Project description:The hemibiotrophic fungal pathogen Colletotrichum graminicola is the causal agent of anthracnose disease on maize stalks and leaves. After the formation of appressoria the host cell wall is penetrated by the conversion of appressorial turgor pressure into forceful ejection of a penetration peg. Subsequently, C. graminicola establishes biotrophic hyphae in the penetrated epidermis cell at around 36 hours post inoculation (hpi) until a switch of hyphal morphology and lifestyle takes place during the colonization of neighboring host cells at around 72 hpi. During the ensuing necrotrophic growth, dark necrotic lesions are formed that are visible as anthracnose symptoms. We used microarrays to detail the global programme of gene expression during the infection process of Colletotrichum graminicola in its host plant to get insight into the defense response of this compatible interaction and into the metabolic reprogramming needed to supply the fungus with nutrients. In three independent experiments, maize plants were infected with conidia of the Colletotrichum graminicola strain CgM2 by spray inoculation of leaves. Samples from infected leaves were taken at 36 and 96 hours post infection, corresponding to initial biotrophic and necrotrophic phase, respectively. Samples from uninfected control plants were taken at the same time points.
Project description:The hemibiotrophic fungal pathogen Colletotrichum graminicola is the causal agent of anthracnose disease on maize stalks and leaves. After the formation of appressoria the host cell wall is penetrated by the conversion of appressorial turgor pressure into forceful ejection of a penetration peg. Subsequently, C. graminicola establishes biotrophic hyphae in the penetrated epidermis cell at around 36 hours post inoculation (hpi) until a switch of hyphal morphology and lifestyle takes place during the colonization of neighboring host cells at around 72 hpi. During the ensuing necrotrophic growth, dark necrotic lesions are formed that are visible as anthracnose symptoms. We used microarrays to detail the global programme of gene expression during the infection process of Colletotrichum graminicola in its host plant to get insight into the defense response of this compatible interaction and into the metabolic reprogramming needed to supply the fungus with nutrients.
Project description:Transcriptome of 3 developmental stages of Colletotrichum graminicola during infection of Zea mays leaf sheaths 3 biological replicates per stage. The three stages are: pre-penetration appressoria (PA), early biotrophic phase (BP), and the switch from biotrophy to necrotrophy (NP). Each biological replicate of the first stage, the pre-penetration appressoria, was sequenced to a 2-fold greater depth due to its lower representation in the samples.
Project description:Anthracnose caused by the ascomycete Colletotrichum graminicola is one of the most severe fungal diseases of Zea maize. Cultivars with different levels of resistance have been described. However, which genes contribute to cultivar-specific constitutive and/or induced defense in this economically important pathosystem is still elusive. Transcriptome analyses of infected maize leaves of varieties Golden Jubilee (GJ) and B73 by RNA-Seq was performed for the penetration, biotrophic and necrotrophic phases.
Project description:In order to characterize defense responses not only cytologically, but also on the transcript level, genome-wide sequencing of mRNA isolated from non-infected control leaves and from leaves inoculated either with the WT or with GLS1 overexpressing strains was performed, using Illumina Next Generation Sequencing Technology. In order to identify transcripts specifically induced in leaves infected by β-1,3-glucan-exposing strains, transcript patterns of leaves inoculated with GLS1 overexpressing PtrpC:GLS1 strains were compared with those of the WT. In PtrpC:GLS1-inoculated leaves, a total of 2179 genes were more than 2.5-fold increased, with many genes known as genes typically up-regulated in PAMP-triggered defense responses. These genes include genes encoding PR proteins enzymes involved in cell wall re-inforcemen, and terpene synthases possibly involved in phytoalexin synthesis. Furthermore, increased transcript abundance of genes encoding serine-threonine receptor-like kinases calmodulin, as well as zinc-finger and WRKY transcription factors have been identified. Other up-regulated genes encode proteins involved in protein degradation, i.e. proteases, ubiquitin ligases, as well as enzymes involved in synthesis of auxin or cytokinin phytohormones. In comparison, 2164 genes were more than 2.5-fold down-regulated in maize leaves infected by PtrpC:GLS1 strains, as compared to WT-infected leaves. Several of the encoded proteins are known susceptibility factors. Forty-six down-regulated genes code for proteins containing iron or manganese, or are involved in uptake of these ions, suggesting major re-arrangement of the redox-status in maize leaves after β-glucan perception. Examination of plant defense responses in maize plants inoculated with 2 different Colletotrichum graminicola strains.
Project description:Colletotrichum graminicola is a hemibiotrophic fungal pathogen that causes maize anthracnose disease. It progresses through three recognizable phases of pathogenic development *in planta*: melanized appressoria that form on the host surface prior to penetration; biotrophy, characterized by colonization of living host cells; and necrotrophy, characterized by host cell death and symptom development. An improved filtering algorithm and a Mixed Effects Generalized Linear Model (GLM) were developed and applied to an existing Illumina transcriptome dataset, substantially increasing the statistical power of the analysis of *C. graminicola* gene expression during infection and colonization. Additionally, the *in planta* transcriptome of the wild-type was compared with that of a mutant strain impaired in the establishment of biotrophy, allowing detailed dissection of events occurring specifically during penetration, and during early versus late biotrophy. Results indicated that there is a continuum of activities that occur during colonization of maize by *C. graminicola*, and that boundaries drawn between the three recognizable phases are artificial. More than 2000 fungal genes were differentially transcribed in waves during appressorial maturation, penetration, and colonization. Secreted proteins and membrane receptors were over-represented among the differentially expressed genes, suggesting that the fungus engages in an intimate and dynamic conversation with the host, beginning prior to penetration. This communication process is likely to involve reception of plant signals that trigger subsequent developmental progress in the fungus, as well as the production of signals that induce responses in the host. Later phases of biotrophy were more similar to necrotrophy, with increased production of secreted proteases, inducers of plant cell death, hydrolases, and membrane bound transporters for the uptake and egress of potential toxins, signals, and nutrients. The differentially expressed genes could be used as landmarks to more accurately identify developmental progress in compatible versus incompatible interactions involving genetic variants of both host and pathogen.