Molecular cloning and sequence variance analysis of the TEOSINTE BRANCHED1 (TB1) gene in bermudagrass [Cynodon dactylon (L.) Pers]

TEOSINTE BRANCHED1 (TB1) encodes a TEOSINTE BRANCHED1, CYCLOIDEA, and PROLIFERATING CELL FACTOR (TCP) transcription factor that represses axillary bud outgrowth and lateral branch formation in plants. Previous studies have elucidated the essential tillering regulatory roles of TB1 in many grasses, including maize and rice; however, the functions of TB1 in turf grasses remain unclear. In this study, we cloned the CdTB1 gene from bermudagrass, an important turfgrass species, and characterized the transactivation function of the CdTB1 protein. Sequencing the CdTB1 gene locus in a mini-core germplasm collection of Chinese bermudagrasses led to the successful identification of 66 SNP and 2 indel mutations in the protein-coding region as well as 28 SNP and 11 indel mutations in the promoter region. Interestingly, mutations in the C-terminal transactivation domain of the CdTB1 protein had no significant influence on the transactivation activity, whereas a novel 335-bp insertion mutation located in the promoter region could significantly increase the expression of the CdTB1 gene. Furthermore, wild accessions of bermudagrass harboring the novel insertion mutation were found to have significantly reduced tillers compared with other accessions, suggesting a negative correlation between the mutation and tillering. The results of this study not only expanded our knowledge of TB1 gene expression regulation but also provided possible molecular markers to breed cultivars of turf and forage grasses with specific architectural features.