MutMap + in Nipponbare late. Oryza sativa Japonica Group

We applied MutMap+ method to identify the causal nucleotide changes of rice mutants by whole genome re-sequencing .The mutants was first crossed with Wild type. F1 plant self-pollinated, and the F2 progeny were grown for investigating heading date. We observed segregation between wild type and mutant phenotype in F2 progeny. We next made two bulks of DNA; wild type bulk of 20 individuals from the wild type phenotype and mutant bulk of 20 individuals from the mutant phenotype. The two bulks of DNA were separately sequenced to generate illumina HiSeq 2000 150bp paired end reads.

本研究采用MutMap+定位法，通过全基因组重测序（whole genome re-sequencing）鉴定水稻突变体的致因核苷酸变异。首先将突变体与野生型（Wild type）进行杂交，所得F1代植株自花授粉，随后种植F2代群体以调查抽穗期（heading date）。在F2代群体中观察到野生型与突变体表型的性状分离。随后构建两份DNA混池：一份由20株表现为野生型表型的单株提取DNA混合得到，另一份由20株表现为突变体表型的单株提取DNA混合得到。对这两份DNA混池分别进行测序，获取illumina HiSeq 2000平台的150bp双端测序读段（paired end reads）。