Quantitative High-Throughput, Real-Time Bioassay for Plant Pathogen Growth in vivo

Abstract

Effective assessment of pathogen growth can facilitate screening for disease resistance, mapping of resistance loci, testing efficacy of control measures, or elucidation of fundamental host-pathogen interactions. Current methods are often limited by subjective assessments, inability to detect pathogen growth prior to appearance of symptoms, destructive sampling, or limited capacity for replication and quantitative analysis. In this work we sought to develop a real-time, in vivo , high-throughput assay that would allow for quantification of pathogen growth. To establish such a system, we worked with the broad host-range, highly destructive, soil-borne oomycete pathogen, Phytophthora capsici . We used an isolate expressing red fluorescence protein (RFP) to establish a microtiter plate, real-time assay to quantify pathogen growth in live tissue. The system was successfully used to monitor P. capsici growth in planta on cucumber ( Cucumis sativus ) fruit and pepper ( Capsicum annuum ) leaf samples in relation to different levels of host susceptibility. These results demonstrate usefulness of the method in different species and tissue types, allowing for highly replicated, quantitative time-course measurements of pathogen growth in vivo . Analyses of pathogen growth during initial stages of infection preceding symptom development show the importance of very early stages of infection in determining disease outcome, and provide insight into points of inhibition of pathogen growth in different resistance systems.

Plant phenotyping relevance

植物組織上の病原体増殖を蛍光でリアルタイム・定量測定する高スループット手法を開発し、複数の植物種・組織で検証している。病原体検出のみでなく、生体植物の感染進行・病害状態に関わる表現型取得が中心である。

Code and data availability

The article describes a fluorescence-based microplate bioassay for Phytophthora capsici growth, but no public phenotype dataset, image/sensor data, author analysis code, or trained model is deposited. The only supplementary link points to supplementary figures (fluorescence curves and Gompertz model figures), not raw数据

No evidence-backed public reproduction asset is currently recorded.