Construction of pyrosequencing-based assays to determine frequency for fungicide-resistance mutation alleles in genomic DNA pools

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K07629
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Plant protection science
• Research Institution: Toyo University
• Principal Investigator: Fujimura Makoto 東洋大学, 生命科学部, 教授 (50297735)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: パイロシークエンス法 / サスペンションアレイ法 / キュウリ褐斑病菌 / 灰色かび病菌 / 殺菌剤耐性

Outline of Final Research Achievements

Many specific-MOA-fungicides have been developed and contributed to crop and vegetable production though disease control, however, the problem of fungicide resistance becomes serious and more complicated in many fungal plant pathogens particularly of cucumber and tomato. In this study, we constructed the pyrosequencing assays and the suspension-array assays for the fungicide-resistance mutations in various plant pathogen such as Botrytis cinerea and Corynespora cassiicola. Pyrosequencing assays were useful not only to detect the fungicide resistance mutations in each isolate of plant pathogen, but also to estimate the frequency of fungicide resistance alleles in pooled DNA. In contrast, the suspension-array assays could be used to determine the fungicide resistance mutations in several fungicide-target genes in a single reaction. The combination of these two tools contributes to monitoring and management of the fungicide-resistance pathogens in each field.

Free Research Field

農薬科学

Academic Significance and Societal Importance of the Research Achievements

世界人口の増加にともない、安全で安定した農作物を生産する技術の重要度は増している。病害防除分野では、農業用殺菌剤の耐性菌が出現し、深刻な問題を与えている。本研究は、複雑化する圃場における耐性菌をモニターする2種類の遺伝子診断法を提案するものであり、学術的な意義をもつ。また、耐性菌の動態をモニターできるようになることから、無駄な農薬散布を抑制した減農薬に貢献する。従って、食の安全・安心および環境にやさしい農業に貢献する意義がある。