High-throughput genotyping by whole-genome resequencing.
The next-generation sequencing technology coupled with the growing number of genome sequences opens the opportunity to redesign genotyping strategies for more effective genetic mapping and genome analysis. We have developed a high-throughput method for genotyping recombinant populations utilizing whole-genome resequencing data generated by the Illumina Genome Analyzer. A sliding window approach is designed to collectively examine genome-wide single nucleotide polymorphisms for genotype calling and recombination breakpoint determination. Using this method, we constructed a genetic map for 150 rice recombinant inbred lines with an expected genotype calling accuracy of 99.94% and a resolution of recombination breakpoints within an average of 40 kb. In comparison to the genetic map constructed with 287 PCR-based markers for the rice population, the sequencing-based method was approximately 20x faster in data collection and 35x more precise in recombination breakpoint determination. Using the sequencing-based genetic map, we located a quantitative trait locus of large effect on plant height in a 100-kb region containing the rice "green revolution" gene. Through computer simulation, we demonstrate that the method is robust for different types of mapping populations derived from organisms with variable quality of genome sequences and is feasible for organisms with large genome sizes and low polymorphisms. With continuous advances in sequencing technologies, this genome-based method may replace the conventional marker-based genotyping approach to provide a powerful tool for large-scale gene discovery and for addressing a wide range of biological questions.
National Center for Gene Research and Institute of Plant Physiology and Ecology, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200233, China.
Xuehui
X
Huang
Qi
Q
Feng
Qian
Q
Qian
Qiang
Q
Zhao
Lu
L
Wang
Ahong
A
Wang
Jianping
J
Guan
Danlin
D
Fan
Qijun
Q
Weng
Tao
T
Huang
Guojun
G
Dong
Tao
T
Sang
Bin
B
Han
D000465:Algorithms; D001483:Base Sequence; D002874:Chromosome Mapping; D003198:Computer Simulation; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D018745:Genome, Plant / Q000235:genetics*; D005838:Genotype; D012275:Oryza / Q000235:genetics* / Q000254:growth & development; D020641:Polymorphism, Single Nucleotide; D040641:Quantitative Trait Loci / Q000235:genetics; D011995:Recombination, Genetic; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods*
D018744:DNA, Plant