Genome-wide identification of 1R-MYB transcription factors family and functional characterization of TrMYB130 under drought stresses in Trifolium repens (L.)

Abstract

White clover (Trifolium repens L.) is a high-quality leguminous forage, but its short rooting habit, poor transpiration tolerance, and drought tolerance, have become a key factor restricting its growth and cultivation. 1R-MYB transcription factors (TFs) are a significant subfamily of TFs in plants, playing a vital role in regulating plant responses to drought stress, however, knowledge about the role of 1R-MYB transcription factors in white clover is still limited. We identified 134 1R-MYB members, which were unevenly designated onto 16 chromosomes and divided phylogenetically into five subgroups. The members of the same subgroup had conserved motifs. Collinearity analysis revealed that segmental and tandem duplications significantly contributed to the expansion of the Tr1R-MYBs. Tr1R-MYBs promoter region enriched with potential drought cis-acting regulatory elements. The RT-qPCR results show that the five Tr1R-MYB genes (TrMYB41, TrMYB49, TrMYB94, TrMYB125, TrMYB130) have a certain degree of response under drought stress conditions but exhibited different expression profiles. Furthermore, subcellular localization analysis showed that the TrMYB130 protein is primarily located in the nucleus. Overexpression of this protein in transgenic Arabidopsis (Arabidopsis thaliana L.) was found to impair drought tolerance. Our findings will establish a basis for deeper investigation into the characteristics and functions of 1R-MYB TFs, as well as for employing genetic engineering techniques to improve white clover.