真菌电化学修复除草剂污染土壤：降解动力学探索

doi:10.13560/j.cnki.biotech.bull.1985.2023-0895

摘要/Abstract

摘要：

【目的】 微生物燃料电池（microbial fuel cell, MFC）在去除污染物的同时产出电能，是一种颇有前景的生态修复手段。构建真菌强化MFC装置，比较电动力（EK）、真菌、MFC修复除草剂污染土壤效果及优缺点，探索MFC在有机污染物修复中的应用潜力。【方法】 设计了一种添加真菌进行生物强化的MFC，并用EK、真菌、MFC三种方法修复两种除草剂污染的灭菌土壤。经筛选和驯化的疣孢漆斑菌和踝节菌菌株用于后两种方法，研究真菌强化对MFC去除除草剂的影响。测量土壤pH、电导率、除草剂去除率，MFC产电性能，用气相色谱-质谱鉴定两种除草剂的降解产物。【结果】 EK修复中，添加模拟电解液、碳纤维条、加电10 V的处理组7 d后氯氟吡啶酯（F）和高效氟吡甲禾灵（H）去除率分别为71%和38%。真菌、MFC处理F的最大去除率达到100%。对比踝节菌，疣孢漆斑菌对两种除草剂的降解性能更好，疣孢漆斑菌、踝节菌单菌构建的MFC对H的去除率分别为62.5%和24.1%。F降解产物为氟氯吡啶酸，H降解产物为乙酸大茴香酯，推测了降解路径和降解动力学。三种方法降解F以及EK降解H均符合动力学一级反应，而真菌和MFC降解H符合二级反应。【结论】 对比EK、真菌修复，MFC修复效果更好，该方法可以较快地修复土壤又无需额外供电，是一种经济有效的自持式修复策略。

关键词: 电化学修复, 微生物燃料电池, 踝节菌DJTU-SJ5, 疣孢漆斑菌DJTU-sh7, 土壤修复

Abstract:

【Objective】 Microbial fuel cell（MFC）, which produces electricity while removing pollutants, is a promising means of ecological restoration. We aim to construct a fungal-augmented MFC device, compare the effects, advantages and disadvantages of electrokinetic（EK）, fungal and MFC remediation against herbicides in the polluted soil, and explored the application potential of MFC in the remediation of organic pollutants. 【Method】 A novel MFC with fungal bioaugmentation was designed. The contaminated soil by two herbicides was remediated by three methods of EK, fungi and MFC remediations, and the screened and domesticated Talaromyces dalianensis and Myrothecium verrucaria strains were implemented in the latter two approaches. The effect of fungal augmentation on the removal of herbicides was studied. Soil pH, electrical conductivity, herbicide removal, and MFC electrical performance were quantified, and degradation products of two herbicides were identified by gas chromatography-mass spectrometry. 【Result】 In EK remediation, when simulated electrolyte, carbon fiber and 10 V were applied for 7 d, the removal rate of florpyrauxifen-benzyl（F）and haloxyfop-P（H）was 71% and 38%, respectively. Compared with H, F was more easily degraded, and fungal and MFC treatments completely removed it. Compared with T. dalianensis, M. verrucaria strain had better degradation capacity to both herbicides. MFC based on two fungi had the removal rate of 62.5% and 24.1% respectively toward H. The degradation product of F and H was halauxifen and 4-methoxybenzyl acetate, respectively, and the degradation pathways and kinetics were speculated. The degradation of F by three methods and the degradation of H by EK followed the first-order kinetic reaction, while the degradation of H by fungi and MFC followed the second-order reaction. 【Conclusion】 Compared with EK and fungal remediations, MFC remediation had a better effect, which can remediate the soil without the need for additional power supply, making it a cost-effective self-sustaining remediation strategy.

Key words: electrochemical remediation, microbial fuel cell, Talaromyces dalianensis DJTU-SJ5, Myrothecium verrucaria DJTU-sh7, soil remediation

郝大程, 郑宇薇, 王凡, 韩蕾, 张赜. 真菌电化学修复除草剂污染土壤：降解动力学探索[J]. 生物技术通报, 2024, 40(3): 261-272.

HAO Da-cheng, ZHENG Yu-wei, WANG Fan, HAN Lei, ZHANG Ze. Fungal Electrochemical Remediation of Herbicide-contaminated Soil: Preliminary Study on Degradation Kinetics[J]. Biotechnology Bulletin, 2024, 40(3): 261-272.

图/表 9

图1 除草剂化学结构 A：氯氟吡啶酯化学结构式，C20H14Cl2F2N2O3；B：高效氟吡甲禾灵结构式，C15H11ClF3NO4

Fig. 1 Chemical structures of herbicides A: Florpyrauxifen-benzyl, C20H14Cl2F2N2O3; B: haloxyfop-P, C15H11ClF3NO4

表1 真菌与MFC降解方案设计

Table 1 Degradation scheme of fungi and MFC

分组Group	土壤中污染物含量 Pollutant content in soil	菌液Composition
（1）F	97.14 mg/kg F	不添加菌液No fungal addition
（2）F+T	97.14 mg/kg F	20 mL F+ 20 mL T
（3）F+M	97.14 mg/kg F	20 mL F+20 mL M
（4）F+T+M	97.14 mg/kg F	20 mL F+ 10 mL T + 10 mL M
（5）H	245.71 mg/kg H	不添加菌液No fungal addition
（6）H+T	245.71 mg/kg H	20 mL H+ 20 mL T
（7）H+M	245.71 mg/kg H	20 mL H+ 20 mL M
（8）H+T+M	245.71 mg/kg H	20 mL H+ 10 mL T + 10 mL M

图2 EK降解实验 A: EK修复装置。B: EK修复装置绘制，对电极与参比电极相接后1与钢网阴极3相连，工作电极2与石墨毡阳极4相连。C:土样取点剖面图，1为距离阳极 2 cm点；2为距离阳极4 cm点；3为距离阳极6 cm点（近阴极）

Fig. 2 Methods of EK degradation A: EK remediation device. B: After the counter electrode is connected with the reference electrode, 1 is connected with the steel mesh cathode 3, and the working electrode 2 is connected with the carbon felt anode 4. C: Profile of the sampling sites; 1 is 2 cm from the anode, 2 is 4 cm from the anode, and 3 is 6 cm from the anode（near the cathode）

图3 MFC降解实验 A: MFC实验装置; B: MFC实验装置绘制，阳极与阴极均水平放置，阳极位于阴极正上方，间隔3.5 cm，土壤顶部距容器上方边缘2 cm; C: MFC装置底部，设有小孔方便阴极与空气接触; D: MFC装置剖面图（1：阳极；2：阴极；A：阳极取样点；C：阴极取样点）

Fig. 3 Methods of MFC degradation A: MFC device. B: The anode and the cathode are placed horizontally. The anode is located directly above the cathode, with an interval of 3.5 cm, and the top of the soil is 2 cm from the upper edge of the container. C: At the bottom of the MFC device, a small hole is provided to facilitate the contact between the cathode and the air. D: Profile of MFC device（1: anode; 2: cathode; A: the anode sampling point; C: the cathode sampling point）

图4 EK修复结果氯氟吡啶酯（F）组：F1、F2（碳纤维2 g）为对照组不加电，F3、F4、F5、F6（碳纤维2 g）施加电压分别为5 V、10 V、10 V、10 V，其中F5组加去离子水，其他条件不变。高效氟吡甲禾灵（H）分组同F组。A: F组土壤pH变化，0点为加电前土壤pH，2 cm、4 cm、6 cm点均为加电处理后土壤pH。B: H组土壤pH 变化。C: F组土壤电导率变化，0点为加电前土壤电导率，2 cm、4 cm、6 cm点均为加电处理后土壤电导率。D: H组土壤电导率变化。E: F组电流密度变化，右上小图为电荷变化量。F: H组电流密度变化。G: F去除率。H: H去除率。不同小写字母代表同层不同处理组之间的显著性差异，P < 0.05，下同

Fig. 4 Results of EK remediation F groups: F1 and F2（+ carbon fiber 2 g）are control groups without electricity; F3, F4, F5, F6（+ carbon fiber 2 g）are subject to voltage 5 V, 10 V, 10 V, 10 V, respectively; deionized water rather than electrolyte is applied in F5 group. The grouping of H is similar to that of F. A: Soil pH changes of F groups. 0, no electrode. B: Soil pH changes of H groups. C: Changes of soil electrical conductivity in F groups. 0, no electrode. D: Changes of soil electrical conductivity in H groups. E: The change of current density in F groups, and the upper right inset shows the changes of charge. F: The change of current density in H groups. G: Removal rate of F. H: Removal rate of H. Different lowercase letters represent the significant differences（P < 0.05）between different treatment groups of the same layer, the same below

图5 真菌修复 A: F组土壤pH变化。B: H组土壤pH变化。C: F组土壤电导率变化。D: H组土壤电导率变化。E: F去除率。F: H去除率。不同小写字母代表同一时间不同处理之间的显著性差异，P < 0.05，下同

Fig. 5 Fungal remediation A: Soil pH changes of F groups. B: Soil pH changes of H groups. C: Changes of soil electrical conductivity in F groups. D: Changes of soil electrical conductivity in H groups. E: Removal rate of F. F: Removal rate of H. Different lowercase letters indicate the significant differences（P < 0.05）between different treatment groups of the same duration

图6 MFC修复 A为阳极，C为阴极。F组MFC电池性能：A：第2天。B:第15天。C:第30天。H组MFC电池性能：D:第2天。E:第15天。F:第30天。G: F组土壤pH变化。H: H组土壤pH变化。I: F组土壤电导率变化。J: H组土壤电导率变化。K: F去除率。L: H去除率

Fig. 6 Results of MFC remediation A: Anode; C: cathode. MFC performance of F groups: A: Day 2. B: Day 15. C: Day 30. MFC performance of H groups: D: Day 2. E: Day 15. F: Day 30. G: Changes of soil pH in F groups. H: Changes of soil pH in H groups. I: Changes of soil electrical conductivity in F groups. J: Changes of soil electrical conductivity in H groups. K: Removal rate of F. L: Removal rate of H

图7 除草剂降解产物 A: F降解产物质谱图; B: H降解产物质谱图; C:氟氯吡啶酸全扫描离子碎片图; D:乙酸大茴香酯全扫描离子碎片图; E: 氟吡甲禾灵全扫描离子碎片图; F: F降解路径; G: H降解路径

Fig. 7 Degradation products of herbicides A: Mass spectrum of F degradation products. B: Mass spectrum of H degradation products. C: Full scan ion fragment diagram of halauxifen. D: Full scan ion fragment diagram of 4-methoxybenzyl acetate. E: Full scan ion fragment diagram of haloxyfop-methyl. F: Degradation pathway of F. G: Degradation pathway of H

表2 除草剂降解动力学拟合

Table 2 Degradation kinetics of herbicides

处理 Treatment	氯氟吡啶酯 Florpyrauxifen-benzyl（F）				高效氟吡甲禾灵Haloxyfop-P（H）
处理 Treatment	反应 Reaction	降解速率常数 Degradation rate constant	R²	半衰期Half-life/d	反应 Reaction	降解速率常数 Degradation rate constant	R²	半衰期 Half-life/d
EK处理 EK treatment	一级反应First order reaction	0.248 ± 0.009	0.993 01	2.8	一级反应First order reaction	0.053 ± 0.002	0.991 44	13.0
微生物处理 Microbial treatment	一级反应First order reaction	0.054 ± 0.001	0.946 30	12.7	二级反应Second order reaction	1.968×10-4 ± 4.979×10-7	0.999 99	50.6
MFC处理 MFC treatment	一级反应First order reaction	0.136 ± 0.001	0.998 16	5.1	二级反应Second order reaction	5.921×10-4 ± 9.888×10-7	0.999 99	16.8

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