Screening and kinetic studies of catharanthine and ajmalicine accumulation and their correlation with growth biomass in Catharanthus roseus hairy roots

Pharm Biol. 2016 Oct;54(10):2033-43. doi: 10.3109/13880209.2016.1140213. Epub 2016 Mar 17.

Abstract

Context Catharanthus roseus (L.) G. Don (Apocynaceae) is still one of the most important sources of terpene indole alkaloids including anticancer and hypertensive drugs as vincristine and vinblastine. These final compounds have complex pathway and many enzymes are involved in their biosynthesis. Indeed, ajmalicine and catharanthine are important precursors their increase can lead to enhance levels of molecules of interest. Objective This study aims at selecting the highest yield of hairy root line(s) and at identifying best times for further treatments. We study kinetics growth and alkaloids (ajmalicine and catharanthine) accumulation of three selected hairy root lines during the culture cycle in order to determine the relationship between biomass production and alkaloids accumulation. Materials and methods Comparative analysis has been carried out on three selected lines of Catharanthus roseus hairy roots (LP10, LP21 and L54) for their kinetics of growth and the accumulation of ajamalicine and catharanthine, throughout a 35-day culture cycle. The methanolic extract for each line in different times during culture cycle is analyzed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Results Maximum accumulation of the alkaloids is recorded for LP10 line in which the peak of ajmalicine and catharanthine accumulation reached to 3.8 and 4.3 mg/g dry weight (DW), respectively. This increase coincides with an exponential growth phase. Discussion and conclusion Our results suggest that the evolution of accumulation of ajmalicine and catharanthine are positively correlated with the development of the biomass growth. Significantly, for LP10 line the most promising line to continue optimizing the production of TIAs. Additionally, the end of exponential phase remains the best period for elicitor stimuli.

Keywords: Agrobacterium rhizogenes; growth kinetics; medicinal plant; secondary metabolites; terpene indole alkaloids.

MeSH terms

  • Agrobacterium / genetics
  • Agrobacterium / physiology*
  • Biomass*
  • Catharanthus / genetics
  • Catharanthus / growth & development
  • Catharanthus / metabolism*
  • Catharanthus / microbiology
  • Chromatography, Liquid
  • Gene Expression Regulation, Plant
  • Host-Pathogen Interactions
  • Kinetics
  • Methanol / chemistry
  • Phytotherapy
  • Plant Roots / growth & development
  • Plant Roots / metabolism*
  • Plant Roots / microbiology
  • Plants, Medicinal
  • Secologanin Tryptamine Alkaloids / metabolism*
  • Solvents / chemistry
  • Tandem Mass Spectrometry
  • Transformation, Genetic
  • Vinca Alkaloids / metabolism*

Substances

  • Secologanin Tryptamine Alkaloids
  • Solvents
  • Vinca Alkaloids
  • raubasine
  • catharanthine
  • Methanol