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The gibberellin-responsive element binding factor (GRAS) family of proteins plays an important role in the transcriptional regulation of plant development and hormone signaling. To date, there are no reports on GRAS family proteins expressed in  Salvia miltiorrhiza . In this study, 28 ESTs that contained the  GRAS  domain were identified from a  S .  miltiorrhiza  cDNA library. Of these, full-length sequences of five genes were cloned and sequence analysis indicated that all five proteins contain only one GRAS domain and therefore, belong to the GRAS family. The five genes were designated  S .  miltiorrhiza GRAS1-5  ( SmGRAS1-5 ), which belong to groups I ( SmGRAS2  and  SmGRAS4 ), II ( SmGRAS3 ), III ( SmGRAS1 ), and VIII ( SmGRAS5 ) respectively. Additionally,  SmGRAS1-5  have different expression patterns in the reed head, stems, leaves, flowers, and roots of  S .  miltiorrhiza . In this study, the expression of  SmGRAS1-5  was sensitive to Gibberellin (GA) stress and that of  SmGRAS1 ,  SmGRAS4  and  SmGRAS5  was sensitive to Ethephon (Eth) stress respectively. Moreover,  S .  miltiorrhiza  copalyl diphosphate synthases 1 ( SmCPS1 ) and  S .  miltiorrhiza  kaurene synthase like 1 ( SmKSL1 ), which are two key enzymes gene in the diterpenoid biosynthesis pathway, were also response to GA and Eth stress. In addition, Dihydrotanshinone (DT-I) and Tanshinone I (T-I) content were enhanced by GA and Eth stress, Tanshinone IIA (T-IIA) content was increased by GA stress, and the accumulation of Cryptotanshinone (CT) was insensitive to both GA and Eth stress. Together, these results provide insights into functional conservation and diversification of SmGRASs and are useful information for further elucidating SmGRAS functions.

Molecular cloning and characterization of five SmGRAS genes associated with tanshinone biosynthesis in Salvia miltiorrhiza hairy roots