Cloning of FPS gene and its role in regulation of alkaloid synthesis in Fritillaria thunbergii / 中草药

ABSTRACT

Objective: Farnesyl pyrophosphate synthase (FPS) gene of Fritillaria thunbergii was cloned and the correlation between the expression content of gene and alkaloid accumulation was analyzed to explore the role of FPS gene in the regulation of alkaloid synthesis and metabolism. Methods: In this study, the new bulbs and different tissues of Fritillaria thunbergii varieties "Narrow leaf" "Broad leaf" and "New Meiyuan" were used to clone the full-length sequence of FPS gene via RACE technology. RT-qPCR and HPLC-ELSD technology was used to determine the expression of FPS in 10 different developmental stages and the content of alkaloid (Peimine and Verticinone), respectively. Finally, the correlation was analyzed. Results: The full-length cDNA sequence of FPS gene was 1 629 bp. Open Reading Frame (ORF) was 1 056 bp and encoded 351 amino acids. Among them, the tissue specific expression trends of FPS gene were same, flowers had the highest expression level, followed by the new bulb. The results of correlation analysis showed thatthe content Peimine and Verticinone had significant positive correlation with the expression level of FPS gene in "Broad leaf" and "New Meiyuan" (The correlation coefficients were 0.289 and 0.613, 0.427 and 0.622, respectively);FPS gene expression and alkaloid content in different tissues had a positive correlation (The correlation coefficients were 0.057 and 0.476, 0.085 and 0.495, 0.375 and 0.432, respectively). Conclusion: The full-length sequence of FPS gene was successfully cloned in this study. The FPS gene involved in the regulation of alkaloid synthesis and metabolism.