In order to widen gene germplasm for kernel hardness in triticale, 60 synthetic hexaploid triticales were tested by the single kernel characterization system(SKCS) and secaloindoline alleles were identified by sequencing. Phenot^ing showed that frequencies of soft, mixed and hard genotypes were 43.3%, 48.3%, and 8.4%, respectively. Genotyping identified three known secaloindoline-a alleles and four known secaloindoline-b alleles. Three new Sina-Rl alleles were designated Sina-Rld, Sina-Rle and Sina-Rlf. Compared to Sina-Dlc, Sina-Rld showed four point mutations causing changes in four amino acids, Sina-Rle had one point mutation causing an alanine to glycine substitution, and Sina-Rlf possessed five point mutations but produced the same amino add sequence as Sina-Rld. Two new Sinb-Rl alleles were discovered and designated Sinb-Rle and Sinb-Rl/. Compared to Sinb-Rla, Sinb-Rle possessed a triplet-code insertion and four point mutations causing a cysteine insertion and two amino acid substitutions, and Sinb-Rl/possessed three point mutations causing a cysteine insertion and a change of arginine to glycine.Association of hardness index with secaloindoline alleles indicated t±iat SKCS of the Sina-Rld genotype was significantly lower than that of Sina-Rle, and Sinb-Rle was significantly lower than that of the Sinb-Rld genotype. A total of eight allelic combinations of secaloindoline genes were identified; Sma-Rld/Sinb-Rle and Sina-Rle/Sinb-Rld were relatively prevalent in the triticales surveyed. The results provide valuable information for further use of these germplasms in triticale breeding program due to the diverse polymorphism in secaloindoline genes.
