Specific PCR primer designed from genome data for rapid detection of Fusarium oxysporum f. sp. cubense tropical race 4 in the Cavendish banana

Abstract

Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc) tropical race 4 (TR4) severely affects banana production worldwide. Thus, specific PCR primers have been developed to rapidly diagnose and monitor Foc TR4-related fusarium wilt outbreaks in bananas. However, evaluation of these primers revealed room for improvement in the accuracy. This study aimed to design highly specific PCR primers based on genome data for Foc TR4 downloaded from the National Center for Biotechnology Information database. The specificity of the primers was assessed using Foc TR4, Foc races 1 and 2, and 15 other formae speciales strains. The utility of the primers was verified by correctly detecting Foc TR4 in 7 out of 86 isolates of Fusarium spp. obtained from banana farms in the Philippines. The primers allowed for rapid detection in experimentally diseased tissues. We concluded that this novel primer set enables the simplified diagnosis of fusarium wilt caused by Foc TR4 in bananas.