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首页> 外文期刊>Environmental Science & Technology >Analysis of RNA Interference (RNAi) Biopesticides: Double-Stranded RNA (dsRNA) Extraction from Agricultural Soils and Quantification by RT-qPCR
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Analysis of RNA Interference (RNAi) Biopesticides: Double-Stranded RNA (dsRNA) Extraction from Agricultural Soils and Quantification by RT-qPCR

机译：RNA干扰（RNAi）生物农药的分析：从农业土壤中提取双链RNA（dsRNA）并通过RT-qPCR进行定量

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摘要

Double-stranded RNA (dsRNA) molecules are used as a novel class of biopesticides. To enable assessments of the ecological risk associated with their release to receiving environments, we developed an approach to quantify dsRNA in agricultural soils using quantitative reverse transcription-polymerase chain reaction (RT-qPCR). To allow quantification of dsRNA adsorbed to particles, we also developed a protocol to transfer dsRNA from particles to the extraction buffer by changing particle surface charge and adding constituents to compete with dsRNA for adsorption sites. Our approach could quantify dsRNA amounts as low as 0.003 ng_(dsRNA)/g_(soil). This approach is the first available field-applicable approach able to quantify dsRNA biopesticides down to environmentally relevant concentrations. We applied this approach to investigate dsRNA dissipation (including dilution, degradation, and adsorption) in two agricultural soils. When we applied a low amount of dsRNA (1 ng_(dsRNA)/g_(soil) to the soils, we observed that a greater fraction of dsRNA was adsorbed to and extractable from soil particles in a silty clay loam soil than in a fine sandy loam soil. In both soils, dsRNA dissipated on the timescale of hours. Overall, these results demonstrate that our approach can be applied to assess the environmental fate of dsRNA biopesticides at concentrations relevant to their release to soils.
机译：双链RNA（dsRNA）分子被用作一类新型的生物农药。为了评估与其释放到接收环境相关的生态风险，我们开发了一种使用定量逆转录聚合酶链反应（RT-qPCR）定量农业土壤中dsRNA的方法。为了量化吸附到颗粒上的dsRNA，我们还开发了一种协议，可通过改变颗粒表面电荷并添加与dsRNA竞争吸附位点的成分，将dsRNA从颗粒转移到提取缓冲液中。我们的方法可以量化低至0.003 ng_（dsRNA）/ g_（土壤）的dsRNA量。这种方法是第一个能够将dsRNA生物农药定量降低到与环境有关的浓度的可现场应用的方法。我们应用这种方法研究了两种农业土壤中dsRNA的耗散（包括稀释，降解和吸附）。当我们向土壤中施用少量dsRNA（1 ng_（dsRNA）/ g_（土壤））时，我们观察到，与细沙质土壤相比，粉质粘土壤土中的颗粒吸附和提取的dsRNA比例更高。壤土：在这两种土壤中，dsRNA均在数小时的时间内消散，总体而言，这些结果表明，我们的方法可用于评估dsRNA生物农药在与土壤中释放相关的浓度下的环境状况。

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  • 来源
    《Environmental Science & Technology》 |2020年第8期|4893-4902|共10页
  • 作者

    Ke Zhang; Jingmiao Wei; Kara E. Huff Hartz; Michael J. Lydy; Tae Seok Moon; Michael Sander; Kimberly M. Parker;

  • 作者单位

    Department of Energy Environmental & Chemical Engineering Washington University in St. Louis St. Louis Missouri 63130 United States;

    Center for Fisheries Aquaculture and Aquatic Sciences Department of Zoology Southern Illinois University Carbondale Illinois 62901 United States;

    Department of Energy Environmental & Chemical Engineering Washington University in St. Louis St. Louis Missouri 63130 United States;

    Department of Environmental Systems Science (DUSYS) ETH Zurich 8092 Zurich Switzerland;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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  • 正文语种 eng
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