Iwate Biotechnology Research Center, Kitakami, Iwate, Japan.
PLoS One. 2013 Jul 10;8(7):e68529. doi: 10.1371/journal.pone.0068529. Print 2013.
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.
基因组测序技术的进步使研究人员和育种者能够快速将表型变异与基因组序列差异联系起来。我们最近利用下一代测序技术开发了 MutMap 方法，该方法通过对候选突变体与亲本系杂交产生的突变 F2 后代的混合 DNA 进行全基因组重测序，可快速鉴定水稻突变体的因果核苷酸变化。在这里，我们描述了 MutMap+，这是 MutMap 的一种通用扩展，通过比较自交产生的 M2 杂合个体的 M3 代突变体和野生型后代的 bulked DNA 的 SNP 频率来鉴定因果突变。值得注意的是，MutMap+不需要突变体与野生型亲本系之间的人工杂交。因此，该方法适用于鉴定导致早期发育致死、不育或普遍阻碍杂交的突变。此外，MutMap+对于那些难以进行人工杂交的作物的基因分离也具有潜在的应用价值。
