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.
One of the major problems in strawberry production is difficulty in diagnosis of anthracnose caused byColletotrichum acutatumorGlomerella cingulatain 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 fungalMAT1-2gene. 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.
Agroscience Research Laboratories, Plant Biotechnology Center, Hokusan Co., Ltd., Kitahiroshima 061-1111, Japan.
Kazuyoshi
K
Furuta
Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
Saki
S
Nagashima
Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
Tsuyoshi
T
Inukai
Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
Chikara
C
Masuta
Colletotrichum; microtube-hybridization; strawberry