Cargando…
Combining laser microdissection and RNA-seq to chart the transcriptional landscape of fungal development
BACKGROUND: During sexual development, filamentous ascomycetes form complex, three-dimensional fruiting bodies for the protection and dispersal of sexual spores. Fruiting bodies contain a number of cell types not found in vegetative mycelium, and these morphological differences are thought to be med...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2012
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3472292/ https://www.ncbi.nlm.nih.gov/pubmed/23016559 http://dx.doi.org/10.1186/1471-2164-13-511 |
_version_ | 1782246574328381440 |
---|---|
author | Teichert, Ines Wolff, Gabriele Kück, Ulrich Nowrousian, Minou |
author_facet | Teichert, Ines Wolff, Gabriele Kück, Ulrich Nowrousian, Minou |
author_sort | Teichert, Ines |
collection | PubMed |
description | BACKGROUND: During sexual development, filamentous ascomycetes form complex, three-dimensional fruiting bodies for the protection and dispersal of sexual spores. Fruiting bodies contain a number of cell types not found in vegetative mycelium, and these morphological differences are thought to be mediated by changes in gene expression. However, little is known about the spatial distribution of gene expression in fungal development. Here, we used laser microdissection (LM) and RNA-seq to determine gene expression patterns in young fruiting bodies (protoperithecia) and non-reproductive mycelia of the ascomycete Sordaria macrospora. RESULTS: Quantitative analysis showed major differences in the gene expression patterns between protoperithecia and total mycelium. Among the genes strongly up-regulated in protoperithecia were the pheromone precursor genes ppg1 and ppg2. The up-regulation was confirmed by fluorescence microscopy of egfp expression under the control of ppg1 regulatory sequences. RNA-seq analysis of protoperithecia from the sterile mutant pro1 showed that many genes that are differentially regulated in these structures are under the genetic control of transcription factor PRO1. CONCLUSIONS: We have generated transcriptional profiles of young fungal sexual structures using a combination of LM and RNA-seq. This allowed a high spatial resolution and sensitivity, and yielded a detailed picture of gene expression during development. Our data revealed significant differences in gene expression between protoperithecia and non-reproductive mycelia, and showed that the transcription factor PRO1 is involved in the regulation of many genes expressed specifically in sexual structures. The LM/RNA-seq approach will also be relevant to other eukaryotic systems in which multicellular development is investigated. |
format | Online Article Text |
id | pubmed-3472292 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-34722922012-10-17 Combining laser microdissection and RNA-seq to chart the transcriptional landscape of fungal development Teichert, Ines Wolff, Gabriele Kück, Ulrich Nowrousian, Minou BMC Genomics Research Article BACKGROUND: During sexual development, filamentous ascomycetes form complex, three-dimensional fruiting bodies for the protection and dispersal of sexual spores. Fruiting bodies contain a number of cell types not found in vegetative mycelium, and these morphological differences are thought to be mediated by changes in gene expression. However, little is known about the spatial distribution of gene expression in fungal development. Here, we used laser microdissection (LM) and RNA-seq to determine gene expression patterns in young fruiting bodies (protoperithecia) and non-reproductive mycelia of the ascomycete Sordaria macrospora. RESULTS: Quantitative analysis showed major differences in the gene expression patterns between protoperithecia and total mycelium. Among the genes strongly up-regulated in protoperithecia were the pheromone precursor genes ppg1 and ppg2. The up-regulation was confirmed by fluorescence microscopy of egfp expression under the control of ppg1 regulatory sequences. RNA-seq analysis of protoperithecia from the sterile mutant pro1 showed that many genes that are differentially regulated in these structures are under the genetic control of transcription factor PRO1. CONCLUSIONS: We have generated transcriptional profiles of young fungal sexual structures using a combination of LM and RNA-seq. This allowed a high spatial resolution and sensitivity, and yielded a detailed picture of gene expression during development. Our data revealed significant differences in gene expression between protoperithecia and non-reproductive mycelia, and showed that the transcription factor PRO1 is involved in the regulation of many genes expressed specifically in sexual structures. The LM/RNA-seq approach will also be relevant to other eukaryotic systems in which multicellular development is investigated. BioMed Central 2012-09-27 /pmc/articles/PMC3472292/ /pubmed/23016559 http://dx.doi.org/10.1186/1471-2164-13-511 Text en Copyright ©2012 Teichert et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Teichert, Ines Wolff, Gabriele Kück, Ulrich Nowrousian, Minou Combining laser microdissection and RNA-seq to chart the transcriptional landscape of fungal development |
title | Combining laser microdissection and RNA-seq to chart the transcriptional landscape of fungal development |
title_full | Combining laser microdissection and RNA-seq to chart the transcriptional landscape of fungal development |
title_fullStr | Combining laser microdissection and RNA-seq to chart the transcriptional landscape of fungal development |
title_full_unstemmed | Combining laser microdissection and RNA-seq to chart the transcriptional landscape of fungal development |
title_short | Combining laser microdissection and RNA-seq to chart the transcriptional landscape of fungal development |
title_sort | combining laser microdissection and rna-seq to chart the transcriptional landscape of fungal development |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3472292/ https://www.ncbi.nlm.nih.gov/pubmed/23016559 http://dx.doi.org/10.1186/1471-2164-13-511 |
work_keys_str_mv | AT teichertines combininglasermicrodissectionandrnaseqtochartthetranscriptionallandscapeoffungaldevelopment AT wolffgabriele combininglasermicrodissectionandrnaseqtochartthetranscriptionallandscapeoffungaldevelopment AT kuckulrich combininglasermicrodissectionandrnaseqtochartthetranscriptionallandscapeoffungaldevelopment AT nowrousianminou combininglasermicrodissectionandrnaseqtochartthetranscriptionallandscapeoffungaldevelopment |