岡本 晃充47Akimitsu OKAMOTO高野 義孝48Yoshitaka TAKANO合成化学とゲノム医科学の融合によるメチル化DNA液体生検法の創出（2018採択）Development of methylated DNA-based liquid biopsy through coalition between synthetic chemistry and genome medicinal science(Project 2018)植物の生存を保証する侵入後抵抗性の分子機構解明と応用展開（2018採択）Dissection of the molecular mechanism of postinvasive resistance essential for plant survival and its application and development(Project 2018)45低侵襲な簡便生体分析法が求められており，我々は血中循環DNA（cfDNA）のメチル化レベルの簡便液体生検法を検討した．ここでは，メチル化DNAを配列選択的に捕捉できるICONプローブ修飾ビーズを最新肝臓がん患者メチロームデータをもとに作成するとともに，患者由来血液をセットするだけで目的のメチル化cfDNAを回収できるメチル化DNAオートコレクターを開発した．その結果，肝臓がん患者に由来する微量cfDNAを少量の血中から短時間で検出できるシステムを確立した．In response to the need for a minimally invasive and simple bioanalytical method, we investigated a simple liquid biopsy method to determine the methylation level of circulating DNA (cfDNA) in blood. Here, we de-veloped ICON probe-modified beads for sequence-selective capture of methylated DNA based on the latest methylome data of liver cancer patients, and a methylated DNA auto-collector that can collect the target methylated cfDNA by simply setting the patient-derived blood. As a result, we have established a system that can detect trace amounts of cfDNA derived from the blood of liver cancer patients in a short time.高等植物は，侵入を阻止する抵抗性が突破され，植物病原糸状菌が侵入した場合，「侵入後抵抗性」と呼ばれる第2層目の防御を新たに活性化させ菌の進展を抑止する．したがって，侵入後抵抗性は，高等植物の生存を保証する極めて重要な生体防御機構であるが，その分子機構は不明な点が多い．今回，シロイヌナズナの侵入後抵抗性の分子機構に焦点を当て，さらに，この侵入後抵抗性を活性化し，病害防除効果を示す化合物を探索した．その結果，侵入後抵抗性に関わるシロイヌナズナの新規の因子の発見に成功し，さらにシロイヌナズナの生物発光レポーターラインを作出し，これを利用することで，侵入後抵抗性を活性化し，植物病原糸状菌への防除効果を示す複数の化合物の同定に成功した．When phytopathogenic fungi overcome the defense layer that terminates invasion trial, higher plants newly activate the second layered defense called postinvasive resistance to terminate further expansion of fungal in-vasive hyphae. Therefore, postinvasive resistance is critical defense mechanism that guarantees plant surviv-al, however its molecular mechanism is poorly understood. In this study, we focused on molecular mecha-nism of postinvasive resistance in Arabidopsis thaliana, and also tried to identify chemical compounds that exhibit disease control effects by activating postinvasive resistance. As a result, we identified novel factors involved in postinvasive resistance in A. thaliana. Furthermore, by using bioluminescence reporter lines of A. thaliana, we successfully identified multiple chemicals that have disease control effects via activation of post-invasive resistance.
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