Journal
PLOS ONE
Volume 8, Issue 7, Pages -Publisher
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0068529
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Funding
- Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN)
- Ministry of Agriculture, Forestry, and Fisheries of Japan [PMI-0010]
- Ministry of Education, Cultures, Sports and Technology, Japan [23113009]
- JSPS KAKENHI [24248004]
- Gatsby Charitable Foundation
- Grants-in-Aid for Scientific Research [22510217, 12F02212, 23580065] Funding Source: KAKEN
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Advances in genome sequencing technologies have enabled researchers and breeders to rapidly associate phenotypic variation to genome sequence differences. We recently took advantage of next-generation sequencing technology to develop MutMap, a method that allows rapid identification of causal nucleotide changes of rice mutants by whole genome resequencing of pooled DNA of mutant F2 progeny derived from crosses made between candidate mutants and the parental line. Here we describe MutMap+, a versatile extension of MutMap, that identifies causal mutations by comparing SNP frequencies of bulked DNA of mutant and wild-type progeny of M3 generation derived from selfing of an M2 heterozygous individual. Notably, MutMap+ does not necessitate artificial crossing between mutants and the wild-type parental line. This method is therefore suitable for identifying mutations that cause early development lethality, sterility, or generally hamper crossing. Furthermore, MutMap+ is potentially useful for gene isolation in crops that are recalcitrant to artificial crosses.
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