Diagnosis of Some Plant Pathogenic Fungi by PCR-ITS Regions

Abstract

This study aimed to diagnosis of plant pathogenic fungi by molecular technique Polymerase Chain Reaction-Internal Transcript Spacer (PCR-ITS). Seven plant pathogenic fungi isolates were isolates from infected Solanaceae plants and diagnosed depending on morphological characteristics. The pathogenicity test was performed for Seven of plant pathogenic fungi isolates. Genomic DNA was extracted from seven plant pathogenic fungi by using Wizard Genomic DNA Purification kit. ITS gene was amplified by using PCR-ITS. The results of the pathogenicity test showed that Macrophomina phaseolina, and Rhizoctonia solani isolates high significance pathogenicity more than other plant pathogenic fungi Fusarium1,Fusarium 2, Fusarium 3,Sclerotinia,andCurvularia which recorded 100%, 96.67%, 86.67%, 70%, 60%, 60%, and 56.67% respectively. The sequencing results of the amplified product of ITS gene using the ITS1 and ITS4 primers obtained were compared with NCBI GenBank by using BLAST analysis. The results of diagnosed depending on morphological characteristics revealed five genera of pathogenic fungi (Fusarium, Curvularia, Sclerotinia, Rhizoctonia and Macrophomina) and also the results of molecular diagnosis confirmed the genus and the species of plant pathogenic fungi as Rhizoctonia solani, Curvularia spicifera, Macrophomina phaseolina, Fusarium oxysporum, Sclerotinia sclerotiorum, Fusarium oxysporum f.sp. cumini and Fusarium oxysporum .