Establishment of ultra-rapid identification of spider mite species based on RNA-sequencing
Project/Area Number |
17H03775
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Research Category |
Grant-in-Aid for Scientific Research (B)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Plant protection science
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Research Institution | Ryutsu Keizai University (2018-2020) Ibaraki University (2017) |
Principal Investigator |
Gotoh Tetsuo 流通経済大学, 経済学部, 教授 (60178449)
|
Co-Investigator(Kenkyū-buntansha) |
北嶋 康樹 茨城大学, 農学部, 准教授 (00431651)
坂本 洋典 国立研究開発法人国立環境研究所, 生物・生態系環境研究センター, 研究員 (70573624)
粥川 琢巳 国立研究開発法人農業・食品産業技術総合研究機構, 生物機能利用研究部門, 主任研究員 (70580463)
|
Project Period (FY) |
2017-04-01 – 2021-03-31
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Project Status |
Completed (Fiscal Year 2020)
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Budget Amount *help |
¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000)
Fiscal Year 2020: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2019: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2018: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2017: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
|
Keywords | ダニ・線虫管理 / 系統関係 / ゲノム |
Outline of Final Research Achievements |
Spider mite species are extremely difficult to identify only by morphology. In this study, a molecular method was established for ultra-rapid identification of spider mites: (1) Species-specific primer sequences were extracted by comprehensive analysis of RNA-Seq data. (2) PCR was performed using the candidate primers to confirm species-specific amplification. For multiplex PCR to identify species, primer sets were designed resulting in PCR amplification products differing among species. (3) Multiplex PCR conditions were examined using mixtures of primer sets, and it was confirmed that up to six species can be identified with one PCR tube. (4) To speed up DNA extraction, we arranged PCR conditions using either an individual mite directly or a mixture of homogenized mite body as template. Finally, we tested our method to correctly identify 12 strains of 11 spider mite species within 1.5 h using multiplex PCR.
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Academic Significance and Societal Importance of the Research Achievements |
多数の農業害虫を含むハダニにおける害虫種の簡易識別は、農業や検疫の現場で広く望まれる。本研究の成果は、特別な機器を必要とせず、PCR法の応用により、1.5時間という短時間で害虫種のハダニの識別に成功し、普及しやすい識別技術として社会の要請に応えるものである。また本研究では、RNA-Seq(全転写産物解析)により得られた大規模データから種特異的配列を得る手法を開発し、学術的にも大きく貢献した。
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Report
(5 results)
Research Products
(15 results)