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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2014, Vol. 40 Issue (1): 75-84    DOI: 10.3785/j.issn.1008-9209.2013.06.141
资源与环境科学     
外源抗生素对茶园土壤微生物群落结构的影响
徐晨光， 张奇春*， 侯昌萍
（浙江大学环境与资源学院，浙江省亚热带土壤与植物营养重点研究实验室，杭州 310058）
 Effect of exogenous antibiotics on soil microbial community structure in tea garden
Xu Chenguang,  Zhang Qichun*，  Hou Changping
(Zhejiang Provincial Key Laboratory of Subtropical Soil and Plant Nutrition, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310058,China)
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摘要： 采用室内恒温培养，通过向茶园土壤中分别添加2种典型抗生素（青霉素和四环素）溶液的方法研究了不同质量分数的外源抗生素对茶园土壤微生物生态特性的影响，以评价其环境生态效应。结果表明：外源青霉素的添加对茶园土壤细菌、放线菌和真菌均有一定的抑制作用，随着青霉素质量分数的增加抑制作用也增加。当青霉素质量分数为100 mg/kg时，培养3周后与对照（CK）相比，细菌、真菌和放线菌的数量分别减少了80%、50%和50%。四环素在低质量分数下即可显著抑制茶园土壤中细菌和放线菌的生长，但对真菌没有明显的抑制作用。培养4周时，青霉素和四环素的抑制作用减弱，土壤微生物表现出对抗生素的抗性。磷脂脂肪酸(phospholipid fatty acid, PLFA)法的结果表明，用不同质量分数的青霉素或四环素处理的茶园土壤中微生物群落结构发生明显变化。青霉素或四环素处理后G-/G+比值都升高，表明青霉素和四环素对于茶园土壤中革兰阳性菌的抑制作用强于革兰阴性菌。四环素处理后磷脂脂肪酸20:0的含量最丰富，表明磷脂脂肪酸20:0表征的微生物可能是茶园土壤产生四环素耐性菌的主要类群。
Abstract: Antibiotics are medical drugs used to kill bacteria and treat infections. But in recent years, more and more antibiotics have been used in animal feed as growth promoters. Manure is used worldwide in agricultural practice as a fertilizer to enhance plant yields. Several studies revealed that manure also had a stimulating effect on the soil microbial biomass and community structure. However, in modern agriculture, the application of manure to field is very often combined with the spread of veterinary antibiotics. Tetracycline and penicillin are possible soil contaminants, since they are able to reach agricultural soils with manure of treated farm animals.
Zhejiang Province is a major tea producing region in China, whose tea exports amount to 100 000 tons and  production value accounts for one-third of the country. Every year a large number of organic fertilizers are put into the tea garden and the spread of veterinary antibiotics increases with applying organic fertilizer. The organic fertilizer can increase the yield of tea tree, but  the harm to soils caused by antibiotics  can not be avoided  in the process of using organic fertilizer. Microorganism is a promoter of soil nutrient transformation, which  plays an important role in maintaining balance, health and high quality of soil ecosystem. Although it has been confirmed that organic fertilizer (especially manure) application could promote the growth of crops, the impact of antibiotics by applying organic fertilizer on soil microbial community is still unknown. Thus, the aim of this research is to investigate possible toxic effects of antibiotic exposure on the indigenous bacterial community of tea garden.
Topsoils (020 cm) were collected from Meijiawu Tea Village, Hangzhou, Zhejiang Province. Soils were spiked with a range of tetracycline or penicillin concentrations. The basic physicochemical properties of the tested soil were measured by routine analysis methods. The amounts of bacteria, fungi and actinomycetes of different treatments were calculated by plate count method. As we know, only 0.1%10% microbes can be cultivated by this method, therefore we also used the method of phospholipid fatty acid (PLFA) to analyze the PLFAs contents to make a quantitative description of microbial community. After cultivation for three weeks, compared with the control treatment (CK), the amounts of bacteria, fungi and actinomycetes in P5 treatment (100 mg/kg) decreased by 80%, 50% and 50%, respectively. Soil microorganisms showed resistance to antibiotics after incubation for four weeks. The results of phospholipid fatty acid method  showed that soil microbial community structure changed obviously under different contents of antibiotics treatments. The ratio of G-/G+ increased after antibiotic treatments. Principal component analysis showed different treatments located in four different quadrants. The content of PLFA 20:0 was rich in T3, T4 and T5 treatments.
It is concluded that exogenous penicillin has certain inhibitory effect to bacteria, fungi and actinomycetes, and the inhibition increases with penicillin concentration. Tetracycline inhibits the growth of bacteria and actinomycetes in tea garden soil, but shows no obvious inhibitory effect against fungi. Inhibition of Gram-negative bacteria for both penicillin and tetracycline are stronger than Gram-positive bacteria. There is great difference in soil microbial community structure among different treatments. Microbes characterized by PLFA 20:0 may be the main groups of anti-tetracycline microbes.
出版日期: 2014-01-20
CLC:  Q 89  
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侯昌萍
徐晨光
张奇春*

引用本文:

徐晨光， 张奇春*， 侯昌萍. 外源抗生素对茶园土壤微生物群落结构的影响[J]. 浙江大学学报(农业与生命科学版), 2014, 40(1): 75-84.

Xu Chenguang, Zhang Qichun*， Hou Changping .  Effect of exogenous antibiotics on soil microbial community structure in tea garden. Journal of Zhejiang University (Agriculture and Life Sciences), 2014, 40(1): 75-84.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2013.06.141    或    http://www.zjujournals.com/agr/CN/Y2014/V40/I1/75

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