[Cloning and expression analysis of F3'H and quantification of downstream products in Chrysanthemum morifolium under flooding stress]

Zhongguo Zhong Yao Za Zhi. 2018 Jan;43(1):52-57. doi: 10.19540/j.cnki.cjcmm.20171030.008.
[Article in Chinese]

Abstract

To investigate the effects of the expression of flavonoid 3' hydroxylase gene (F3'H) and active ingredients in Chrysanthemum morifolium under flooding stress, we cloned F3'H from Hangju (temporarily named CmF3'H) and conducted bioinformatics analysis. During the flower bud differentiation stage, we flooded the Ch. morifolium and then used the Real-time PCR to detect the relative expression of CmF3'H; Finally, active ingredients of the inflorescence were measured by HPLC.The sequencing results showed that 1 562 bp sequence was acquired with the largest open reading frame of 1 527 bp, which encoded 508 amino acids. The phylogenetic tree found that CmF3'H was highly homologous to other species of Compositae. Real-time PCR results showed that CmF3'H had a significant response to flooding stress and had the highest expression level after flooding for 24 h, which was about 9 times as that of the control group. The results of HPLC showed that luteolin and luteoloside, the downstream products catalyzed by the F3'H, were significantly higher than those in the control group. It was also found that the contents of chlorogenic acid and 3,5-O-di-caffeoylquinic acid were also significantly higher than those of the control group. Therefore, Ch. morifolium regulates the synthesis of downstream products by regulating the expression of CmF3'H in the flavonoid synthesis pathway under flooding stress, thereby responding to flooding stress. The flooding stress during flower bud differentiation can significantly enhance the accumulation of active ingredients.

Keywords: Chrysanthemum morifolium; F3'H; active ingredients; clone; flavonoids; flooding stress.

MeSH terms

  • Chrysanthemum / enzymology*
  • Chrysanthemum / genetics
  • Cloning, Molecular
  • Cytochrome P-450 Enzyme System / genetics*
  • Floods*
  • Gene Expression Regulation, Plant
  • Glucosides / biosynthesis
  • Luteolin / biosynthesis
  • Phylogeny
  • Plant Proteins / genetics*
  • Stress, Physiological*

Substances

  • Glucosides
  • Plant Proteins
  • Cytochrome P-450 Enzyme System
  • luteolin-7-glucoside
  • flavonoid 3'-hydroxylase
  • Luteolin