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Fast Isogenic Mapping-by-Sequencing of Ethy1 Methanesulfonate-Induced Mutant Bulks
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Fast Isogenic Mapping-by-Sequencing of Ethy1 Methanesulfonate-Induced Mutant Bulks
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Journal Article
Fast Isogenic Mapping-by-Sequencing of Ethy1 Methanesulfonate-Induced Mutant Bulks
2012
Overview
Mapping-by-sequencing (or SHOREmapping) has revitalized the powerful concept of forward genetic screens in plants. However, as in conventional genetic mapping approaches, mapping-by-sequencing requires phenotyping of mapping populations established from crosses between two diverged accessions. In addition to the segregation of the focal phenotype, this introduces natural phenotypic variation, which can interfere with the recognition of quantitative phenotypes. Here, we demonstrate how mapping-by-sequencing and candidate gene identification can be performed within the same genetic background using only mutagen-induced changes as segregating markers. Using a previously unknown suppressor of mutants of like heterochromatin protein1 (Ihp1), which in its functional form is involved in chroma tin-mediated gene repression, we identified three closely linked ethy1 methanesulfonate-induced changes as putative candidates. In order to assess alíele frequency differences between such closely linked mutations, we introduced deep candidate resequencing using the new Ion Torrent Personal Genome Machine sequencing platform to our mutant identification pipeline and thereby reduced the number of causal candidate mutations to only one. Genetic analysis of two independent additional alíeles confirmed that this mutation was causal for the suppression of Ihp1.
Publisher
American Society of Plant Biologists
Subject
