期刊
FRONTIERS IN PLANT SCIENCE
卷 11, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fpls.2020.01249
关键词
wheat exome capture; varBScore; gene cloning; bulked segregant analysis; CRISPR; Cas9; yellow-green leaf mutant
资金
- National Transgenic Research Project [2016ZX08009001-001-004]
- National Key Research and the Development Program of China [2016YFD0101001]
- Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences (CAAS)
- Tianshan Innovation Team Plan [2020D14002]
The discovery of functional genes underlying agronomic traits is of great importance for wheat improvement. Here we designed a new wheat exome capture probe panel based on IWGSC RefSeq v1.0 genome sequence information and developed an effective algorithm, varBScore, that can sufficiently reduce the background noise in gene mapping and identification. An effective method, termedbulkedsegregantexome capturesequencing (BSE-Seq) for identifying causal mutations or candidate genes was established by combining the use of a newly designed wheat exome capture panel, sequencing of bulked segregant pools from segregating populations, and the robust algorithm varBScore. We evaluated the effectiveness of varBScore on SNP calling using the published dataset for mapping and cloning the yellow rust resistance geneYr7in wheat. Furthermore, using BSE-Seq, we rapidly identified a wheat yellow leaf mutant gene,ygl1, in an ethyl methanesulfonate (EMS) mutant population and found that a single mutation of G to A at 921 position in the wild typeYGL1gene encoding magnesium-chelatase subunit chlI caused the leaf yellowing phenotype. We further showed that mutation ofYGL1through CRISPR/Cas9 gene editing led to a yellow phenotype on the leaves of transgenic wheat, indicating that ygl1 is the correct causal gene responsible for the mutant phenotype. In summary, our approach is highly efficient for discovering causal mutations and gene cloning in wheat.
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