菠菜土壤中典型抗生素的微生物降解及细菌多样性

doi:10.3864/j.issn.0578-1752.2018.19.011

摘要/Abstract

摘要： 【目的】研究抗生素在土壤中残留情况及其对土壤细菌多样性的影响，为今后抗生素类污染物的土壤微生物修复和环境风险评价提供依据。【方法】通过在种植菠菜土壤中添加抗生素的盆栽试验，以庆大霉素和泰乐菌素及对应功能降解微生物（庆大霉素降解真菌FZC3和泰乐菌素降解细菌无色杆菌）为研究对象，设计7个处理，分别为庆大霉素（CG）、庆大霉素+FZC3（CGF）、泰乐菌素（CT）、泰乐菌素+无色杆菌（CTW）、两种抗生素（CM）、两种抗生素+两种降解菌（CMM）和空白（CC），其中庆大霉素残留率的对比组为CG、CGF、CM和CMM处理；泰乐菌素残留率的对比组为CT、CTW、CM和CMM处理。借助固相萃取-液质联用及Illumina高通量测序技术，对试验过程中抗生素残留及其对土壤细菌多样性影响进行研究。【结果】添加FZC3和无色杆菌可分别显著提高土壤中庆大霉素和泰乐菌素的去除效果，但随着菠菜的生长，各处理间抗生素残留率差异逐渐变小：第一周不同处理抗生素残留率差异最为显著，其中CMM处理中庆大霉素的残留率最低，为53.93%；CTW处理中泰乐菌素的残留率最低，为3.92%。第一周CGF处理比CG处理中庆大霉素残留率下降了约3.3%，CTW处理比CT处理中泰乐菌素的残留率降低了4.1%。同时，添加抗生素及其降解菌会不同程度影响土壤细菌丰富度和多样性：与CC相比，CG、CTW和CMM 3个处理中土壤细菌的丰富度和多样性有显著差异（P＜0.05）。FZC3可以缓解庆大霉素对土壤细菌的抑制作用，与CC处理相比，CG处理马赛菌和芽胞杆菌的相对丰度更高，而CGF与CC组群落结构相似；无色杆菌对土壤细菌菌群的影响程度大于其对缓解泰乐菌素毒性抑制作用的影响，相较于CT和CC处理，CTW处理中无色杆菌的添加抑制了土壤其他细菌的生长，而CT与CC处理各属细菌菌群相对丰度差异不显著。CM与CC处理相比，土壤中细菌丰富度、多样性和细菌群落结构均无显著差异，表明两种抗生素同时添加其相互间呈现拮抗作用。【结论】土壤中庆大霉素和泰乐菌素的残留对土壤微生物群落结构有不同程度的影响。添加功能微生物可以提高两种抗生素的去除效率，FZC3添加可以缓解庆大霉素对土壤细菌的影响，而无色杆菌添加对土壤细菌菌群影响较大。

关键词: 庆大霉素, 泰乐菌素, 抗生素降解菌, 细菌多样性, 菠菜

Abstract: 【Objective】 To provide an efficiently and eco-friendly solution for elimination of antibiotics in the soils and find a foundation for the evaluation of antibiotics contaminated land, this study was carried out about the residue of antibiotics in soils and their impacts on the abundance, diversity and composition of soil bacterial community. 【Method】 In this study, two types of antibiotics (gentamicin and tylosin) and two antibiotic-degrading microbe (a fungus named Aspergillus Terreus FZC3 to degrading gentamicin, a bacterium named Achromobacter to degrading tylosin) were selected to mix in spinach-growing soil. There were 7 treatments in total, including gentamicin (CG), gentamicin and FZC3 (CGF), tylosin (CT), tylosin and Achromobacter (CTW), two antibiotics (CM), two antibiotics and their degrading microbe (CMM), and blank control (CC). Solid phase extraction and HPLC-MS/MS technology were employed to study the impacts of degrading-microbe on antibiotics. The interactive influence of antibiotics and their degrading-microbe on soil bacterial community were studied by Illumina high-throughput sequencing technologies. 【Result】 Antibiotic-degrading microbe could improve the removal of the two antibiotics in the soil, but as time went by, the differences in the residual rates of antibiotics among all the treatments gradually minimized. During the entire test period, the differences of antibiotics residual rates between treatments were most significant in the first week. Under the CMM treatment, which spiked with two antibiotics and their degrading microbe, the residual rate of gentamicin was lowest (53.93%). Under the CTW treatment, which only spiked tylosin and Achromobacter, the residual rate of tylosin was as low as 3.92%. In the first week, the residual rate of gentamicin under the CGF treatment (spiked with gentamicin and FZC3) decreased about 3.3% compared with the CG treatment (spiked with gentamicin), and the residual rate of tylosin under the CTW treatment decreased about 4.1% compared with the CT treatment (spiked with tylosin). According to the results of high-throughput sequencing, there were significant differences (P＜0.05) between the bacterial richness and diversity in the three treatments of CG, CTW and CMM, compared with CC. Comparing the samples of the first week, the relative abundances of Massilia, Bacillus and etc. under the CG treatment were significantly lower than those in CC treatment; the relative abundance of Massilia in CT treatment was significantly lower than that in the CC treatment; but the decrease of inhibitory effects of antibiotics on these bacteria in the second week, indicating that gentamicin and tylosin in the early period of pot experiment had a greater impacts on soil bacterial communities. The bacterial community structure under the CGF treatment was similar to that under the CC treatment within the fortnight, indicating that gentamicin-degrading fungus FZC3 could alleviate the inhibitory effects of gentamicin on bacteria. In contrast with CT treatment, the relative abundance of Achromobacter was significantly increased while other genus decreased under the CTW treatment, indicating that the tylosin-degrading bacteria had a greater impact on the bacterial community structure. There was no significant difference in bacterial richness, diversity and community structure within fortnight between CC treatment and CM treatment (spiked with two antibiotics), indicating that there might be an antagonistic effect in soil bacteria when gentamicin and tylosin were added simultaneously. 【Conclusion】 Residues of gentamicin and tylosin in soil had different degrees of influence on bacterial abundance, diversity, and structure in soils. The addition of exogenous antibiotic-degrading microbe would not only speed up the removal efficiency of the two antibiotics, but also alleviate the effect of antibiotics on soil bacterial communities to some extent.

Key words: gentamicin, tylosin, antibiotic-degrading microbe, bacterial diversity, spinach

章程，冯瑶，刘元望，成登苗，郑宇，李兆君. 菠菜土壤中典型抗生素的微生物降解及细菌多样性[J]. 中国农业科学, 2018, 51(19): 3736-3749.

ZHANG Cheng, FENG Yao, LIU YuanWang, CHENG DengMiao, ZHENG Yu, LI ZhaoJun. The Degradation of Typical Antibiotics and Their Effects on Soil Bacterial Diversity in Spinach Soil[J]. Scientia Agricultura Sinica, 2018, 51(19): 3736-3749.

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