Use of 1H NMR to measure intracellular metabolite levels during growth and asexual sporulation in Neurospora crassa.
Eukaryot Cell. 2011 Apr 1;
Authors: Kim JD, Kaiser K, Larive CK, Borkovich KA
Conidiation is an asexual sporulation pathway that is a response to adverse conditions and is the main mode of dispersal utilized by filamentous fungal pathogens for reestablishment in a more favorable environment. Heterotrimeric G proteins (consisting of ?, ? and ? subunits) have been shown to regulate conidiation in diverse fungi. Previous work has demonstrated that all three of the G? subunits in the filamentous fungus Neurospora crassa affect mass accumulation on poor carbon sources and that loss of gna-3 leads to the most dramatic effects on conidiation. In this study, we used (1)H-NMR to profile the metabolome of N. crassa in extracts isolated from vegetative hyphae and conidia from cultures grown under conditions of high or low sucrose. We compared wild-type and ?gna-3 strains to determine whether lack of gna-3 causes a significant difference in global metabolite profile. The results demonstrate that the global metabolome of wild-type hyphae is influenced by carbon availability. The metabolome of the ?gna-3 strain cultured on both high and low sucrose is similar to wild-type grown on high sucrose, suggesting an overall defect in nutrient sensing in the mutant. However, analysis of individual metabolites revealed differences in wild-type and ?gna-3 strains cultured under low and high sucrose.
PMID: 21460191 [PubMed - as supplied by publisher]
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Use of 1H NMR to measure intracellular metabolite levels during growth and asexual sporulation in Neurospora crassa.
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