| 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)
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| 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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