| 蒋宇,刘单鹏,汤宇晴,赵升,熊轩,严贵丽,欧阳凯.不同黏土矿物对恶臭假单胞菌KT2440生物膜形成的影响机制[J].农业环境科学学报,2025,44(12):3162-3171. |
| 不同黏土矿物对恶臭假单胞菌KT2440生物膜形成的影响机制 |
| Mechanism of different clay minerals influencing the biofilm formation of Pseudomonas putida KT2440 |
| 投稿时间：2025-03-05 |
| DOI：10.11654/jaes.2025-0220 |
| 中文关键词: 黏土矿物 恶臭假单胞菌 生物膜 氧化应激反应 代谢活性 |
| 英文关键词: clay minerals Pseudomonas putida biofilm oxidative stress response metabolic activity |
| 基金项目:湖南省自然科学基金面上项目（2024JJ5202）；湖南省教育厅重点项目（24A0163）；国家自然科学基金青年基金项目（42007021）；中国博士后科学基金项目（2020M672498） |
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| 中文摘要: |
| 为揭示土壤中黏土矿物对细菌生物膜生长发育的影响机制，本研究借助多功能酶标仪、荧光显微镜等科学仪器，结合结晶紫染色法、荧光检测等技术手段，探讨了3种典型黏土矿物（蒙脱石、高岭石、针铁矿）对恶臭假单胞菌KT2440细胞生长及生物膜形成的影响。结果表明：相较于纯培养体系，蒙脱石处理体系中细菌生物膜生物量增加了78.95%，低浓度阈值内的针铁矿（0~5mg·L-1）和高岭石（0~100 mg·L-1）对细菌生物膜的形成均有不同程度的促进作用，而高浓度针铁矿（>5 mg·L-1）和高岭石（>100 mg·L-1）处理体系则显著抑制细菌生物膜形成，最高抑制率分别达83.78%和34.72%。通过共沉降试验和荧光显微镜观察发现，针铁矿能有效吸附并杀死细菌。针铁矿、高岭石诱导产生的活性氧（ROS）含量分别是纯培养体系的3.66倍和1.97倍；针铁矿在与恶臭假单胞菌共培养 192 h后，细胞内三磷酸腺苷（ATP）含量相比纯培养体系降低了 58.21%。蒙脱石通过静电互斥作用减少对细菌细胞膜的损伤，并利用高比表面积促进细胞定殖，显著增强其生长与生物膜形成。针铁矿和高岭石在低浓度时可以杀死部分细胞，释放营养物质促进生物膜形成，而高浓度则通过诱发氧化应激破坏细胞代谢功能，抑制生物膜发育。 |
| 英文摘要: |
| To elucidate the influence mechanisms of clay minerals in soil on the growth and development of bacterial biofilms. This study employed scientific instruments including a multifunctional microplate reader and fluorescence microscope, combined with technical approaches such as crystal violet staining and fluorescence detection, to investigate the effects of three typical soil clay minerals （montmorillonite, kaolinite, and goethite）on the growth of Pseudomonas putida KT2440 and biofilm formation. The results demonstrated that, compared to the pure culture system, the biofilm biomass in the montmorillonite-treated system increased by 78.95%. Within low concentration thresholds, goethite（0-5 mg·L-1）and kaolinite（0-100 mg·L-1）both promoted bacterial biofilm formation to varying degrees, while high concentrations of goethite（>5 mg·L-1）and kaolinite（>100 mg·L-1）significantly inhibited biofilm formation, with maximum inhibition rates of 83.78% and 34.72%, respectively. Through co-sedimentation experiments and fluorescence microscopy observations, it was demonstrated that goethite can effectively adsorb and kill bacteria. The production of reactive oxygen species（ROS） induced by goethite and kaolinite was 3.66 times and 1.97 times higher, respectively, than that in the pure culture system. Additionally, after 192 h of co-cultivation with P. putida, the intracellular adenosine triphosphate（ATP） content in the goethite-treated system decreased by 58.21% compared to the pure culture system. Montmorillonite mitigates bacterial cell membrane damage through electrostatic repulsion and enhances bacterial colonization via its high specific surface area, thereby significantly promoting bacterial growth and biofilm formation. In contrast, goethite and kaolinite at low concentrations kill a subset of cells, releasing nutrients that stimulate biofilm formation, whereas at high concentrations, they induce oxidative stress, disrupt cellular metabolic functions, and suppress biofilm development. |
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