Biologia Plantarum

, Volume 62, Issue 2, pp 250–260 | Cite as

Activation of polyketide synthase gene promoter in Cannabis sativa by heterologous transcription factors derived from Humulus lupulus

  • G. S. Duraisamy
  • A. K. Mishra
  • T. Kocábek
  • J. MatoušekEmail author
Original Papers


Cannabis sativa, an annual herbaceous plant, produce wide variety of secondary metabolites among which delta-9-tetrahydrocannabinol (THC) is the most important one. The dissection of biosynthetic pathway(s) of this compound and its regulation by transcription factors (TFs) is an important prerequisite for efficient biotechnological manipulation of its secondary metabolome. A polyketide synthase (PKS) of C. sativa catalyzes the first step of cannabinoid biosynthesis, leading to the biosynthesis of olivetolic acid. Cloning and analysis of PKS promoter based on online PLACE, PlantCARE, and Genomatix Matinspector professional databases, indicated that PKS promoter consisted of cis-elements such as TATA-box, CAAT-box, W-box, Myb-box, E-box, and P-box. Plant expression vector PKS::GUS was constructed in such a way that the ATG of the PKS gene was in the frame with the β-glucuronidase (GUS) coding region. Using a combinatorial transient GUS expression system in Nicotiana benthamania leaves, it was shown that heterologous TFs such as HlWRKY1, HlMYB3, HlWDR1 and HlbZIP1 from Humulus lupulus significantly activated PKS promoter. Moreover, Tombusvirus p19 core protein, which is known for silencing suppressor functions, acted in our combinatorial transient expression system as an enhancer of PKS promoter activity along with hop TFs. Our analyses suggested the involvement of the hop derived TFs (HlWRKY1, HlMYB3, HlWDR1 and HlbZIP1A) and p19 in the activation of PKS gene promoter, which could be used for the genetic manipulation of C. sativa to enhance the cannabinoid production.

Additional key words

β-glucuronidase bZIP1 MYB3 RT-qPCR Tombusvirus p19 core protein WDR1 WRKY1 



basic-leucine Zipper Domain 1


chalcone synthase






myeloblastosis proto-oncogene family of R2R3


polyketide synthase


35S cauliflower mosaic virus promoter


transcriptional factor


Tomato bushy stunt virus




Δ9-tetrahydrocannabinolic acid


WD repeat containing protein


transcription factor containing WRKYGQK motif




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Supplementary material

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Copyright information

© The Institute of Experimental Botany 2017

Authors and Affiliations

  • G. S. Duraisamy
    • 1
  • A. K. Mishra
    • 1
  • T. Kocábek
    • 1
  • J. Matoušek
    • 1
    Email author
  1. 1.Institute of Plant Molecular BiologyBiology Centre of the Academy of Sciences of the Czech RepublicČeské BudějoviceCzech Republic

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