing Mp708 maize on abundance of transcripts in the MG of fall armyworm larvae in comparison to MGs from larvae fed on susceptible Tx601 maize that does not express Mir1-CP. Expression of genes encoding proteins involved in PM production is upregulated in MGs from fall armyworm fed on Mp708. Also, several digestive enzymes (endopeptidases, aminopeptidases, lipases, amylase) were more highly expressed in MGs from larvae fed on Mp708 than MGs from larvae fed on Tx601. Impaired growth of larvae fed on Mp708 probably results from metabolic costs associated with higher production of PM constituents and digestive enzymes in a compensatory attempt to maintain MG function. Beginning as neonates, fall armyworm larvae used in the mRNA-seq experiment were reared on yellow-green midwhorl foliage from resistant Mp708 maize or susceptible Tx601 maize. Old foliage and frass were removed every other day and replaced with fresh foliage. Larvae were reared in an environmental chamber at 27M-BM-0C, 14:10 (light:dark) photoperiod, and 70% relative humidity. Midguts were dissected from larvae 2 d after molting to the last instar with masses between 300 and 400 mg. Dissections were done with cold anesthetized larvae submerged in Bombyx saline. After removing Malpighian tubules, foregut anterior to the stomodial valve, hindgut and food bolus, the MG was transferred from the body cavity, rinsed well with cold saline, and preserved in RNAlaterM-BM-.. Equal amounts (3 M-BM-5g) of total RNA from an individual MG were randomly pooled into three replicates per treatment (i.e., Mp708 or Tx601) such that each treatment replicate derived from 12-13 MGs. Each pool of total RNA was separately enriched for poly(A+) RNA and submitted to the Penn State Genomics Core Facility (University Park, PA) where barcoded cDNA libraries were prepared and equimolar quantities of each library sequenced on the SOLiD 3 Plus System. Sequence reads were filtered to accept reads whose median score threshold was M-bM-^IM-%12, contained M-bM-^IM-%25 bases and contained one or more bases with a quality score M-bM-^IM-%14. The 138911 Sanger ESTs in SPODOBASE (http://bioweb.ensam.inra.fr/spodobase) were assembled into a reference transcriptome using SeqMan Pro version 8.0.2. Filtered reads from each library representing a replicate within a maize inbred treatment were mapped separately to the reference transcriptome using the Bowtie-like algorithm in NextGENeM-BM-. with the requirement that 85% of 12 or more nucleotides comprising a read must match the reference. A read was allowed to map only once (i.e., no ambiguous mapping). The number of mapped reads per contig (i.e., gene model) in each treatment replicate-library were summed by NextGENeM-BM-. as read counts per gene and subsequently used in differential expression analyses.
Project description:Here we report 16s rRNA data in gut microbiota of hepatocellular carcinoma (HCC) patients with HBV induced HCC (HBVC) and non-HBV induced HCC (NHBVC) compared with healthy volunteers. A total of 2047 operational taxonomic units (OTUs) were identified in the sequence data. Our data shows that the NHBVC patients harbor lower anti-inflammatory bacteria and more pro-inflammatory bacteria, while the HBVC patients harbor more anti-inflammatory bacteria.
Project description:Opioids such as morphine have many beneficial properties as analgesics, however, opioids may induce multiple adverse gastrointestinal symptoms. We have recently demonstrated that morphine treatment results in significant disruption in gut barrier function leading to increased translocation of gut commensal bacteria. However, it is unclear how opioids modulate the gut homeostasis. By using a mouse model of morphine treatment, we studied effects of morphine treatment on gut microbiome. We characterized phylogenetic profiles of gut microbes, and found a significant shift in the gut microbiome and increase of pathogenic bacteria following morphine treatment when compared to placebo. In the present study, wild type mice (C57BL/6J) were implanted with placebo, morphine pellets subcutaneously. Fecal matter were taken for bacterial 16s rDNA sequencing analysis at day 3 post treatment. A scatter plot based on an unweighted UniFrac distance matrics obtained from the sequences at OTU level with 97% similarity showed a distinct clustering of the community composition between the morphine and placebo treated groups. By using the chao1 index to evaluate alpha diversity (that is diversity within a group) and using unweighted UniFrac distance to evaluate beta diversity (that is diversity between groups, comparing microbial community based on compositional structures), we found that morphine treatment results in a significant decrease in alpha diversity and shift in fecal microbiome at day 3 post treatment compared to placebo treatment. Taxonomical analysis showed that morphine treatment results in a significant increase of potential pathogenic bacteria. Our study shed light on effects of morphine on the gut microbiome, and its role in the gut homeostasis.
Project description:We used 16S V3/V4 region amplification to evaluate the composition of bacteria species in mouse fecal pellets. Fecel pellets were collected from young-adult (12 weeks old) wild type C57Bl/6 mice and aged (72 weeks old) wild type C57Bl/6 mice after 21 days of vehicle or antibiotics treatment (to induce gut microbiota depletion). In one sequencing round, we sequenced a total of 12 different fecal samples (3 young control, 3 aged control, 3 young depleted gut microbiota (ABX) and 3 aged depleted gut microbiota (ABX)). Amplicons were indexed using the Nextera XT Index Kit and pooled into a library for Illumina sequencing.
Project description:Genome scale metabolic model of Drosophila gut microbe Acetobacter fabarum
Abstract -
An important goal for many nutrition-based microbiome studies is to identify the metabolic function of microbes in complex microbial communities and their impact on host physiology. This research can be confounded by poorly understood effects of community composition and host diet on the metabolic traits of individual taxa. Here, we investigated these multiway interactions by constructing and analyzing metabolic models comprising every combination of five bacterial members of the Drosophila gut microbiome (from single taxa to the five-member community of Acetobacter and Lactobacillus species) under three nutrient regimes. We show that the metabolic function of Drosophila gut bacteria is dynamic, influenced by community composition, and responsive to dietary modulation. Furthermore, we show that ecological interactions such as competition and mutualism identified from the growth patterns of gut bacteria are underlain by a diversity of metabolic interactions, and show that the bacteria tend to compete for amino acids and B vitamins more frequently than for carbon sources. Our results reveal that, in addition to fermentation products such as acetate, intermediates of the tricarboxylic acid (TCA) cycle, including 2-oxoglutarate and succinate, are produced at high flux and cross-fed between bacterial taxa, suggesting important roles for TCA cycle intermediates in modulating Drosophila gut microbe interactions and the potential to influence host traits. These metabolic models provide specific predictions of the patterns of ecological and metabolic interactions among gut bacteria under different nutrient regimes, with potentially important consequences for overall community metabolic function and nutritional interactions with the host.IMPORTANCE Drosophila is an important model for microbiome research partly because of the low complexity of its mostly culturable gut microbiota. Our current understanding of how Drosophila interacts with its gut microbes and how these interactions influence host traits derives almost entirely from empirical studies that focus on individual microbial taxa or classes of metabolites. These studies have failed to capture fully the complexity of metabolic interactions that occur between host and microbe. To overcome this limitation, we reconstructed and analyzed 31 metabolic models for every combination of the five principal bacterial taxa in the gut microbiome of Drosophila This revealed that metabolic interactions between Drosophila gut bacterial taxa are highly dynamic and influenced by cooccurring bacteria and nutrient availability. Our results generate testable hypotheses about among-microbe ecological interactions in the Drosophila gut and the diversity of metabolites available to influence host traits.