Species-Specific Detection of Mycosphaerella polygoni-cuspidati as a Biological Control Agent for Fallopia japonica by PCR Assay

Daisuke Kurose, Naruto Furuya, Tetsuya Saeki, Kenichi Tsuchiya, Seiya Tsushima, Marion K. Seier

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The ascomycete fungus Mycosphaerellapolygoni-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.

Original languageEnglish
Pages (from-to)626-633
Number of pages8
JournalMolecular Biotechnology
Volume58
Issue number10
DOIs
Publication statusPublished - Oct 1 2016

Fingerprint

Fallopia japonica
Biological Control Agents
Assays
Biocontrol
Polymerase Chain Reaction
DNA
Pathogens
Ribosomal DNA
Fungi
Amplification
Monitoring
Endophytes
Ascomycota
Health Services Needs and Demand
North America
Limit of Detection

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology

Cite this

Species-Specific Detection of Mycosphaerella polygoni-cuspidati as a Biological Control Agent for Fallopia japonica by PCR Assay. / Kurose, Daisuke; Furuya, Naruto; Saeki, Tetsuya; Tsuchiya, Kenichi; Tsushima, Seiya; Seier, Marion K.

In: Molecular Biotechnology, Vol. 58, No. 10, 01.10.2016, p. 626-633.

Research output: Contribution to journalArticle

Kurose, Daisuke ; Furuya, Naruto ; Saeki, Tetsuya ; Tsuchiya, Kenichi ; Tsushima, Seiya ; Seier, Marion K. / Species-Specific Detection of Mycosphaerella polygoni-cuspidati as a Biological Control Agent for Fallopia japonica by PCR Assay. In: Molecular Biotechnology. 2016 ; Vol. 58, No. 10. pp. 626-633.
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