Chemical screening pipeline for identification of specific plant autophagy modulators

Abstract

Autophagy is a major catabolic process in eukaryotes and is implicated in responses to biotic and abiotic stresses, programs of aging and cell death. Autophagy is known to regulate agriculturally important traits such as plant fitness, longevity, biomass and seed yield. However, despite its obvious significance, there is still a shortage of reliable tools modulating plant autophagy. Here we describe the first robust pipeline for identification of specific plant autophagy-modulating compounds. Our novel screening protocol comprises four phases: (i) high-throughput screening of chemical compounds in cell cultures of Nicotiana tabacum; (ii) confirmation of the identified hits in planta using Arabidopsis thaliana; (iii) further characterization of the effect using conventional molecular biology methods; (iv) verification of specificity of the effect to autophagy in planta. The methods detailed here streamline identification of specific plant autophagy modulators and greatly aid in unravelling the molecular mechanisms of plant autophagy.