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一种基于简单氧化石墨烯的植物病原菌 DNA 纯化策略。

A simple graphene oxide-based DNA purification strategy for plant pathogen detection.

机构信息

Key Laboratory of Agricultural Microbiology, College of Agriculture, Guizhou University, Guiyang, China.

Institute of Vegetable Industry Technology Research, Guizhou University, Guiyang, China.

出版信息

Pest Manag Sci. 2024 Jul;80(7):3516-3525. doi: 10.1002/ps.8056. Epub 2024 Mar 15.

Abstract

BACKGROUND

The on-site molecular detection of plant pathogens is particularly important for the development of sustainable agriculture. Extracting DNA from plant tissues, microbes or coexisting environments is complex, labor-intensive and time-consuming. To facilitate this process, we propose a DNA purification strategy based on graphene oxide (GO).

RESULTS

The excellent adsorption ability of GO was verified by visualizing changes in its microscopic surface and macroscopic mixture. To further optimize the DNA purification, we determined the optimal GO concentration and treatment time at 95 °C (2 mg mL and 2 min, respectively). We confirmed that our strategy is effective on plant tissues and various microorganisms, and that the obtained DNA can be directly used for polymerase chain reaction amplification. Combining the proposed GO-based DNA purification method with the loop-mediated isothermal amplification method is superior, in terms of the required steps, time, cost and detection effect, to the cetyltrimethylammonium bromide method and a commercial kit for detecting plant pathogens.

CONCLUSION

We present a feasible, rapid, simple and low-cost DNA purification method with high practical value for scientific applications in plant pathogen detection. This strategy can also provide important technical support for future research on plant-microbial microenvironments. © 2024 Society of Chemical Industry.

摘要

背景

现场检测植物病原体对于可持续农业的发展尤为重要。从植物组织、微生物或共存环境中提取 DNA 既复杂又费力且耗时。为了简化这一过程,我们提出了一种基于氧化石墨烯(GO)的 DNA 纯化策略。

结果

GO 的优异吸附能力通过可视化其微观表面和宏观混合物的变化得到验证。为了进一步优化 DNA 纯化,我们确定了在 95°C 下的最佳 GO 浓度和处理时间(分别为 2mg/mL 和 2min)。我们证实,我们的策略对植物组织和各种微生物都有效,并且获得的 DNA 可直接用于聚合酶链反应扩增。与溴化十六烷基三甲铵法和用于检测植物病原体的商业试剂盒相比,将基于 GO 的 DNA 纯化方法与环介导等温扩增方法相结合,在所需步骤、时间、成本和检测效果方面具有优势。

结论

我们提出了一种可行、快速、简单且低成本的 DNA 纯化方法,对于植物病原体检测的科学应用具有很高的实用价值。该策略还可为未来的植物-微生物微环境研究提供重要的技术支持。 © 2024 化学工业协会。

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