Identification and inference of the evolutionary relationship of seven known Ribonucleic acid (RNA) interference components Fusarium circinatum, its closely related species as well as other fungi outside of the genus. Twenty-one Fusarium species and thirteen species from other genera were chosen. The genes of interest include Argonaute, Dicer, RNA-dependent RNA polymerase, Replication protein A1 and A2, Mitochondrial ribosomal protein L3 (MRPL3) and Quelling Deficient-2 interating protein (QIP). A proof of concept of the functionality of RNA interference (RNAi) in Fusarium circinatum. These was done by applying double-stranded RNA targeting a fluorescence gene in Fusarium circinatum.