Functional analysis of candidate effector proteins during Sporisorium scitamineum x sugarcane interaction

Abstract

Sugarcane smut is a worldwide distributed disease important to agribusiness, since it can affect cane yield drastically. The disease is caused by the Basidiomycete fungus Sporisorium scitamineum, a biotrophic species that colonizes particularly sugarcane. Cane-smut interaction has been studied by our research group for the past few years in their various aspects, considering both attack of the pathogen and plant defenses. The aim of this project is to study candidate effector proteins of this pathosystem. Effectors are essential to modulate host metabolism to allow pathogen colonization. The identification of such proteins may assist in the recognition of resistance genes relevant to genetic breeding programs. Based on the complete genome sequence of S. scitamineum, candidate genes will be selected in silico (TANIGUTI et al., 2015). Selection strategies will be based on the translated amino acid sequence features, differential expression level of the genes in planta, previously obtained in RNAseq assay (SCHAKER et al., submitted), and genetic diversity studies (BENEVENUTO et al., submitted). Candidate effectors will be analyzed regarding their subcellular location in plant cells using agroinfiltration for transient expression in Nicotiana benthamiana plants. Subsequently, a co-immunoprecipitation assay will be performed to capture protein complexes and evaluate their direct or indirect interaction. The selected candidate genes will have their expression level validated by RT-qPCR. These results will enable future researches considering virulence level of different isolates and also help decision making in plant breeding programs.