Abstract
Effective quantification of Calonectria ilicicola in naturally infested soils is critical for evaluating and validating control measures for managing red crown rot disease in soybean. Counting of microsclerotia has been used to quantify this pathogen, but the method is laborious and time-consuming to process large numbers of soil samples using wet sieving. To overcome these issues, we developed and tested a qPCR method to detect and quantify C. ilicicola in naturally infested soil of soybean fields. The primer pairs and probe, designed based on the intergenic spacer region of the ribosomal DNA, showed high specificity with a single melting curve peak in DNA samples from nine C. ilicicola isolates. No amplification product was detected with 2 isolates of other Calonectria species and 12 isolates of common soil-borne fungi. There was a significant positive correlation between the amount of C. ilicicola DNA and the number of C. ilicicola microsclerotia. The qPCR method is a promising tool to quantify C. ilicicola in soil.
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