Molecular cloning and characterization of the promoter of aldehyde dehydrogenase gene from Artemisia annua

Biotechnol Appl Biochem. 2017 Nov;64(6):902-910. doi: 10.1002/bab.1520. Epub 2017 Sep 7.

Abstract

In recent years, although several related genes had been cloned and characterized, the role of aldehyde dehydrogenase 1 (ALDH1), the newly cloned gene involved in artemisinin biosynthesis pathway, is still not clear. In this study, a 2,100-bp ALDH1 promoter region fused with GUS reporter gene was stably transferred into Arabidopsis thaliana. Histochemical staining showed the methyl jasmonate (MeJA) and wounding treatment induced the GUS gene expression specifically in the trichomes of transgenic A. thaliana, consistent with the results that the expression level of ALDH1 gene was increased in the A. annua under MeJA and wounding treatments. Two RAA motifs (AP2/ERF binding site) but no W box (WRKY binding site) motif were identified in the ALDH1 promoter by the analysis through PLACE and plantCARE. Through the dual luciferase reporter assay, we revealed that both AaORA and AaERF2, rather than AaWRKY1, could activate the expression of ALDH1 promoter. Our study shed light on the in-depth understanding of the role of ALDH1 in artemisinin biosynthesis.

Keywords: ALDH1; Artemisia annua L.; cis-acting regulatory elements; promoter activity.

MeSH terms

  • Aldehyde Dehydrogenase 1 Family
  • Artemisia annua / enzymology*
  • Cloning, Molecular
  • DNA, Plant / genetics
  • Isoenzymes / genetics*
  • Promoter Regions, Genetic / genetics*
  • Retinal Dehydrogenase / genetics*

Substances

  • DNA, Plant
  • Isoenzymes
  • Aldehyde Dehydrogenase 1 Family
  • Retinal Dehydrogenase