Heat Shock Improves Random Spore Analysis in Diverse Strains of Saccharomyces cerevisiae
Random spore analysis (RSA) is a classic method in yeast genetics that allows high-throughput purification of recombinant haploid spores following specific crosses. RSA typically involves a number of steps to induce sporulation, purge vegetative cells that fail to sporulate, and disrupt the ascus wa...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-12-01
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| Series: | Frontiers in Genetics |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fgene.2020.597482/full |
| _version_ | 1819275466778869760 |
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| author | Molly K. Burke Kaitlin M. McHugh Ian C. Kutch |
| author_facet | Molly K. Burke Kaitlin M. McHugh Ian C. Kutch |
| author_sort | Molly K. Burke |
| collection | DOAJ |
| description | Random spore analysis (RSA) is a classic method in yeast genetics that allows high-throughput purification of recombinant haploid spores following specific crosses. RSA typically involves a number of steps to induce sporulation, purge vegetative cells that fail to sporulate, and disrupt the ascus walls of sporulated cells to release haploid spores. These steps generally require expensive chemicals and/or enzymes that kill diploid cells but have few effects on spores. In the fission yeast Schizosaccharomcyes pombe, heat shock has been reported as an effective addition to RSA protocols, but to our knowledge heat shock has not been used for this purpose in the budding yeast Saccharomyces cerevisiae. Here, we evaluate the effects of heat shock on vegetative and sporulated cultures of four diverse yeast strains: a European wine strain (DBVPG6765), a Japanese sake strain (Y12), a West African palm wine strain (DBVPG6044) and a North American strain isolated from the soil beneath an oak tree (YPS128). We characterize this phenotype under multiple combinations of temperature and incubation time, and find specific conditions that lead to the exclusion of vegetative cells and an enrichment in spores, which differ by strain. We also collected genome sequence data from a recombinant population that experienced multiple rounds of RSA, including one round with a heat shock treatment. These data suggest that when incorporated into an RSA protocol, heat shock leads to increased genetic diversity among the cells that survive and mate. Ultimately, our work provides evidence that short heat treatments can improve existing RSA protocols, though in a strain-specific manner. This result informs applications of high-throughput RSA protocols, such as QTL mapping and experimental evolution research. |
| first_indexed | 2024-12-23T23:24:47Z |
| format | Article |
| id | doaj.art-bd86a010ada24487af4b315537215fb2 |
| institution | Directory Open Access Journal |
| issn | 1664-8021 |
| language | English |
| last_indexed | 2024-12-23T23:24:47Z |
| publishDate | 2020-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Genetics |
| spelling | doaj.art-bd86a010ada24487af4b315537215fb22022-12-21T17:26:15ZengFrontiers Media S.A.Frontiers in Genetics1664-80212020-12-011110.3389/fgene.2020.597482597482Heat Shock Improves Random Spore Analysis in Diverse Strains of Saccharomyces cerevisiaeMolly K. BurkeKaitlin M. McHughIan C. KutchRandom spore analysis (RSA) is a classic method in yeast genetics that allows high-throughput purification of recombinant haploid spores following specific crosses. RSA typically involves a number of steps to induce sporulation, purge vegetative cells that fail to sporulate, and disrupt the ascus walls of sporulated cells to release haploid spores. These steps generally require expensive chemicals and/or enzymes that kill diploid cells but have few effects on spores. In the fission yeast Schizosaccharomcyes pombe, heat shock has been reported as an effective addition to RSA protocols, but to our knowledge heat shock has not been used for this purpose in the budding yeast Saccharomyces cerevisiae. Here, we evaluate the effects of heat shock on vegetative and sporulated cultures of four diverse yeast strains: a European wine strain (DBVPG6765), a Japanese sake strain (Y12), a West African palm wine strain (DBVPG6044) and a North American strain isolated from the soil beneath an oak tree (YPS128). We characterize this phenotype under multiple combinations of temperature and incubation time, and find specific conditions that lead to the exclusion of vegetative cells and an enrichment in spores, which differ by strain. We also collected genome sequence data from a recombinant population that experienced multiple rounds of RSA, including one round with a heat shock treatment. These data suggest that when incorporated into an RSA protocol, heat shock leads to increased genetic diversity among the cells that survive and mate. Ultimately, our work provides evidence that short heat treatments can improve existing RSA protocols, though in a strain-specific manner. This result informs applications of high-throughput RSA protocols, such as QTL mapping and experimental evolution research.https://www.frontiersin.org/articles/10.3389/fgene.2020.597482/fullrandom spore analysisheat shocksporulationgenomicsmultiparent mapping population |
| spellingShingle | Molly K. Burke Kaitlin M. McHugh Ian C. Kutch Heat Shock Improves Random Spore Analysis in Diverse Strains of Saccharomyces cerevisiae Frontiers in Genetics random spore analysis heat shock sporulation genomics multiparent mapping population |
| title | Heat Shock Improves Random Spore Analysis in Diverse Strains of Saccharomyces cerevisiae |
| title_full | Heat Shock Improves Random Spore Analysis in Diverse Strains of Saccharomyces cerevisiae |
| title_fullStr | Heat Shock Improves Random Spore Analysis in Diverse Strains of Saccharomyces cerevisiae |
| title_full_unstemmed | Heat Shock Improves Random Spore Analysis in Diverse Strains of Saccharomyces cerevisiae |
| title_short | Heat Shock Improves Random Spore Analysis in Diverse Strains of Saccharomyces cerevisiae |
| title_sort | heat shock improves random spore analysis in diverse strains of saccharomyces cerevisiae |
| topic | random spore analysis heat shock sporulation genomics multiparent mapping population |
| url | https://www.frontiersin.org/articles/10.3389/fgene.2020.597482/full |
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