Marked Neurospora crassa strains for competition experiments and Bayesian methods for fitness estimates
| Autor: | Eric U. Selker, Ilkka Kronholm, Tarmo Ketola, Kevin J McNaught, Tereza Ormsby |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
Genetics 0303 health sciences Mating type Experimental evolution biology evoluutiobiologia Crassa Locus (genetics) QH426-470 biology.organism_classification 010603 evolutionary biology 01 natural sciences DNA extraction high resolution melting Neurospora crassa competitive fitness 03 medical and health sciences fungi experimental evolution Allele sienet Gene 030304 developmental biology |
| Zdroj: | G3: Genes, Genomes, Genetics, Vol 10, Iss 4, Pp 1261-1270 (2020) |
| DOI: | 10.1101/736645 |
| Popis: | The filamentous fungusNeurospora crassa, a model microbial eukaryote, has a life cycle with many features that make it suitable for studying experimental evolution. However, it has lacked a general tool for estimating relative fitness of different strains in competition experiments. To remedy this need, we constructedN. crassastrains that contain a modifiedcsr-1locus and developed an assay for detecting the proportion of the marked strain using a post PCR high resolution melting assay. DNA extraction from spore samples can be performed on 96-well plates, followed by a PCR step, which allows many samples to be processed with ease. Furthermore, we suggest a Bayesian approach for estimating relative fitness from competition experiments that takes into account the uncertainty in measured strain proportions. We show that there is a fitness effect of the mating type locus, as mating typemat ahas a higher competitive fitness thanmat A. Thecsr-1* marker also has a small fitness effect, but is still a suitable marker for competition experiments. As a proof of concept, we estimate the fitness effect of theqde-2mutation, a gene in the RNA interference pathway, and show that its competitive fitness is lower than what would be expected from its mycelial growth rate alone. |
| Databáze: | OpenAIRE |
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