Identification by RNA fingerprinting of genes differentially expressed during the development of the basidiomycete Lentinula edodes

Abstract

As part of an ongoing project to understand the molecular mechanisms of fruit body development in Lentinula edodes (Shiitake mushroom), RNA fingerprinting by arbitrarily primed PCR (RAP-PCR) was used to identify differentially expressed genes in RNA populations from four stages of L. edodes development vegetative mycelium, primordium, young fruit body and mature fruit body. From 30 RNA fingerprints, we cloned and sequenced 33 RAP fragments after their differential expression patterns had been verified by reverse Northern dot-blot hybridization. Thirteen RAP fragments show high sequence similarity to known gene products which are involved in (1) transport across the plasma membrane (drug efflux pump and sugar transporter); (2) cell cycle control (cyclin B); (3) signal transduction and transcriptional regulation (mitogen-activated protein kinase, Cdc39/Not1, PriA, Jun-D); (4) intracellular molecule trafficking (ubiquitin, plasma membrane proton ATPase, and alpha-adaptin); (5) mitochondrial biogenesis (mitochondrial processing peptidase beta-subunit, mitochondrial glycerol-3-phosphate dehydrogenase); and (6) intermediary metabolism (fructose 1,6 bisphosphatase). The transcript levels for plasma membrane proton ATPase and alpha-adaptin remained constant, whereas the other eleven genes were differentially expressed during L. edodes. development. The expression profiles of the genes suggest that transport across the plasma membrane is important in the mycelial stage. Specific signal transduction and transcriptional controls may play important roles during the initiation of primordia and the formation of young fruiting bodies. When the mushroom matures, expression of genes involved in metabolic pathways becomes prominent. The isolation of these genes indicates their involvement in homobasidiomycete development and suggests new directions for molecular studies on mechanisms of mushroom development.

Isolation and Characterization of Differentially Expressed Genes in the Mycelium and Fruit Body of Tuber borchii

“…Forty-one transcripts showed striking up-regulation at either of the developmental stages, suggesting that the corresponding proteins are directly implicated in the morphogenesis and functioning of fruit bodies during spore maturation. In Lentinula edodes (19), A. bisporus (8), and A. aegerita (32), similar numbers of fruit body-regulated genes were identified. Sixteen genes, mostly coding for unknown proteins, had lower transcript concentrations and probably represented vegetative mycelium-specific genes.…”

Chemical stress-responsive genes from the lignin-degrading fungus Phanerochaete chrysosporium exposed to dibenzo-p-dioxin

“…Thus, we screened dioxin-mediated stress response genes to better understand the cellular response of P. chrysosporium utilizing a DDRT-PCR technique. Recently, a successful application of this technique to study fungal cellular response was reported [10,12,13].…”

Characterization of a Pleurotus ostreatus fruiting body‐specific hydrophobin gene, Po.hyd

“…About 20 µg of each sample was separated on a 1.0% denaturing agarose-formaldehyde gel, transferred onto a Hybond-N + nylon filter (Amersham Biosciences) and then fixed by UV cross-linking (Stratagene) (Leung et al 2000). The cDNA probe corresponding to Po.hyd was prepared using the random priming procedure (Megaprime DNA labeling System, Amersham Biosciences).…”