chapter  thirty-two
16 Pages

Transformation of tobacco and carrot using Agrobacterium tumefaciens and expression of the β-glucuronidase (GUS) reporter gene

ByIgnacio E. Maldonado-Mendoza, Melina López-Meyer, Craig L. Nessler

This chapter uses the reporter gene system to study gene expression in transgenic plants. The wide acceptance of β-glucuronidase gene (GUS) as a reporter gene is mainly due to the fact that it can be quickly assayed without using radioactive substrates. The chapter discusses the Agrobacterium tumefaciens-based transformation. It examines the use of reporter and marker genes as a way to confirm plant transformation. The chapter then explores the expression of a gene under the regulation of an inducible and a constitutive promoter and provides different patterns of plant gene expression. Tobacco contains nicotine, which is a health-threatening, addictive drug. Tobacco provides one of the most easy-to-use systems for plant transformation because of its high transformation and regeneration frequencies and relatively short life cycle. Carrot is an edible biennial plant which produces a modified, fleshy storage root. Somatic embryogenesis in carrot has been used as a model system for the study of plant development for over thirty years.