Construction of a high-density mutant population of Chinese cabbage facilitates the genetic dissection of agronomic traits

Chinese cabbage(Brassica rapa ssp.pekinensis)is an economically important vegetable crop throughout the world,especially in Asia.High-quality genome sequences are available for Chinese cabbage,but gene functional studies remain challenging.To promote functional genomic studies of Chinese cabbage,we generated an ethyl methane sulfonate(EMS)mutant population of～8000 M2 plants using the double haploid inbred line A03 as the parent.The genome of A03 was sequenced and used as a reference for high-throughput functional characterization of gene mutations at the whole-genome level.A total of 300 M2 to M5 EMS mutants were phenotypically screened and then sequenced,revealing 750 629 SNPs and 46 272 InDel mutations that cover 98.27％of all predicted genes in the A03 genome.A forward-genetics approach was successfully used to identify two genes with chloroplast-related functions that are responsible for the yellow leaf mutant trait.A reverse-genetics approach was also used to identify associations between mutations in five genes of the glucosinolate biosynthetic pathway and variations in glucosinolate content of the mutant plants.In addition,we built the Chinese cabbage EMS mutation database(CCEMD,www.bioinformaticslab.cn/EMSmutation/home)to increase the usability of this mutant population resource.In summary,we performed large-scale screening of a heading Chinese cabbage EMS mutant collection at the phenotypic and genotypic levels,which will facilitate gene mining of Chinese cabbage and might also be useful for the study of other Brassica crops.