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Furuta, Kazuyoshi,Nagashima, Saki,Inukai, Tsuyoshi,Masuta, Chikara The Korean Society of Plant Pathology 2017 Plant Pathology Journal Vol.33 No.1
One of the major problems in strawberry production is difficulty in diagnosis of anthracnose caused by Colletotrichum acutatum or Glomerella cingulata in latent infection stage. We here developed a diagnostic tool for the latent infection consisting of initial culturing of fungi, DNA extraction, synthesis of PCR-amplified probes and microtube hybridization (MTH) using a macroarray. The initial culturing step is convenient to lure the fungi out of the plant tissues, and to extract PCR-inhibitor-free DNA directly from fungal hyphae. For specific detection of the fungi, PCR primers were designed to amplify the fungal MAT1-2 gene. The subsequent MTH step using the PCR products as probes can replace the laborious electrophoresis step providing us sequence information and high-throughput screening. Using this method, we have conducted a survey for a few thousands nursery plants every year for three consecutive years, and finally succeeded in eliminating latent infection in the third year of challenge.
Kazuyoshi Furuta,Saki Nagashima,Tsuyoshi Inukai,Chikara Masuta 한국식물병리학회 2017 Plant Pathology Journal Vol.33 No.1
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.