Stachybotrys elegans is a mycoparasite of the soilborne plant pathogen fungus Rhizoctonia solani. The mycoparasitic activity of S. elegans is correlated with the production of cell wall-degrading enzymes such as chitinases. This study details the cloning and characterization of the cDNA, sechi44, that encodes an extracellular endochitinase. The expression regulation of sechi44 was altered when S. elegans was in interaction with its host, R. solani, and also when the mycoparasite was grown on minimal media amended with different carbon and nitrogen sources. Direct contact with R. solani significantly upregulated sechi44 expression which followed a cyclical pattern suggesting that this gene has a role not only in mycoparasitism, but also in linear growth of the mycoparasite. The addition of high concentrations of glucose and ammonium triggered a decrease of sechi44 expression suggesting that sechi44 is subject to glucose and ammonium repression. In a separate study, several genes (1016 clones) whose transcription was substantially up-regulated during the mycoparasitic interaction were identified using SSH and microarray analysis. Twenty-five percent (261 clones) of these were sequenced and assigned to putative functions. Among them, 15 expressed sequence tags (ESTs) were identified in R. solani whose functions were related to defense while the majority of ESTs were identified in S. elegans and assigned functions related to toxin metabolism, pathogenic process, stress response., multidrug resistance, apoptosis, transport, ATP synthesis, replication, transcription and DNA repair, translation, transduction, protein degradation, and ribosomal protein. The overexpression of 13 selected genes of S. elegans was validated and confirmed using quantitative reverse transcription polymerase chain reaction (QRT-PCR). The temporal gene expression of nine genes was monitored when the mycoparasite was grown on R. solani (host) and Sclerotinia sclerotiorum (non-host) mycelia and sclerotia. Some genes such as seglu, selec, and se151 were completely inhibited by the presence of non-host hyphae suggesting that these genes play an important role during mycoparasitism. Also, the absence of these corresponding transcripts suggests that the non-host produces transcription inhibitors. As expected, gene expression of cytochrome P450 was highly up-regulated early after germination of S. elegans conidia. This is in agreement with our finding in the EST data mining study, in which a role in toxin production was assigned to cytochrome P450.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.102815 |
| Date | January 2006 |
| Creators | Morissette, Danielle. |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Doctor of Philosophy (Department of Plant Science.) |
| Rights | © Danielle Morissette, 2006 |
| Relation | alephsysno: 002601702, proquestno: AAINR32220, Theses scanned by UMI/ProQuest. |
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