Development of specific primers based on the genomes of Penicillium spp. for rapid detection of Penicillium digitatum among fungal isolates in citrus

Specific primers targeting Penicillium digitatum were developed based on fungal genes RPB1 and cmd, which are conserved among the genomes of Penicillium spp. The specific primers were designed based on the mutational sites in the homologous regions of the conserved genes. The results indicated that primer pairs RPB1-1 and cmd-3 were specific enough to distinguish Penicillium digitatum (N1) from Penicillium chrysogenum (Q), Penicillium italicum (A10) and Penicillium expansum (L) when the DNA samples were diluted 100-fold. To further verify the effectiveness and specificity of the two primer pairs RPB1-1 and cmd-3, 38 strains of fungal isolates from sources related to citrus were detected using both primer pairs, and 14 candidate P. digitatum strains were identified. Then, the fourteen candidate P. digitatum strains were further identified as P. digitatum by morphological and molecular methods, which confirmed the detection accuracy and reliability of the specific primer pairs RPB1-1 and cmd-3 as molecular markers of P. digitatum. This work may significantly facilitate the rapid identification of P. digitatum in the citrus industry.