A Diagnostic TaqMan Real-Time PCR Assay for In Planta Detection and Quantification of , Causal Agent of Boxwood Dieback

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Boxwood dieback, caused by , is spreading at an alarming rate in the boxwood industry in the United States. Although . has been accepted as a distinct species within the . species complex, it is difficult to distinguish it from other closely related species based on morphology. Moreover, molecular identification of . requires amplification and sequencing of multiple loci, which can be expensive and time consuming. Therefore, a diagnostic TaqMan real-time PCR assay was developed for early and accurate detection and quantification of . in boxwood. The study involved the design of species-specific primers and a TaqMan probe to differentiate . from other closely related species. The primers and probe discriminate between . and other species in the . species complex and can detect . at very low concentrations, illustrating the high specificity and sensitivity of the assay. This TaqMan real-time PCR assay accurately and rapidly distinguishes boxwood dieback from other diseases with similar symptomatology, including Macrophoma blight, Phytophthora root rot, and Volutella blight, as well as some disorders produced by abiotic agents.

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Plant disease

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