Cloning and expression analysis of MYB transcription factor gene involved in regulation of flavonoids biosynthesis in Carthamus tinctorius / 中草药

ABSTRACT

Objective To clone the MYB transcription factor gene in safflower, the sequence information and gene expression were analyzed to preliminarily identify the MYB transcription factor gene involved in regulation of flavonoid biosynthesis. Methods All the sequences of MYB transcription factor gene that was reported to involved in regulation of flavonoid biosynthesis were analyzed. The degenerate primers were designed to clone the core sequence. The full length of MYB transcription factor genes were cloned by RACE. The bioinformatics were used to analyze the sequences. The expression of MYB transcription factor genes in different tissues and different developmental stages of flower were analyzed by semi-quantitative PCR. Results Three candidate MYB transcription factors were cloned, named as CtFRMYB1, CtFRMYB2 and CtFRMYB3. The full lengths of the sequence were 1 223 bp, 1 080 bp and 1 348 bp, respectively. And the molecular weight were 17 878.15, 28 766.45 and 27 987.89, respectively. All three transcription factors have DNA binding domain and belong to the MYB family. Homologous analysis showed that CtFRMYB1 and CtFRMYB2 were closely related to AtMYB12. Expression analysis showed that CtFRMYB1 and CtFRMYB2 were only expressed in flowers, and the expression level was higher in flowering stage 3. Conclusion Three candidate MYB transcription factor genes involved in regulation of flavonoid biosynthesis (CtFRMYB1, CtFRMYB2 and CtFRMYB30) were successfully cloned. Two MYB transcription factor genes (CtFRMYB1 and CtFRMYB2) were identified by bioinformatics and expression analysis. These result laid a foundation for the molecular mechanism analysis of flavonoid biosynthesis.