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Analysis of transcript abundance during appressorium formation in the rice blast fungus Magnaporthe oryzae using HT-SuperSAGE.

ABSTRACT: The rice-blast fungus Magnaporthe oryzae is disseminated using a three-celled spore or conidium. Upon landing on the surface of a rice leaf, the conidium germinates and forms a specialised structure for entry into the host plant - the appressorium. In this study we have followed gene expression thoughout germination of the conidium and formation of the appressorium to identify genes that may be important for appressorium function. We have also compared gene expression in the wild-type germinating conidium at four hours to a mutant strain deleted for the MAP kinase pmk1. The pmk1 mutant is unable to form appressoria.

ORGANISM(S): Pyricularia oryzae

PROVIDER: GSE30069 | GEO | 2012/02/13

SECONDARY ACCESSION(S): PRJNA144005

REPOSITORIES: GEO

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Analysis of transcript abundance during appressorium formation in the rice blast fungus Magnaporthe oryzae using HT-SuperSAGE.

Project description:The rice-blast fungus Magnaporthe oryzae is disseminated using a three-celled spore or conidium. Upon landing on the surface of a rice leaf, the conidium germinates and forms a specialised structure for entry into the host plant - the appressorium. In this study we have followed gene expression thoughout germination of the conidium and formation of the appressorium to identify genes that may be important for appressorium function. We have also compared gene expression in the wild-type germinating conidium at four hours to a mutant strain deleted for the MAP kinase pmk1. The pmk1 mutant is unable to form appressoria. Samples were taken from five time-points in wild-type germinating conidia and one time-point in the pmk1 mutant. Two replicates were taken at each time-point.

2012-02-13 | E-GEOD-30069 | biostudies-arrayexpress

Phosphoproteome Analysis of Magnaporthe oryzae During Appressorium Development

Project description:The rice pathogen, Magnaporthe oryzae, undergoes a complex developmental process leading to formation of an appressorium prior to plant infection. In an effort to better understand phosphoregulation during appressorium development, a mass spectrometry-based study was undertaken. Phosphosites were identified from phosphoproteins from mycelia, conidia, germlings, and appressoria of the wild type and a ckpA mutant. The cyclic-AMP dependent protein kinase A (CPKA) is required for initial perception of the leaf surface, development of functional appressoria, and timely mobilization of storage reserves in conidia, including glycogen and lipid bodies.

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Hierarchical transcriptional network controls appressorium-mediated plant infection by the rice blast fungus Magnaporthe oryzae

Project description:We report that the homeobox trasncription factor called HOX7 controls hyphae-associated genes, autophagy and cell cycle related genes necessary for appressorium development in the rice blast fungus Magnaporthe oryzae. We also report that the ste transcription factor MST12 regulates gene functions involved in septin reorientation.

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A hierarchical transcriptional network controls appressorium-1 mediated plant 2 infection by the rice blast fungus Magnaporthe oryzae

Project description:Rice blast is a pervasive and devastating disease that threatens rice production across the world. In spite of its importance to global food security, however, the underlying biology of plant infection by the blast fungus Magnaporthe oryzae remains poorly understood. In particular, it is unclear how the fungus elaborates a specialised infection cell, the appressorium, in response to surface signals from the rice leaf. Here, we report the identification of a network of temporally co-regulated transcription factors that act downstream of the Pmk1 mitogen-activated protein kinase pathway to regulate gene expression during appressorium-mediated plant infection. We have functionally characterised this network of transcription factors and demonstrated the operation of a hierarchical transcriptional control system. We show that this tiered regulatory mechanism involves Pmk1-dependent phosphorylation of the Hox7 homeobox transcription factor, which represses hyphal-associated gene expression and simultaneously induces major physiological changes required for appressorium development, including cell cycle arrest, autophagic cell death, turgor generation and melanin biosynthesis. Mst12 then regulates gene functions involved in septin-dependent cytoskeletal re-organisation, polarised exocytosis and effector gene expression necessary for plant tissue invasion.4929

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Germination and appressorium formation in Magnaporthe grisea

Project description:This series examines differential gene expression during germination and appressorium formation by the rice blast fungus Magnaporthe grisea. Conidia were germinated on either the hydrophobic (inductive) or hydrophilic (non-inductive) side of GelBond. RNA was extracted from conidia and following incubation for 7 and 12 hours. RNA from two biological replications of each treatment was pooled in equal amounts and labeled with both cy3 and cy5 dyes using the Agilent Technologies Low Input Linear Amplification Kit. Hybridizations were performed on Agilent Technologies whole genome oligo rice blast arrays (product G4137A) using manufacturer prescribed protocols and reagents in an interlaced loop design in which each treatment was paired with every other. This series contains a total of 10 hybridizations; each treatment was used in 4 hybridizations (2 with cy3 and 2 cy5). Spot fluorescence was normalized using Lowess within and between microarray slides in Bioconductor and gene expression profiles analyzed in GeneSpring. Keywords: other

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Magnaporthe appressoria: wt vs. bip1 deletion mutant

Project description:The identification of genes regulated by the transcription factor Bip1 during appressorial development. The rice blast fungus Magnaporthe grisea develops melanized appressoria with high turgor pressure that inject the fungus into epidermal host cells. A screen for M. grisea non-pathogenic mutants recovered mutant M763 with reduced penetration efficiency and infectious growth. Complementation analysis identified a gene inactivated in M763 that encodes a novel basic leucine zipper (bZIP) transcription factor named Bip1 (B-ZIP Involved in Pathogenesis1). Deletion of Bip1 by targeted replacement generated non-pathogenic mutants that are unable to infect rice or barley. These mutants develop melanized appressoria that are unable to penetrate intact or wounded host leaves. Bip1 mRNA was detected in fungal spores, appressoria and infected leaves but not in mycelia, while a BIP1:GFP fusion was detected only in nuclei of mature appressoria. Genome wide transcriptome analysis identified 44 genes down regulated in Δbip1:hph appressoria. Most of these genes (77%) are specifically expressed in appressoria and encode enzymes involved in secondary metabolism (11), secreted proteins and enzymes (18) and G-Protein Coupled Receptors (GPCRs) (5). Promoters of BIP1 down-regulated genes share a similar GCN4/bZIP DNA binding motif (TGACTC). Gel shift assays showed that the motifs from 6 of the 7 promoters tested bind to recombinant BIP1 in vitro, including MGG_08381.5 from the ACE1 locus. Mutation of BIP1 binding sites in the MGG_08381.5 promoter significantly reduced its appressorium specific expression. Molecular and biochemical studies confirmed that BIP1 controls the expression of a distinct set of M. grisea genes in the appressorium, revealing a novel regulatory network involved in early stages of fungal infection that activates a diverse gene expression program resulting in a “shock and awe” impact on the plant host.

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Germination and appressorium formation in Magnaporthe grisea

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Project description:This series examines differential gene expression during appressorium formation induced by exogeous cyclic AMP in the rice blast fungus Magnaporthe grisea. RNA was extracted from spores germinated on hydrophilic (non-inductive) side of GelBond as well as from spores elaborating appressia on hydrophilic surfaces in the presence of cAMP after 9 hrs incubation. RNA from two biological replications of each treatment was pooled in equal amounts and labeled with both cy3 and cy5 dyes using the Agilent Technologies Low Input Linear Amplification Kit. Hybridizations were performed on Agilent Technologies whole genome oligo rice blast arrays (product G4137B) using manufacturer prescribed protocols and reagents in an reference design in which each treatment was paired with every other. This series contains a total of 4 hybridizations; each treatment was used in 4 hybridizations (2 with Cy3 and 2 Cy5). Spot fluorescence was normalized using Lowess within and between microarray slides in Bioconductor. Keywords: Cell development

2008-01-15 | GSE10173 | GEO

Appressorium induction by cyclic AMP in Magnaporthe grisea

2010-05-26 | E-GEOD-10173 | biostudies-arrayexpress

Differential gene expression of Appresorium vs Vegetative mycelium of Magnaporthe using oligoarray

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