Genome-Wide Identification and Expression Analysis of Flavonoid 3'-Hydroxylase Gene Family in Carthamus Tinctorius L.

Abstract ObjectiveThrough experiments and bioinformatic analysis clearly demonstrate considerable information about the genetic evolution of the flavonoid 3'-hydroxylase (F3'H) gene in Safflower and in plants.ResultsHere, we performed genome wide survey of safflower genome and identified a total of 22 CtF3'H enzyme encoding genes. Phylogenetic analyses revealed the classifications of these CtF3'Hs into nine subgroups demonstrating their evolutionary relationship. The distribution of the conserved protein motifs, and cis-regulatory units of CtF3'Hs indicated essential structure-to-function components leading to the final function of protein or its interactions. Furthermore, the results of digital expression analysis and the qRT-PCR pattern of 22 putative CtF3’H genes during different flowering stages suggested their requisite roles in safflower petal pigmentation. In addition, the fusions construct of plant expression vector pCAMBIA1302-GFP-CtF3’H5 in onion epidermal cells verified the subcellular localization of CtF3’H5 to the plasma membrane. Subsequently, the prokaryotic expression and western blot hybridization of CtF3’H5 resulted in a stable 50.3kD target protein. These results partly demonstrate the influence of F3'Hs on plants.ConclusionsIn this study, the results of digital expression and qRT-PCR analysis of 22 putative CtF3'H genes in different flowering stages indicate their essential role in safflower petal pigmentation. clearly demonstrates significant information on the genetic evolution of important enzyme-coding genes and will provide a pathway for future functional studies of F3'Hs in safflower as well as in plants.