Chapter 24
Agrobacterium tumefaciens-Mediated Stable
Transformation of Daucus carota
Christian Gonzalez-Calquin and Claudia Stange
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.
Key words Carrot, Transformation protocol, 2,4-dichlorophenoxiacetic acid, Somatic embryogenesis, Agrobacterium tumefaciens, In vitro culture
1 Introduction
Carrot (Daucus carota L., 2n ¼ 18) belongs to the Umbelliferae
group together with parsley (Petroselinum hortense), anise (Pimpinella anisum), celery (Apium graveolens), caraway (Carum carvi),
dill (Anethum graveolens), and other thousands of species. Carrot is
a biennial plant, consumed worldwide, whose storage root accumulates high level of carotenoids. The orange carrot was stated
recently until the sixteenth century, being the first emerged domesticated carrot root yellow and purple [1, 2]. The production of
carrots has quadrupled in the last four decades, principally due to
the consumption as fresh vegetable and other high-value products
[3]. Carrot storage root is rich in dietary fiber, antioxidants, and
other nutrients [2], but the most important nutritional value of
carrots lies in the high quantities of α- and β-carotene, making
carrots the richest source of provitamin A in the diet [4].
Manuel Rodrı ´guez-Concepcio ´ n and Ralf Welsch (eds.), Plant and Food Carotenoids: Methods and Protocols,
Methods in Molecular Biology, vol. 2083, https://doi.org/10.1007/978-1-4939-9952-1_24,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
313
Agrobacterium tumefaciens-Mediated Stable
Transformation of Daucus carota
Christian Gonzalez-Calquin and Claudia Stange
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.
Key words Carrot, Transformation protocol, 2,4-dichlorophenoxiacetic acid, Somatic embryogenesis, Agrobacterium tumefaciens, In vitro culture
1 Introduction
Carrot (Daucus carota L., 2n ¼ 18) belongs to the Umbelliferae
group together with parsley (Petroselinum hortense), anise (Pimpinella anisum), celery (Apium graveolens), caraway (Carum carvi),
dill (Anethum graveolens), and other thousands of species. Carrot is
a biennial plant, consumed worldwide, whose storage root accumulates high level of carotenoids. The orange carrot was stated
recently until the sixteenth century, being the first emerged domesticated carrot root yellow and purple [1, 2]. The production of
carrots has quadrupled in the last four decades, principally due to
the consumption as fresh vegetable and other high-value products
[3]. Carrot storage root is rich in dietary fiber, antioxidants, and
other nutrients [2], but the most important nutritional value of
carrots lies in the high quantities of α- and β-carotene, making
carrots the richest source of provitamin A in the diet [4].
Manuel Rodrı ´guez-Concepcio ´ n and Ralf Welsch (eds.), Plant and Food Carotenoids: Methods and Protocols,
Methods in Molecular Biology, vol. 2083, https://doi.org/10.1007/978-1-4939-9952-1_24,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
313