邹勇,
孙永学,
国家兽药安全评价(环境评估)实验室(华南农大), 国家兽医微生物耐药性风险评估实验室, 广东省兽药研制与安全评价重点实验室, 广州 510642
作者简介: 林曼霞(1992-),女,硕士研究生,研究方向为兽医药理与毒理学研究,E-mail:1058030702@qq.com.
通讯作者: 孙永学,sunyx@scau.edu.cn
基金项目: 国家自然科学基金(31772803)；广东省自然科学基金(2016A030311029)中图分类号: X171.5
Effects of Oxytetracycline on the Transcriptional Characteristics of Dominant Denitrification Genes and Flora Structure Characteristics in Indoor Manure -Soil Model
Lin Manxia,Zou Yong,
Sun Yongxue,
National Laboratory of Safety Evaluation(Environmental Assessment) of Veterinary Drugs(SCAU), National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Origin Bacteria, Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Assessment, Guangzhou 510642, China
Corresponding author: Sun Yongxue,sunyx@scau.edu.cn
CLC number: X171.5
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摘要:反硝化作用在土壤氮素循环中扮演着重要角色,而国内研究抗生素残留对反硝化作用的报道仍然较少。本实验通过构建室内抗生素-粪便-土壤模型,以研究空白组(0 μg·kg-1)、低浓度组(20 μg·kg-1)、中浓度组(200 μg·kg-1)和高浓度组(2 000 μg·kg-1)土霉素对施加鸡粪后土壤中反硝化作用的影响。利用荧光定量PCR(qPCR)和限制性末端片段长度多态性(T-RFLP)方法分析反硝化基因(nirS、nirK、narG和nosZ)的响应特征及反硝化菌群结构特点。qPCR定量结果表明低、中浓度土霉素对携带nirS、nirK基因的菌群有促进作用,而高浓度组则表现为抑制作用,而携带nosZ基因的菌群对土霉素的胁迫反应较迟钝。T-RFLP结果显示不同浓度土霉素并没有改变携带nirS基因和nosZ基因的优势菌群结构,而随着暴露时间的延长,携带nirS基因的反硝化优势片段在第30~60天之间发生了变化。这些结果表明不同浓度土霉素可以影响反硝化菌群丰度,但是却无法影响优势菌群,而土壤中其他因素可能对优势菌群的影响更大。
关键词: 土霉素/
反硝化作用/
荧光定量/
T-RFLP
Abstract:Denitrification plays an important role in soil nitrogen cycling. However, there are still few reports on effects of antibiotic residues on denitrification in China. The present study constructed an indoor manure-soil model by adding different concentrations of oxytetracycline. Four groups in different concentrations of oxytetracycline were set up:blank group (0 μg·kg-1), low concentration group (20 μg·kg-1), medium concentration group (200 μg·kg-1), high concentration group (2 000 μg·kg-1) so as to explore the effect of oxytetracycline on denitrification in soil after chicken manure application. The response of denitrifying genes (nirS, nirK, narG and nosZ) and the denitrifying microbial community were analyzed by Real-time Quantitative (qPCR) and Terminal Restriction Fragment Length Polymorphism (T-RFLP). qPCR results showed that low and medium concentrations of oxytetracycline promoted the abundance of bacteria carrying nirS and nirK genes, whereas the high concentration posed inhibitory effects on these bacteria. The bacteria carrying nosZ gene was less responsive to oxytetracycline stress. T-RFLP results showed that different concentrations of oxytetracycline did not change the dominant bacteria carrying nirS gene and nosZ gene. The dominant denitrification fragment carrying nirS gene had changed between 30 and 60 days in the extension of exposure time. These results indicated that different concentrations of oxytetracycline could affect the abundance of denitrifying bacteria, but could not affect the abundance of dominant bacteria.
Key words:oxytetracycline/
denitrification/
fluorescence quantitative/
T-RFLP.
