Project description:Regulated host cell death is part of a plant defense strategy against pathogens but it is also involved in accommodating certain beneficial root microbes. We have identified extracellular metabolites and intracellular metabolic signals that contribute to beneficial root fungal endophyte colonization, and uncovered a conserved cell death mechanism likely co-opted for establishing plant-endophyte symbiosis.
Project description:We addressed the question how the interaction between the beneficial root endophyte Serendipita vermifera (Sv) and the pathogen Bipolaris sorokiniana (Bs) affects fungal behavior and determines barley host responses using a gnotobiotic natural soil-based split-root system for phenotypic and transcriptional analyses.
Project description:Broad-host root endophytes establish long-term interactions with a large variety of plants, thereby playing a significant role in natural and managed ecosystems and in evolution of land plants. To exploit plants as living substrates and to establish a compatible interaction with morphologically and biochemically extremely different hosts, endophytes must respond and adapt to different plant signals and host metabolic states. Here we identified host-adapted colonization strategies and host-specific effector candidates of the mutualistic root endophyte Piriformospora indica by a global investigation of fungal transcriptional responses to barley and Arabidopsis at different symbiotic stages. Additionally we examined the role played by nitrogen in these two diverse associations. Cytological studies and colonization analyses of a barley mutant and fungal RNAi strains show that distinct physiological and metabolic signals regulate host-specific lifestyle in P. indica. This is the foundation for exploring how distinct fungal and host symbiosis determinants modulate biotrophy in one host and saprotrophy in another host and, ultimately, gives hints into the mechanisms underlying host adaptation in root symbioses.
Project description:Broad-host root endophytes establish long-term interactions with a large variety of plants, thereby playing a significant role in natural and managed ecosystems and in evolution of land plants. To exploit plants as living substrates and to establish a compatible interaction with morphologically and biochemically extremely different hosts, endophytes must respond and adapt to different plant signals and host metabolic states. Here we identified host-adapted colonization strategies and host-specific effector candidates of the mutualistic root endophyte Piriformospora indica by a global investigation of fungal transcriptional responses to barley and Arabidopsis at different symbiotic stages. Additionally we examined the role played by nitrogen in these two diverse associations. Cytological studies and colonization analyses of a barley mutant and fungal RNAi strains show that distinct physiological and metabolic signals regulate host-specific lifestyle in P. indica. This is the foundation for exploring how distinct fungal and host symbiosis determinants modulate biotrophy in one host and saprotrophy in another host and, ultimately, gives hints into the mechanisms underlying host adaptation in root symbioses. Arabidopsis and barley roots were inoculated with Piriformospora indica and grown for 14 days. Additionally P. indica was grown on 1/10 PNM medium alone. Samples were taken 3 and 14 dpi (Arabidopsis), 14 dpi (barley) and 3dpi (1/10 PNM). Each experiment was performed in three independent biological repetitions. Piriformospora indica gene expression examined only.
Project description:The root-colonizing fungal endophyte Serendipita indica, formerly known as Piriformospora indica, is well known to promote plant biomass production and stress tolerance of its host plants. Co-cultivation of Arabidopsis thaliana seedlings with the fungus triggers a substantial induction of the growth of the root system. However, the molecular mechanisms by which the fungus promotes plant growth over an extended period of time is still unclear. We here report the comparative analysis of the effect of a mock- and S. indica-infection on wild-type Arabidopsis plants (Col-0) after 2 and 10 days of co-cultivation. Our data provide evidence for the induction of a number of genes that are consistingly induced during the plant–fungus interaction.
Project description:Inoculation of endophyte-free (E-) Theobroma cacao leaves with Colletotrichum tropicale (E+), the dominant foliar fungal endophyte in healthy T. cacao, induced significant changes in the expression of hundreds of host genes. Further, E+ leaves exhibit enhanced pathogen resistance, increased lignin and cellulose content, reduced maximum rates of photosynthesis (Amax), and enrichment of nitrogen-15 and carbon-13 isotopes that all correspond to the changes in expression of specific functional genes in related pathways. Moreover, a cacao gene highly up-regulated in E+ leaves increases pathogen resistance apart from any direct endophyte effects. Thus, benefits of increased pathogen resistance in E+ plants are partially due to enhanced induction of intrinsic host defense pathways, and potential costs include reduced photosynthetic capacity and endophyte metabolism of host tissues. Similar effects are likely to be properties of most plant-endophyte interactions, suggesting general relevance to the design and interpretation of studies of genetic and phenotypic expression in plants. The objective of this experiment was to identify Theobroma cacao genes that are differentially expressed between leaves inoculated with fungal endophyte Colletotrichum tropicale (E+ leaves) and control un-inoculated leaves (E- leaves) 3 days post endophyte inoculation. The experiment was conducted in a Percival growth chamber (model I35LL, 115 volts, 1/4 Hp, series: 8503122.16, Percival Scientific, Inc., Perry IA) with 12/12 h light/dark photoperiod and temperatures of 30M-BM-:C and 26M-BM-:C respectively. Inoculation was done by aspersion of endophyte spores (2X10^6 spore/ml) to a group of T. cacao seedlings and a second group of seedlings were maintained as control un-inoculated (E- leaves). Then three biological replicates (each one consisting of one leaf from different plants) per treatment E+ and four leaves per treatment E- leaves) were collected and processed for a two color oligo microarray analysis.
Project description:As plant roots are built of concentric cell-layers, it is anticipated that these respond to microbial infection by employing specific, genetically defined programs. Due to cell-type specific expression of tagged ribosomes, ribosome-bound mRNA was isolated to obtain cell-layer translatomes. After inoculation with the vascular pathogen Verticillium longisporum, the pathogenic oomycete Phytophthora parasitica and the mutualistic endophyte Serendipita indica, we determined root responses on a cell-layer resolution. These data reflect the fundamentally different colonization strategies of the microbes. Mining the data-set, we identified a Verticillium specific suppression of the endodermal barrier restricting fungal progression, localized biosynthesis of antimicrobial compounds, and observed a pathogen-specific arrest of root meristems correlated with ethylene biosynthesis. These examples highlight the power of this approach in generating testable hypotheses to gain insights into Root-microbe-interactions.
Project description:Inoculation of endophyte-free (E-) Theobroma cacao leaves with Colletotrichum tropicale (E+), the dominant foliar fungal endophyte in healthy T. cacao, induced significant changes in the expression of hundreds of host genes. Further, E+ leaves exhibit enhanced pathogen resistance, increased lignin and cellulose content, reduced maximum rates of photosynthesis (Amax), and enrichment of nitrogen-15 and carbon-13 isotopes that all correspond to the changes in expression of specific functional genes in related pathways. Moreover, a cacao gene highly up-regulated in E+ leaves increases pathogen resistance apart from any direct endophyte effects. Thus, benefits of increased pathogen resistance in E+ plants are partially due to enhanced induction of intrinsic host defense pathways, and potential costs include reduced photosynthetic capacity and endophyte metabolism of host tissues. Similar effects are likely to be properties of most plant-endophyte interactions, suggesting general relevance to the design and interpretation of studies of genetic and phenotypic expression in plants. The objective of this experiment was to identify Theobroma cacao genes that are differentially expressed between leaves inoculated with fungal endophyte Colletotrichum tropicale (E+ leaves) and control un-inoculated leaves (E- leaves) 14 days post last endophyte inoculation. The experiment was conducted in a Percival growth chambers (model I35LL, 115 volts, 1/4 Hp, series: 8503122.16, Percival Scientific, Inc., Perry IA) with 12/12 h light/dark photoperiod and temperatures of 30M-BM-:C and 26M-BM-:C respectively. A total of four endophyte spore inoculations (1X10^6 spore/ml) were made by aspersion to a group of T. cacao seedlings and a second group of seedlings were maintained as un-inoculated. Then six biological replicates per treatment (E+ leaves and six E- leaves) each one belonging from a different seedling were collected and processed for a two color oligo microarray analysis. A total of six arrays were processed, each one hybridized to an inoculated and a control un-inoculated sample in a dye swap design.
Project description:The root-colonizing fungal endophyte Serendipita indica, formerly known as Piriformospora indica, is well known to promote plant biomass production and stress tolerance of its host plants. Moreover, previous studies highlighted an important impact of the fungus on auxin homeostasis during the infection of Arabidopsis thaliana plants. Auxin is a key determinant of plant growth, including the growth of the root system. Auxin overproducing mutants, like for instance YUC9oe (Hentrich et al., 2013 Plant J.), show a pronounced root phenotype that can be restored by the co-cultivation with S. indica. We here report the comparative analysis of the effect of a mock- and S. indica-infection on both wild-type Arabidopsis plants (Col-0) and YUC9 overexpressing mutants. Our data provide evidence for the induction of GRETCHEN HAGEN 3 (GH3) genes that are involved in conjugating active free indole-3-acetic acid with amino acids. The fungus triggered induction GH3s is suggested to be involved in affecting the cellular auxin homeostasis.
Project description:Inoculation of endophyte-free (E-) Theobroma cacao leaves with Colletotrichum tropicale (E+), the dominant foliar fungal endophyte in healthy T. cacao, induced significant changes in the expression of hundreds of host genes. Further, E+ leaves exhibit enhanced pathogen resistance, increased lignin and cellulose content, reduced maximum rates of photosynthesis (Amax), and enrichment of nitrogen-15 and carbon-13 isotopes that all correspond to the changes in expression of specific functional genes in related pathways. Moreover, a cacao gene highly up-regulated in E+ leaves increases pathogen resistance apart from any direct endophyte effects. Thus, benefits of increased pathogen resistance in E+ plants are partially due to enhanced induction of intrinsic host defense pathways, and potential costs include reduced photosynthetic capacity and endophyte metabolism of host tissues. Similar effects are likely to be properties of most plant-endophyte interactions, suggesting general relevance to the design and interpretation of studies of genetic and phenotypic expression in plants.