Culture-Free Detection of Crop Pathogens at the Single-Cell Level by Micro-Raman Spectroscopy.
Culture-Free Detection of Crop Pathogens at the Single-Cell Level by Micro-Raman Spectroscopy.
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DOI：
10.1002/advs.201700127
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发表时间：
2017-11
期刊：
影响因子：
--
通讯作者：
Lu Y
中科院分区：
文献类型：
--
作者：
Gan Q;Wang X;Wang Y;Xie Z;Tian Y;Lu Y
入侵植物病原菌的快速、灵敏鉴定在生物技术、植物检疫和食品安全等方面具有重要应用。目前的方法过于耗时，需要数小时至数天的预富集期。本文介绍了一种基于微拉曼光谱的生物测定法，可在40 min内完成单细胞水平的无培养病原体检疫检验。应用该方法可以快速、特异地检测植物病原菌唐菖蒲伯克霍尔德氏菌。大蒜生菌和欧文氏菌在病原学上关系密切。此外，单细菌检测能够将其与参考拉曼光谱库区分开来，该参考拉曼光谱库包括具有广泛宿主范围和一系列致病性变体的多种检疫相关病原体。为了显示该测定的有用性，唐菖蒲伯克霍尔德氏菌致病变种（Burkholderia gladioli pv.）在没有预富集的情况下，在植物组织病变中以单细菌水平检测到大蒜生菌和欧文氏菌。通过平板计数法和遗传分子方法证实了结果，其显示出与基于拉曼光谱的生物测定相当的识别率。这些结果代表了使用显微拉曼光谱在快速和无培养的检疫相关植物病原体的歧视的关键一步。
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.
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影响因子：
2.7
作者：
He, Y.;Munkvold, G. P.
通讯作者：
Munkvold, G. P.
影响因子：
4.4
作者：
Rösch, P;Harz, M;Popp, J
通讯作者：
Popp, J
影响因子：
14.9
作者：
Chenna, R;Sugawara, H;Thompson, JD
通讯作者：
Thompson, JD
用于单细胞表型分析的凝聚拉曼光谱
影响因子：
3
作者：
Sun S;Wang X;Gao X;Ren L;Su X;Bu D;Ning K
通讯作者：
Ning K
影响因子：
4.3
作者：
Assaf, A.;Cordella, C. B. Y.;Thouand, G.
通讯作者：
Thouand, G.