Construction of a System for the Strawberry Nursery Production towards Elimination of Latent Infection of Anthracnose Fungi by a Combination of PCR and Microtube Hybridization

다국어 초록 (Multilingual Abstract)

One of the major problems in strawberry productionis difficulty in diagnosis of anthracnose caused by Colletotrichumacutatum or Glomerella cingulata in latentinfection stage. We here developed a diagnostic toolfor the latent infection consisting of initial culturingof fungi, DNA extraction, synthesis of PCR-amplifiedprobes and microtube hybridization (MTH) using amacroarray. The initial culturing step is convenient tolure the fungi out of the plant tissues, and to extractPCR-inhibitor-free DNA directly from fungal hyphae.
For specific detection of the fungi, PCR primers weredesigned to amplify the fungal MAT1-2 gene. The subsequentMTH step using the PCR products as probescan replace the laborious electrophoresis step providingus sequence information and high-throughputscreening. Using this method, we have conducted asurvey for a few thousands nursery plants every yearfor three consecutive years, and finally succeeded ineliminating latent infection in the third year of challenge.

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