Identification of the transcriptional profile of the hyphal tip of Neurospora crassa
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ABSTRACT:
ORGANISM(S): Neurospora crassa
SUBMITTER: Charles Hall
PROVIDER: E-GEOD-50205 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:A colony of fungus is comprised of long, branching filamentous cells called hyphae. Genetic mechanisms underlying the development of hyphae are poorly understood. We sectioned hyphae of a model fungus, Neurospora crassa (pink bread mold), into six parts depending on the age of the cells; 1 hour, 3 hour, 9 hour, 15 hour, 21 hour and 27 hour old, respectively. This data submission reflects an RNAseq analysis of mRNA extracted from the 1 hour time-point.
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Project description:To determine the genes directly and indirectly under the control of the Grainy-head homolog (GHH) transcription factor in Neurospora crassa Three different sample types (Aerial Hyphae & Conidia; Mycelia; or Whole Colonies) of both wild-type (FGSC #2489) and grainy-head homolog (FGSC #13563) strains of Neurospora crassa were subjected to transcriptome analyses to determine the genes differentially expressed in the ghh background compared to wild type.
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Project description:Neurospora tetrasperma is a pseudohomothallic filamentous ascomycete with a large (~ 7 Mbp) region of suppressed recombination surrounding its mating-type (mat) locus. The suppressed recombination has lead to sequence divergence between the two mating-type chromosomes of wild-type heterokaryotic strains, while the remaining genome is largely homoallelic. In this study, we use microarray technology to manifest expression divergence linked to mating type in N. tetrasperma. N. tetrasperma and N. crassa, were grown on agar regimes inducing sexual growth (Synthetic Crossing medium) and vegetative growth (Vogel's Medium), respectively. [SC]: Neurospora tetrasperma mat-A FGSC#1270; mat-a FGSC#1271; Mat-A FGSC#9033; mat-a FGSC#9034; N. crassa mat-A FGSC#2489 and mat-a FGSC 4200: Synthetic Crossing medium was used as a nutrient regime before sampling and processing [Veg]: Neurospora tetrasperma mat-A FGSC#1270; mat-a FGSC#1271; Mat-A FGSC#9033; mat-a FGSC#9034; N. crassa mat-A FGSC#2489 and mat-a FGSC 4200: Vogel's Medium (Vegetative Medium) was used as a nutrient regime before sampling and processing
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Project description:Light represents an important environmental cue, which exerts considerable influence on the metabolism of fungi. Studies with the biotechnological fungal workhorse Trichoderma reesei (Hypocrea jecorina) have revealed an interconnection between transcriptional regulation of cellulolytic enyzmes and the light response. The filamentous fungus, Neurospora crassa, has been used as a model organism to study light and circadian rhythm biology. We therefore investigated whether light also regulates transcriptional regulation of cellulolytic enzymes in N. crassa. We show that the N. crassa photoreceptor genes wc-1, wc-2 and vvd are involved in regulation of cellulase gene expression, indicating that this phenomenon is conserved among filamentous fungi. Genome wide analysis of photoreceptor mutants and evaluation of results by analysis of mutant strains identified several candidate genes likely to play a role in light modulated cellulase gene expression. Three deletion strains (delta-wc-1 (FGSC 11712), delta-wc-2 (FGSC 11124) and delta-vvd (FGSC 11556)) and the wild type strain (FGSC 2489) at two different timepoints (28h or 40h) were analyzed. Cy3 and Cy5 dye swaps were performed.
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Project description:Multi-targeting priming (MTP) for genome-wide gene expression assays provides selective targeting of multiple sequences and counter-selection against undesirable sequences. We demonstrated superior performance of two MTPs compared to oligo-dT microarray profling and RNA tag sequencing the response of Saccharomyces cerevisiae to nitrogen deficiency and profiling Neurospora crassa early sexual development. Priming with MTPs in addition to oligo-dT resulted in higher sensitivity, a greater number of well-measured genes, more genes significantly differentially expressed, and a greater power to detect meager differences. Neurospora crassa mat A FGSC#2489 Three developmental stages and two different primers used for reverse transcription: mycelium oligo(dT) M1 protoperithecia oligo(dT) PP1 perithecia oligo(dT) PT1 mycelium oligo(dT)+ Multi-Targeted Primer [MTP] (M2) protoperithecia oligo(dT)+ MTP (PP2) perithecia oligo(dT)+ MTP (PT2)
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Project description:Understanding how genomes encode complex cellular and organismal behaviors is an outstanding challenge of modern genetics. Unlike classical screening methods, analysis of the genetic variation that occurs naturally in wild populations can enable rapid, genome-scale trait mapping with a medium-throughput experimental design. Here we describe the results of the first genome-wide association analysis (GWAS) of a microbial eukaryote, using wild isolates of the filamentous fungus Neurospora crassa. We transcriptionally profiled each of 112 individuals collected in Louisiana and used RNA-seq data to genotype each strain at thousands of genetic loci genome-wide. We used these genotypes in a mapping analysis of microbial communication. In N. crassa, germinated asexual spores (germlings) sense the presence of other germlings, grow toward them in a coordinated fashion, and fuse. We evaluated germlings of each strain for their ability to chemically sense, chemotropically seek, and undergo cell fusion. GWAS identified one gene, NCU04379 (cse-1, encoding a homolog of neuronal calcium sensor), at which inheritance was strongly associated with the efficiency of germling communication. Deletion of cse-1 significantly lowered germling communication and fusion, and two genes encoding predicted binding partners of CSE-1 were also required for the communication trait. Additionally, mining our association results for signaling and secretion genes with a potential role in germling communication, we validated six more novel determinants of germling communication, including a secreted protease and two other genes whose deletion conferred a novel phenotype of increased communication and multi-germling fusion. These results establish protein secretion as a linchpin of germling communication in N. crassa and shed light on the regulation of communication molecules in this fungus. Our study demonstrates the power of population-genetic analyses for the rapid identification of genes contributing to complex traits in microbial species. 112 mycelial mRNA profiles of 16 hour wild type (WT) Neurospora crassa strains were generated by deep sequencing, using Illumina GAIIx.
2013-03-28 | E-GEOD-45406 | biostudies-arrayexpress
Project description:It is widely assumed that sex-determination loci have little impact upon dominant stage in the life history of filamentous fungi, asexual development. However, this hypothesis has not been thoroughly investigated. To investigate differences between mating types during asexual development, total RNA was sampled from otherwise largely isogenic Mat-A and Mat-a N. crassa strains at early, middle, and late clonal stages of development under a condition of constant light. Mating-type loci, pheromone precursor and receptor genes were assayed with real-time PCR. Mating loci were increasingly expressed during asexual development, and expression of pheromone precursors ccg-4 and mfa-1 and receptors pre-1 and pre-2 were detected in both mating types in all development stages. A circuit of microarray hybridizations of cDNA samples from all developmental stages was also performed. Gene expression for both mating types throughout vegetative development were characterized for developmental markers such as transcription factors, for genes related to conidiation, internal clock, cell division cycle, heat shock, and for light responsive genes. We observed significant differences in overall gene expression between the mating types across clonal development, especially at late development stages. The Mat-A genotype showed a higher expression level than Mat-a for many genes, and demonstrated greater transcriptional regulatory activity. Significant up-regulation of expression was observed for many late light responsive genes at late asexual development stages in both mating types. Further investigation of the impact of light and the roles of light response genes in asexual development in this fungus are warranted. Neurospora crassa mat A FGSC#2489 and mat-a FGSC 4200: Three developmental stages, 36hr, 60hr, 96 hr were sampled and compared
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