Agroinfiltration for Enhanced Transgene Expression in Coffee Leaves (Coffea arabica L.)

Methods Mol Biol. 2024:2788:227-241. doi: 10.1007/978-1-0716-3782-1_13.

Abstract

The Coffea spp. plant is a significant crop in Latin America, Africa, and Asia, and recent advances in genomics and transcriptomics have opened possibilities for studying candidate genes and introducing new desirable traits through genetic engineering. While stable transformation of coffee plants has been reported using various techniques, it is a time-consuming and laborious process. To overcome this, transient transformation methods have been developed, which avoid the limitations of stable transformation. This chapter describes an ex vitro protocol for transient expression using A. tumefaciens-mediated infiltration of coffee leaves, which could be used to produce coffee plants expressing desirable traits against biotic and abiotic stresses, genes controlling biochemical and physiological traits, as well as for gene editing through CRISPR/Cas9.

Keywords: Agrobacterium-mediated transformation; Gene function analysis; Genetic engineering; Leaf infiltration; Transgene expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium tumefaciens* / genetics
  • CRISPR-Cas Systems
  • Coffea* / genetics
  • Gene Editing* / methods
  • Gene Expression Regulation, Plant
  • Plant Leaves* / genetics
  • Plant Leaves* / metabolism
  • Plants, Genetically Modified* / genetics
  • Transformation, Genetic
  • Transgenes*