Chemical Mutagenesis of Embryogenic Cell Suspensions of Coffea arabica L. var. Catuaí Using EMS and NaN3

Abstract

AbstractChemical mutagens, such as ethyl methanesulfonate (EMS) and sodium azide (NaN3), interact with DNA and can primarily induce single base modifications along the genome. Populations derived from chemical mutagenesis experiments are presumed to harbor high density of point mutations in the genome. Therefore, this technique, along with in vitro culture methods such as somatic embryogenesis (SE), can introduce genetic variation in otherwise genetically homogeneous populations. In vitro mutagenesis of embryogenic cell suspension cultures represents an efficient method to quickly develop mutant plantlets of unicellular origin. The development of mutant populations in this important crop represents a fundamental steppingstone in the development of novel varieties and the characterization of candidate genes involved in traits such as disease resistance, grain metabolite content and flowering induction. This chapter describes the protocol for establishment of embryogenic cell suspension cultures as well as methods of mutation induction using EMS and NaN3 on embryogenic cell suspensions of C. arabica, variety Catuaí. Furthermore, this chapter includes a protocol for mutant plant regeneration in in vitro conditions.