摘要
Bulked-segregant analysis coupled with next-generation sequencing(BSA-seq) has emerged as an efficient tool for genetic mapping of single genes or major quantitative trait loci controlling(agronomic) traits of interest. However, such a mapping-by-sequencing approach usually relies on deep sequencing and advanced statistical methods. Application of BSA-Seq based on construction of reduced-representation libraries and allele frequency analysis permitted anchoring the barley pale-green(pg) gene on chromosome 3 HL. With further marker-assisted validation, pg was mapped to a 3.9 Mb physical-map interval. In the pg mutant a complete deletion of chlorophyllide a oxygenase(HvCAO) gene was identified.Because the product of this gene converts Chl a to Chl b, the pg mutant is deficient in Chl b.An independent Chl b-less mutant line M4437_2 carried a nonsynonymous substitution(F263 L) in the C domain of HvCAO. The study demonstrates an optimized pooling strategy for fast mapping of agronomically important genes using a segregating population.
Bulked-segregant analysis coupled with next-generation sequencing(BSA-seq) has emerged as an efficient tool for genetic mapping of single genes or major quantitative trait loci controlling(agronomic) traits of interest. However, such a mapping-by-sequencing approach usually relies on deep sequencing and advanced statistical methods. Application of BSA-Seq based on construction of reduced-representation libraries and allele frequency analysis permitted anchoring the barley pale-green(pg) gene on chromosome 3 HL. With further marker-assisted validation, pg was mapped to a 3.9 Mb physical-map interval. In the pg mutant a complete deletion of chlorophyllide a oxygenase(HvCAO) gene was identified.Because the product of this gene converts Chl a to Chl b, the pg mutant is deficient in Chl b.An independent Chl b-less mutant line M4437_2 carried a nonsynonymous substitution(F263 L) in the C domain of HvCAO. The study demonstrates an optimized pooling strategy for fast mapping of agronomically important genes using a segregating population.
基金
supported by the Young Elite Scientists Sponsorship Program by China Association for Science and Technology (2015QNRC001)
the National Natural Science Foundation of China (31370032)
the China Agriculture Research System (CARS-05)
the Agricultural Science and Technology Innovation Program
