Abstract:The amylose and amylopectin contents of 100 broomcorn millet germplasm resources were measured, and they were genotyped by molecular markers targeting the Waxy gene, the key factor modulating amylose synthesis. Some representative resources showing different amylose content were sequenced by Sanger approach. The amylose content in the collection ranged from 0 to 22.78%, with an average of 6.07%. 36% of germplasm resources were detected with amylase content less than 3.7%, including 24% detected without amylase. The amylopectin content ranged from 4.55% to 56.73%, with an average of 15.30%. The functional marker M5/R11 was able to effectively distinguish S0, S-15 and S0S-15 genotypes, and PCR products of markers int5Lf/R3 and M12/R12 were digested by endonucleases Acc I and EcoN I, enabling the identification of LC, LY and LF alleles. Nine genotypes at the Waxy locus were found in the collection. The genotype S0/Lc accounted for 33%, and S-15/LF accounted for 25%. The genotypes remained heterozygous (S0S-15/LF, S0S-15/LC and S0S-15/LYLF), and each of them accounted for 2%. A large number of new SNPs were found among landraces such as Gangu Heichanbei, Black millet, Qingyang Esiniu, as well as the varieties Jinshu 9 and Chimi 2. These SNPs serve a potential in future gene function analysis and innovative breeding in broomcorn millet. Collectively, this study provided theoretical basis for molecular breeding and quality improvement in broomcorn millet.