Use of Arabidopsis eceriferum Mutants to Explore Plant Cuticle Biosynthesis

Summary

The plant cuticle is a waxy outer covering on plants that has a primary role in water conservation but is also an important barrier against the entry of pathogenic microorganisms. In this video, we demonstrate the analysis of plant cuticle mutants identified by forward and reverse genetics approaches.

Abstract

The plant cuticle is a waxy outer covering on plants that has a primary role in water conservation, but is also an important barrier against the entry of pathogenic microorganisms. The cuticle is made up of a tough crosslinked polymer called "cutin" and a protective wax layer that seals the plant surface. The waxy layer of the cuticle is obvious on many plants, appearing as a shiny film on the ivy leaf or as a dusty outer covering on the surface of a grape or a cabbage leaf thanks to light scattering crystals present in the wax. Because the cuticle is an essential adaptation of plants to a terrestrial environment, understanding the genes involved in plant cuticle formation has applications in both agriculture and forestry. Today, we'll show the analysis of plant cuticle mutants identified by forward and reverse genetics approaches.

Protocol

I. Visual Screens and Reverse Genetic Approaches to Identify Cuticle Mutants.

Cuticle mutants in the model plant Arabidopsis thaliana are called "eceriferum" mutants, which means "not bearing wax". They can be in visual screens, that is, just by looking at the mutant, you can see that the inflorescence stem of wild-type plants is whitish while the mutants have dark green shiny stems.  One of the mutants that was isolated in this type of visual screen is this one, called eceriferum4 or cer4. 

Another way of discovering interesting cuticle mutants is with a reverse genetics approach: choosing candidate genes of interest a....