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Background  Anthocyanins are plants secondary metabolites important for plant adaptation to severe environments and potentially beneficial to human health. Purple colour of barley grain is caused by the pigments synthesized in pericarp. One or two genes determine the trait. One of them is  Ant2  mapped on chromosome 2HL and is known to encode transcription factor (TF) with a bHLH domain. In plants, bHLH regulates anthocyanin biosynthesis together with TF harboring an R2R3-MYB domain. In wheat, the R2R3-MYBs responsible for purple colour of grain pericarp are encoded by the homoallelic series of the  Pp-1  genes that were mapped on the short arms of chromosomes 7. In barley, in orthologous positions to wheat’s  Pp-1 , the  Ant1  gene determining red colour of leaf sheath has been mapped. In the current study, we tested whether  Ant1  has pleiotropic effect not only on leaf sheath colour but also on pericarp pigmentation.    Results  А set of near isogenic lines (NILs) carrying different combinations of alleles at the  Ant1  and  Ant2  loci was created using markers-assisted backcrossing approach. The dominant alleles of both the  Ant1  and  Ant2  genes are required for anthocyanin accumulation in pericarp. A qRT-PCR analysis of the  Ant  genes in lemma and pericarp of the NILs revealed that some reciprocal interaction occurs between the genes. Expression of each of the two genes was up-regulated in purple-grained line with dominant alleles at the both loci. The lines carrying dominant allele either in the  Ant1  or in the  Ant2  locus were characterized by the decreased level of expression of the dominant gene and scant activity of the recessive one. The  Ant1  and  Ant2  expression was barely detected in uncolored line with recessive alleles at both loci. The anthocyanin biosynthesis structural genes were differently regulated:  Chs ,  Chi ,  F3h ,  Dfr  were transcribed in all lines independently on allelic state of the  Ant1  and  Ant2  genes, whereas  F3’h  and  Ans  were activated in presence on dominant alleles of the both regulatory genes.    Conclusions  The R2R3-MYB-encoding counterpart ( Ant1 ) of the regulatory  Ant2  gene was determined for the first time. The dominant alleles of both of them are required for activation of anthocyanin synthesis in barley lemma and pericarp. The R2R3-MYB + bHLH complex activates the synthesis via affecting expression of the  F3’h  and  Ans  structural genes. In addition, positive regulatory loop between  Ant1  and  Ant2  was detected. Earlier the interaction between the anthocyanin biosynthesis regulatory genes has been revealed in dicot plant species only. Our data demonstrated that the regulatory mechanism is considered to be more common for plant kingdom than it has been reported so far.    Electronic supplementary material  The online version of this article (10.1186/s12870-019-1638-9) contains supplementary material, which is available to authorized users.

bmc plant biology

Purple-grained barley (Hordeum vulgare L.): marker-assisted development of NILs for investigating peculiarities of the anthocyanin biosynthesis regulatory network