拉曼光谱技术在植物病原菌检验检疫中的应用

外来有害物种，尤其是植物病原菌的传播风险巨大。传统理化手段在植物病原菌检疫领域面临着诸多挑战：周期长、假阴性高、灵敏度低等。分子生物学技术一定程度上提高了灵敏度和准确度。但其需要繁琐的菌种培养和基因组提取过程，耗时耗力，且一菌一标准，广谱性不强。因此，迫切需要一种快速高效广谱的病原菌检疫方法。拉曼光谱分析技术具有原位、快速、广谱性强的优势，在植物检验检疫领域具有巨大的应用潜力，却少有应用报道。本项目将：（1）以15种重要植物病原菌为模式，解析其拉曼图谱，构建“拉曼指纹库I”；（2）结合分子生物学方法和BIOLOG技术构建重要未知病原细菌的“拉曼指纹库II”；（3）基于库I和库II，对比健康植株与带病植株病变组织拉曼图谱，探索该技术在植物组织原位检测中的应用。本研究利用拉曼光谱技术实现病原菌的无创、原位、快速、灵敏的检测，将有效解决检疫事业的发展瓶颈，推动检疫工作的发展。

The invasive exotic harmful organisms, particularly the phytopathogen is highly risked. Tranditional biochemical strategies encounter innate defects: time-counsuming, more false negative, low sensitivity et al. Molecular technologies improve the sensitivity and accurary to some degree, however, it suffers from the time and labor costing process for cell culture and genomic DNA extraction. In addition, the high selectivity of each molecular standard limits their applicability. Therefore, a rapid, efficient and broad-spectrum phytopathogen inspection approach is needed. Raman spectroscopy is in situ, fast, broad-spectrum, and has great potential for application in the field of plant inspection and quarantine, but yet few application has been documented. Therefore, in this proposal, a novel Raman spectroscopy-assisted phytopathogen inspection and quarantine technology is presented. i) construct ‘Raman fingerprint database I’ after collection and illumination of the Raman spectrum of 15 important quarantine plant pathogens; ii) construct ‘Raman fingerprint database II’ of unknown phytopathogen combined with the molecular and biochemical methods; iii) compare the Raman spectrums of tissues from health and disease plants to verify the feasibility of in situ application of Raman technology. The proposed work provide an invasive, in situ, rapid and sensitive quarantine technolgy which offer an opportunity to solve the limits of tranditional quarantine technolgy and thereby create unprecedented possibilities to consummate the quarantine research.