title: Species-Specific Detection of Mycosphaerella polygoni-cuspidati as a Biological Control Agent for Fallopia japonica by PCR Assay.
abstract: The ascomycete fungus Mycosphaerella polygoni-cuspidati has been undergoing evaluation as a potential classical biological control agent for the invasive weed Fallopia japonica (Japanese knotweed), which has become troublesome in Europe and North America. In advance of the potential release of a biocontrol agent into a new environment, it is crucial to develop an effective monitoring system to enable the evaluation of agent establishment and dispersal within the target host population, as well as any potential attacks on non-target species. Therefore, a primer pair was designed for direct, rapid, and specific detection of the Japanese knotweed pathogen M. polygoni-cuspidati based on the sequences of the internal transcribed spacer regions including the 5.8S rDNA. A PCR product of approximately 298 bp was obtained only when the DNA extracted from mycelial fragments of M. polygoni-cuspidati was used. The lower limit of detection of the PCR method was 100 fg of genomic DNA. Using the specific primer pair, M. polygoni-cuspidati could be detected from both naturally and artificially infected Japanese knotweed plants. No amplification was observed for other Mycosphaerella spp. or fungal endophytes isolated from F. japonica. The designed primer pair is thus effective for the specific detection of M. polygoni-cuspidati in planta.
authors.affiliation: CABI Europe-UK, Bakeham Lane, Egham, Surrey, TW20 9TY, UK. D.Kurose@cabi.org.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kurose
authors.affiliation: Laboratory of Plant Pathology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan.
authors.forename: Naruto
authors.initials: N
authors.lastname: Furuya
authors.affiliation: Laboratory of Plant Pathology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Saeki
authors.affiliation: Laboratory of Plant Pathology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan.
authors.forename: Kenichi
authors.initials: K
authors.lastname: Tsuchiya
authors.affiliation: Plant Pathology Laboratory, Faculty of Agriculture, Tokyo University of Agriculture, 1737 Funako, Atsugi, Kanagawa, 243-0034, Japan.
authors.forename: Seiya
authors.initials: S
authors.lastname: Tsushima
authors.affiliation: CABI Europe-UK, Bakeham Lane, Egham, Surrey, TW20 9TY, UK.
authors.forename: Marion K
authors.initials: MK
authors.lastname: Seier
mesh_terms: D001203:Ascomycota / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D061046:Biological Control Agents / Q000032:analysis / Q000494:pharmacology; D017931:DNA Primers / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D045468:Fallopia japonica / Q000382:microbiology*; D057230:Limit of Detection; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity
chemical_list: D061046:Biological Control Agents; D017931:DNA Primers; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer
keywords: Japanese knotweed; Leaf spot disease; Molecular marker; Weed biocontrol; rDNA-ITS region