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具有检测海枣中东红棕象甲(Rhynchophorus ferrugineus)能力的全细胞细菌生物传感器:概念验证研究。

Whole-cell bacterial biosensor with the capability to detect red palm weevil, Rhynchophorus ferrugineus, in date palm trees, Phoenix dactylifera: a proof of concept study.

机构信息

Institute of Postharvest and Food Science, Department of Postharvest Science, Volcani Institute, Agricultural Research Organization, Rishon LeZion 7505101, Israel; Institute of Biochemistry, Food Science and Nutrition, Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot 76100, Israel.

Eden Farm, Agricultural R&D center, Emek HaMa'ayanot Regional Council, Beit Shean Valley 171000, Israel.

出版信息

J Biotechnol. 2022 Sep 20;357:47-55. doi: 10.1016/j.jbiotec.2022.08.007. Epub 2022 Aug 11.

DOI:10.1016/j.jbiotec.2022.08.007
PMID:35963593
Abstract

The red palm weevil (RPW), Rhynchophorus ferrugineus, is considered a severe pest of palms. Usually, the early stages of infection are without visible signs. An attractive early sensing approach of non-visible infections is based on volatile organic compounds (VOCs). In this study, a whole-cell bacterial biosensor was used for the identification of RPW in date palm (Phoenix dactylifera). The cells are genetically modified to produce light in the presence of general stresses. The bioluminescent bacterial panel is based on three genetically engineered Escherichia coli strains that are sensitive to cytotoxicity (TV1061), genotoxicity (DPD2794), or quorum-sensing (K802NR). The bioluminescent bacterial panel detects the presence of VOCs and a change in the light signal is then generated, reflecting the health status of the date palm tree. The bioreporter bacteria cells are immobilized in calcium alginate tablets and placed in a sealed jar without direct contact with the tested sample, thereby exposing them only to the VOCs in the surrounding air. The immobilized bacteria cells were exposed to the air near infected by RPW or uninfected sugar canes, date palm tree pieces, and on date palm trees. Commercial plate reader was used for signal measurement. The findings show that quorum-sensing was induced by all the tested samples of infected sugar canes, date palm tree pieces, and date palm trees. While, cytotoxicity was induced only by infected date palm tree pieces, and genotoxicity was induced only by infected date palm trees. The bacterial monitoring results enable the identification of specific signatures that will allow a quick and accurate diagnosis.

摘要

红棕象甲(RPW),又称锈色棕榈象,被认为是棕榈科植物的严重害虫。通常情况下,感染的早期阶段没有明显的迹象。一种有吸引力的早期非可见感染感应方法是基于挥发性有机化合物(VOCs)。在这项研究中,使用全细胞细菌生物传感器来识别枣椰树(Phoenix dactylifera)中的 RPW。这些细胞经过基因改造,在存在一般应激时会产生光。生物发光细菌检测面板基于三种经过基因工程改造的大肠杆菌菌株,它们对细胞毒性(TV1061)、遗传毒性(DPD2794)或群体感应(K802NR)敏感。生物发光细菌检测面板检测 VOC 的存在,然后产生光信号的变化,反映枣椰树的健康状况。生物报告细菌细胞被固定在海藻酸钙片剂中,并放置在密封的罐子中,与测试样品没有直接接触,从而使它们仅暴露于周围空气中的 VOC。将固定化细菌细胞暴露于受 RPW 感染或未受感染的甘蔗、枣椰树切片和枣椰树附近的空气中。使用商用平板读数仪进行信号测量。研究结果表明,群体感应被所有受感染的甘蔗、枣椰树切片和枣椰树测试样本所诱导。然而,细胞毒性仅由受感染的枣椰树切片诱导,遗传毒性仅由受感染的枣椰树诱导。细菌监测结果能够识别出特定的特征,从而实现快速准确的诊断。

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