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STAR Protocols · 2023 · IF 0

Protocol for early warning of Diaporthe infection in kiwifruit soft rot by plasmonic dimer-enhanced Raman spectroscopy

Hao Jiang,Ting Yu,Min Miao,Honglin Liu

Summary 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

研究领域: 植物病理学 · 分子生物学 · 光谱学

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iScience · 2022 · IF 6.107

Early warning of Diaporthe infection in kiwifruit soft rot by plasmonic dimer-enhanced Raman spectroscopy

Ting Yu,Huanhuan Li,Baomei Zhou,Kui Liu,Hao Jiang,Min Xu,Mengke Su,Yongsheng Liu,Min Miao,Honglin Liu

Summary Diaporthe genus is the dominant pathogens of kiwifruit soft rot with long incubation period and rapid onset and very hard to detect in advance. It is of great significance to develop point-of-care tests for disease prevention and field management. Here we screen a pair of genus-level primers for constructing plasmonic dimer Rayleigh/Raman spectroscopy, which enables rapid, specific, and sensitive detection of Diaporthe genus in real kiwifruits. Dark-filed Rayleigh scattering clearly visualizes the target-induced dimer assembly of nanoprobes. Plasmonic dimer-enhanced Raman spectroscopy has high specificity for 12 common isolates of Diaporthe genus in one batch samples with highly accurate results and much higher sensitivity than polymerase chain reaction (PCR). It realizes the recognition of Diaporthe infection at an early stage of 24 h when the kiwifruits do not have any noticeable symptoms. It demonstrates a bright prospect of point-of-care tests for early warning of kiwifruit soft rot infection and quality control.

研究领域: 农业 · 植物病理学 · 光谱学

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Protocol for early warning of Diaporthe infection in kiwifruit soft rot by plasmonic dimer-enhanced Raman spectroscopy

Early warning of Diaporthe infection in kiwifruit soft rot by plasmonic dimer-enhanced Raman spectroscopy