Effects and mechanisms of herbicides on horizontal transfer of antibiotic resistance genes through plasmid-mediated conjugation
SUN Feng-fei1, WANG Xiu2, MAO Yu-meng1, LIU Ke2, LIU Juan1
1. Institute of Organic Contaminant Control and Soil Remediation, College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, China; 2. Institute of Animal Husbandry and Poultry Science, Nanjing 210017, China
摘要 The spread of antibiotic resistance genes (ARGs) in farmland ecosystem is directly related to human health, and has attracted much attention in recent years. Herbicides are common chemical reagents widely used in farmland ecosystem, and until now, little information has been known about its effects and mechanisms on horizontal gene transfer (HGT) of ARGs between bacteria. In this study, with Escherichia coli DH5α(carrying RP4 plasmid with TetR, AmpR, KanR) as the donor and E. coli HB101as the recipient, a series of diparental conjugation experiments were conducted to investigate the effects of butachlor, paracetamol and glufosinate on HGT of ARGs via plasmid-mediated conjugation. Furthermore, the mechanisms involved were also clarified from cell membrane permeability, intracellular ROS content, and the transcription levels of genes related to conjugation. The results showed that 100~800mg/L of butachlor and 40~160mg/L of paracetamol could promote the conjugation transfer of plasmid RP4. Under test concentrations, butachlor stimulated the intracellular production of ROS, while paraquat reduced the ROS level. Under the treatment of butachlor or paracetamol, the surface of the recipient bacterium appeared obvious shrinkage, and with the increase of concentration, the pore and even break appeared. Butachlor (400~800mg/L) and paraquat (40~80mg/L) could both increase cell membrane permeability, inhibit the korA transcription and promote the transcription of trbBp、trfAp and traA, thus promoting the conjugation transfer of RP4. Under test concentrations, there was only a little effect of glufosinate on RP4 conjugation, as well as cell membrane permeability, intracellular ROS content and transcription of conjugation-related regulatory genes. This study indicated that widespread use of herbicides in agricultural production could affect the HGT of ARGs via plasmid-mediated conjugation, and the application of reasonable types and concentrations of herbicides could reduce the ARG transmission in both soil environments and plants.
Abstract：The spread of antibiotic resistance genes (ARGs) in farmland ecosystem is directly related to human health, and has attracted much attention in recent years. Herbicides are common chemical reagents widely used in farmland ecosystem, and until now, little information has been known about its effects and mechanisms on horizontal gene transfer (HGT) of ARGs between bacteria. In this study, with Escherichia coli DH5α(carrying RP4 plasmid with TetR, AmpR, KanR) as the donor and E. coli HB101as the recipient, a series of diparental conjugation experiments were conducted to investigate the effects of butachlor, paracetamol and glufosinate on HGT of ARGs via plasmid-mediated conjugation. Furthermore, the mechanisms involved were also clarified from cell membrane permeability, intracellular ROS content, and the transcription levels of genes related to conjugation. The results showed that 100~800mg/L of butachlor and 40~160mg/L of paracetamol could promote the conjugation transfer of plasmid RP4. Under test concentrations, butachlor stimulated the intracellular production of ROS, while paraquat reduced the ROS level. Under the treatment of butachlor or paracetamol, the surface of the recipient bacterium appeared obvious shrinkage, and with the increase of concentration, the pore and even break appeared. Butachlor (400~800mg/L) and paraquat (40~80mg/L) could both increase cell membrane permeability, inhibit the korA transcription and promote the transcription of trbBp、trfAp and traA, thus promoting the conjugation transfer of RP4. Under test concentrations, there was only a little effect of glufosinate on RP4 conjugation, as well as cell membrane permeability, intracellular ROS content and transcription of conjugation-related regulatory genes. This study indicated that widespread use of herbicides in agricultural production could affect the HGT of ARGs via plasmid-mediated conjugation, and the application of reasonable types and concentrations of herbicides could reduce the ARG transmission in both soil environments and plants.
孙凤飞, 王秀, 毛雨梦, 刘珂, 刘娟. 除草剂对质粒介导ARGs接合转移的影响及机制[J]. 中国环境科学, 2024, 44(11): 6426-6434.
SUN Feng-fei, WANG Xiu, MAO Yu-meng, LIU Ke, LIU Juan. Effects and mechanisms of herbicides on horizontal transfer of antibiotic resistance genes through plasmid-mediated conjugation. CHINA ENVIRONMENTAL SCIENCECE, 2024, 44(11): 6426-6434.
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