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Triticum boeoticum  Boiss (A b A b , 2n = 2x = 14) is one of the sources of the blue grain trait controlled by  blue aleurone layer 2  ( Ba2 ). However, the underlying genes have not been cloned. In this study, a transcriptomic comparison between a blue-grained wheat- T. boeoticum  substitution line and its wheat parent identified 41 unigenes related to anthocyanin biosynthesis and 29 unigenes related to transport. The bHLH transcription factor gene  TbMYC4A  showed a higher expression level in the blue-grained substitution line.  TbMYC4A  contained the three characteristic bHLH transcription factor domains (bHLH-MYC_N, HLH and ACT-like) and clustered with genes identified from other wheat lines with the blue grain trait derived from other Triticeae species.  TbMYC4A  overexpression confirmed that it was a functional bHLH transcription factor. The analysis of a  TbMYC4A- specific marker showed that the gene was also present in  T. boeoticum  and  T. monococcum  with blue aleurone but absent in other Triticeae materials with white aleurone. These results indicate that  TbMYC4A  is a candidate gene of  Ba2  controlling the blue aleurone trait. The isolation of  TbMYC4A  is helpful for further clarifying the genetic mechanism of the blue aleurone trait and is of great significance for breeding blue-grained wheat varieties.

TbMYC4A Is a Candidate Gene Controlling the Blue Aleurone Trait in a Wheat-Triticum boeoticum Substitution Line