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Culture‐Free Detection of Crop Pathogens at the Single‐Cell Level by Micro‐Raman Spectroscopy
Advanced Science ( IF 14.3 ) Pub Date : 2017-07-10 , DOI: 10.1002/advs.201700127 Qinhua Gan 1, 2 , Xuetao Wang 3 , Yun Wang 4 , Zhenyu Xie 1 , Yang Tian 5 , Yandu Lu 1, 6
Advanced Science ( IF 14.3 ) Pub Date : 2017-07-10 , DOI: 10.1002/advs.201700127 Qinhua Gan 1, 2 , Xuetao Wang 3 , Yun Wang 4 , Zhenyu Xie 1 , Yang Tian 5 , Yandu Lu 1, 6
Affiliation
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The rapid and sensitive identification of invasive plant pathogens has important applications in biotechnology, plant quarantine, and food security. Current methods are far too time‐consuming and need a pre‐enrichment period ranging from hours to days. Here, a micro‐Raman spectroscopy‐based bioassay for culture‐free pathogen quarantine inspection at the single cell level within 40 min is presented. The application of this approach can readily and specifically detect plant pathogens Burkholderia gladioli pv. alliicola and Erwinia chrysanthemi that are closely related pathogenically. Furthermore, the single‐bacterium detection was able to discriminate them from a reference Raman spectral library including multiple quarantine‐relevant pathogens with broad host ranges and an array of pathogenic variants. To show the usefulness of this assay, Burkholderia gladioli pv. alliicola and Erwinia chrysanthemi are detected at single‐bacterium level in plant tissue lesions without pre‐enrichment. The results are confirmed by the plate‐counting method and a genetic molecular approach, which display comparable recognition ratios to the Raman spectroscopy‐based bioassay. The results represent a critical step toward the use of micro‐Raman spectroscopy in rapid and culture‐free discrimination of quarantine relevant plant pathogens.
中文翻译：
显微拉曼光谱法在单细胞水平上无培养物检测农作物病原体
快速而敏感地识别入侵性植物病原体在生物技术，植物检疫和食品安全中具有重要的应用。当前的方法过于费时，并且需要一个数小时到数天的预富集期。本文介绍了一种基于显微拉曼光谱的生物测定法，可在40分钟内在单个细胞水平上对无培养物的病原体进行检疫检查。该方法的应用可以轻松，特异性地检测植物病原体伯克霍尔德菌。阿利科拉和欧文氏菊与病因密切相关 此外，单细菌检测能够将它们与参考拉曼光谱库区分开来，该库包括多个与检疫相关的病原体，具有广泛的宿主范围和一系列致病变体。为了显示该测定法的有用性，伯克霍尔德氏菌pv。阿利科拉和欧文氏菊无需预先富集即可在植物组织病变中以单细菌水平检测到。结果通过板计数法和遗传分子方法得到证实，与基于拉曼光谱的生物测定法相比，它们显示出可比的识别率。结果表明，在快速和无培养的检疫相关植物病原体鉴别中，使用微拉曼光谱法迈出了关键的一步。
更新日期：2017-07-10
中文翻译：
显微拉曼光谱法在单细胞水平上无培养物检测农作物病原体
快速而敏感地识别入侵性植物病原体在生物技术，植物检疫和食品安全中具有重要的应用。当前的方法过于费时，并且需要一个数小时到数天的预富集期。本文介绍了一种基于显微拉曼光谱的生物测定法，可在40分钟内在单个细胞水平上对无培养物的病原体进行检疫检查。该方法的应用可以轻松，特异性地检测植物病原体伯克霍尔德菌。阿利科拉和欧文氏菊与病因密切相关 此外，单细菌检测能够将它们与参考拉曼光谱库区分开来，该库包括多个与检疫相关的病原体，具有广泛的宿主范围和一系列致病变体。为了显示该测定法的有用性，伯克霍尔德氏菌pv。阿利科拉和欧文氏菊无需预先富集即可在植物组织病变中以单细菌水平检测到。结果通过板计数法和遗传分子方法得到证实，与基于拉曼光谱的生物测定法相比，它们显示出可比的识别率。结果表明，在快速和无培养的检疫相关植物病原体鉴别中，使用微拉曼光谱法迈出了关键的一步。
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