A MnO2 nanosheet-mediated CRISPR/Cas12a system for the detection of organophosphorus pesticides in environmental water.

Nowadays, easy, convenient, and sensitive sensing strategies are still critical for organophosphorus pesticides in environmental water samples. Herein, a novel organophosphorus pesticide (OP) assay based on acetylcholinesterase (AChE) and a MnO2 nanosheet-mediated CRISPR/Cas12a reaction is reported. The single-strand DNA (ssDNA) activator of CRISPR/Cas12a was simply adsorbed on the MnO2 nanosheets as the nanoswitches of the assay. In the absence of target OPs, AChE hydrolyzed acetylcholine (ATCh) to thiocholine (TCh), which reduced the MnO2 nanosheets to Mn2+, resulting in the release of the activator followed by activation of the CRISPR/Cas12a system. The activated Cas12a thereafter nonspecifically cleaved the FAM/BHQ1-labeled ssDNA (FQ-reporter), producing a fluorescence signal. Upon the addition of target OPs, the hydrolysis of ATCh by AChE was inhibited owing to OPs combining with AChE, and thus effective quantification of OPs could be achieved by measuring the fluorescence changes of the system. As a proof of concept, dichlorvos (DDVP) was chosen as a model OP analyte to address the feasibility of the proposed method. Attributed to the excellent trans-cleavage activity of Cas12a, the fluorescent biosensor exhibits a satisfactory limit of detection (LOD) for DDVP at 0.135 ng mL-1. In addition, the excellent recoveries for the detection of DDVP in environmental water samples demonstrate the applicability of the proposed assay in real sample research.

[Effects of soil moisture on microbial processes of soil nitrogen gases production under anaerobic conditions]. / 土壤水分对土壤产生气态氮的厌氧微生物过程的影响.

Gaseous nitrogen (N) emission [nitric oxide (NO), nitrous oxide (N2O), and nitrogen (N2)] is an important pathway of soil N loss. Nitrification and denitrification are the main processes of gaseous N production in soil. However, the contribution of heterotrophic nitrification, co-denitrification, and anammox to gaseous N production remains uncertain. In a laboratory soil incubation experiment, we used the 15N labelling and pairing technique, combining the nitrification inhibitor dicyandiamide (DCD), to quantify the contribution of different microbial processes to soil NO, N2O and N2 production under anaerobic conditions. The results showed that after 24 h anaerobic incubation, the highest total 15N recovery of three gases occurred at 65% water filled pore space (WFPS), accounting for 20.0% of total added 15N. Denitrification contributed 49.9%-94.1%, 29.0%-84.7%, and 58.2%-85.8% to the production of NO, N2O and N2 respectively, suggesting that denitrification was the predominant process of those three N gases emission. Heterotrophic nitrification was an important pathway of NO and N2O production, particularly at conditions with low soil water content (10% WFPS), with its contribution to those two N gases production being 50.1% and 42.8%, respectively. Co-denitrification contributed 10.6%-30.7% of N2O production. For N2 production, the total contribution of co-denitrification and anammox was 14.2%-41.8%. The role of co-denitrification can not be ignored for N2O and N2 production. Our results demonstrated that the 15N labelling and pairing technique is a promising tool to quantify the contribution of different microbial processes to gaseous N loss.