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Foot rot disease caused by  Diaporthe destruens  (formerly  Plenodomus destruens ) has become a major concern for the production of sweet potato [ Ipomoea batatas  (L.) Lam.] in Japan. A related fungus  Diaporthe batatas , which causes dry rot disease of sweet potato, is native and is widespread in fields in Japan. The similar characteristics of these two pathogens pose a challenge for conventional disease diagnosis. Currently, there are no effective molecular measures for identifying and distinguishing  D. destruens  and  D. batatas . Here, we demonstrate a real-time PCR assay that distinguishes and quantifies  D. batatas  and  D. destruens  from co-infected sweet potato. The assay was performed with various simulated DNA combinations of  D. batatas  and  D. destruens  ranging from 1:1 to 1:1. The assay was also used with the ratios of  D. batatas :  D. destruens : sweet potato DNA ranging from 1:1:1 to 1:1:1. These assays produced a specific amplification product for each of the pathogens, and quantified the fungal biomass over the entire range tested without detecting false positives. The assay was validated by using infected sweet potato collected from various fields; it showed sufficient sensitivity and specificity to quantify and distinguish  D. batatas  and  D. destruens  from these field samples. Thus, our real-time PCR assay would be a useful tool for diagnosis of  D. batatas  and  D. destruens  and is expected to provide the foundation for the design of integrated disease management strategies for foot rot disease in sweet potato.

Real-Time PCR Assay for the Diagnosis and Quantification of Co-infections by Diaporthe batatas and Diaporthe destruens in Sweet Potato