Abstract
The pathogens of the Diaporthe genus are now considered to be the dominant pathogens of kiwifruit soft rot. Here, we present a protocol to construct nanoprobes for the Diaporthe genus and detect changes of surface-enhanced Raman spectroscopy for samples collected from infected kiwifruit. We describe steps for synthesis of gold nanoparticles, DNA extraction from kiwifruit, and construction of nanoprobes. We then detail classification of nanoparticles with different aggregation states through dark-field microscope (DFM) picture analysis using Fiji-ImageJ software. For complete details on the use and execution of this protocol, please refer to Yu et al. (2022).(1).