Agrobacterium tumefaciens-Mediated Stable Transformation of Daucus carota

Methods Mol Biol. 2020:2083:313-320. doi: 10.1007/978-1-4939-9952-1_24.

Abstract

Daucus carota L. (carrot) is one of the ten most important vegetables cultivated and consumed worldwide and is a main source of provitamin A. Carrot storage root is rich in dietary fiber, antioxidants, and other nutrients but especially in carotenoids. It has been also used as plant model for studding embryogenesis, as well as the genetic and genomic evolution of carrots and for carotenoid synthesis regulation, among others. Research in carrot often needs genetic transformation. Here we describe a step-by-step protocol on the nuclear and stable transformation of carrot through Agrobacterium tumefaciens and somatic embryogenesis in vitro culture. Somatic embryos, induced by supplementation of Murashige-Skoog medium with the 2,4D hormone, develop into seedlings after 6 months approximately when plants are ready to be transferred to a greenhouse. The protocol has over 85% of transformation efficiency.

Keywords: 2,4-dichlorophenoxiacetic acid; Agrobacterium tumefaciens; Carrot; In vitro culture; Somatic embryogenesis; Transformation protocol.

MeSH terms

  • Agrobacterium tumefaciens / genetics*
  • Daucus carota / genetics*
  • Gene Expression Regulation, Plant*
  • Phenotype
  • Plant Development / genetics
  • Plant Somatic Embryogenesis Techniques
  • Plants, Genetically Modified
  • Transformation, Genetic*