Chen Han, Mao Jiana, Fang Yujie, Raza Waqas, Li Zhi, Zhang Chongyuan, Zhu Yingguang, Wang Yuanchao, Dong Suomeng
State Key Laboratory of Agricultural and Forestry Biosecurity, College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China.
Jiangsu Key Laboratory of Pesticide Science and Department of Chemistry, College of Sciences, Nanjing Agricultural University, Nanjing, 210095, China.
Plant Methods. 2025 May 7;21(1):57. doi: 10.1186/s13007-025-01374-9.
Fungicides play an indispensable role in ensuring food security. However, rational chemical selection and fungicide precision application guidance remain constrained by the limitations in real-time monitoring of tracking pathogens within plant tissues. In the current study, we generated a genetically stable Phytophthora infestans strain (PiLuc) expressing luciferase gene, which serves as a dual-mode quantification platform for both in vitro and in vivo throughput screening. Consequently, we designed a 96-well plate high-throughput screening system to assess compounds inhibitory efficacy using PiLuc. Crucially, bioluminescence imaging enabled visualization of PiLuc in potato leaves and tubers during early infection stage, which is invisible to the naked eye. Capitalizing on the semi non-destructive and visual advantages, we developed a system for fungicide bioavailability evaluation and dosage-response assessment in tuber tissues, integrating real-time dynamic monitoring of pathogen. The development of bioluminescent imaging of late blight pathogen establishes an enabling platform for high-throughput fungicide screening while improving the precision bioavailability assessments.
杀菌剂在确保粮食安全方面发挥着不可或缺的作用。然而，合理的化学药剂选择和杀菌剂精准施用指导仍受到植物组织内病原体实时监测跟踪局限性的制约。在本研究中，我们构建了一个表达荧光素酶基因的遗传稳定的致病疫霉菌株（PiLuc），它作为一个用于体外和体内高通量筛选的双模式定量平台。因此，我们设计了一个96孔板高通量筛选系统，使用PiLuc评估化合物的抑制效果。至关重要的是，生物发光成像能够在早期感染阶段可视化马铃薯叶片和块茎中的PiLuc，这是肉眼不可见的。利用半非破坏性和可视化优势，我们开发了一个用于块茎组织中杀菌剂生物利用度评估和剂量反应评估的系统，整合了病原体的实时动态监测。晚疫病病原体生物发光成像的发展建立了一个用于高通量杀菌剂筛选的平台，同时提高了生物利用度评估的精度。
