Genetic Transformation of Pentalinon andrieuxii Tissue Cultures

Methods Mol Biol. 2018:1815:475-492. doi: 10.1007/978-1-4939-8594-4_33.

Abstract

Pentalinon andrieuxii is a species used in Mayan traditional medicine due to its biological properties. Recent studies indicate that it produces a pentacyclic triterpene-denominated betulinic acid, which presents various biological activities: antibacterial, antifungal, antiplasmodial, anti-inflammatory, antimalarial, anticancer, leishmanicidal, and antiviral, as well as steroids and sterols with leishmanicidal properties. A recent study also reported the presence of urechitol A and B in the roots; these are secondary metabolites whose biochemical function is as yet unknown. This plant therefore represents a natural source of metabolites with potential application in the pharmaceutical industry. In this chapter, a protocol is described for obtaining transgenic plants, at the reporter gene of the β-glucuronidase (GUS) via Agrobacterium tumefaciens from hypocotyl and root explants. The protocol established herein could be employed for the manipulation of the genes involved in the biosynthesis of isoprenoids or secondary metabolites of interest. To our knowledge, this is the first report of stable transformation of Pentalinon andrieuxii via Agrobacterium tumefaciens.

Keywords: Agrobacterium tumefaciens; Pentalinon andrieuxii; Stable genetic transformation; β-glucuronidase gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptation, Physiological
  • Agrobacterium tumefaciens / metabolism
  • Apocynaceae / genetics*
  • Culture Media / chemistry
  • Genes, Reporter
  • Germination / drug effects
  • Glucuronidase / metabolism
  • Hypocotyl / growth & development
  • Kanamycin / pharmacology
  • Plant Shoots / physiology
  • Plasmids / metabolism
  • Polymerase Chain Reaction
  • Seeds / physiology
  • Tissue Culture Techniques / methods*
  • Transformation, Genetic*

Substances

  • Culture Media
  • Kanamycin
  • Glucuronidase