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| 荧光原位杂交技术检测土壤中博德特氏菌探针的设计与应用 | |
| 引用本文： | 王芳,M.Schmi,蒋新,R.Schroll.荧光原位杂交技术检测土壤中博德特氏菌探针的设计与应用[J].土壤学报,2010,47(1):51-57. |
| 作者姓名： | 王芳 M.Schmi 蒋新 R.Schroll |
| 作者单位： | 1. 土壤与农业可持续发展国家重点实验室,(中国科学院南京土壤研究所),南京,210008;Institute of Soil Ecology,GSF-German National Research Center for Environment and Health,85764,Neuherberg Germany 2. Institute of Soil Ecology, GSF-German National Research Center for Environment and Health, 85764 Neuherberg, Germany 3. 土壤与农业可持续发展国家重点实验室,(中国科学院南京土壤研究所),南京,210008 |
| 基金项目： | 国家自然科学基金项目（20707028）、江苏省自然科学基金项目（BK2007264）、中国科学院海外合作伙伴计划项目（CXTD-Z2005-4）和留学人员科技活动择优资助项目资助 |
| 摘 要： | 根据博德特氏菌(Bordetellasp.)的16S rRNA基因序列,设计荧光原位杂交(FISH)检测博德特氏菌的寡核苷酸探针FW_iso_62和FW_iso_761,在20%～60%甲酰胺均有很强的荧光信号。采用探针FW_iso_62及其竞争探针,结合Nycodenz和DAPI技术,建立定量检测土壤中博德特氏菌的DAPI-FISH方法。该方法可排除土壤颗粒的自动荧光对细菌信号的掩盖,保证图片中有大量微生物供统计分析,还能有效保存微生物的原位信息。应用该方法分析土壤中1,2,4-三氯苯降解菌-博德特氏菌,结果未受氯苯污染的农田土壤中没有检测到博德特氏菌,而氯苯污染土壤中检测到大量的博德特氏菌,每克土壤含3.78×106个。将该污染土壤中分离的博德特氏降解菌及其降解菌群接种至农田土壤中,降解菌的数量均随培养而增加,一个月后分别占DAPI计数的1.7%和3.8%。本研究设计的探针可有效用于复杂环境样品中博德特氏菌的定性与定量检测。 |
| 关 键 词： | 博德特氏菌 寡核苷酸探针 荧光原位杂交 三氯苯降解 |
Development and application of specific probes for bordetella sp. in soil by fluorescence in situ hybridization |
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| Wang Fang,M. Schmi,Jiang Xin and R. Schroll.Development and application of specific probes for bordetella sp. in soil by fluorescence in situ hybridization[J].Acta Pedologica Sinica,2010,47(1):51-57. | |
| Authors: | Wang Fang M Schmi Jiang Xin and R Schroll |
| Institution: | State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences; Institute of Soil Ecology, GSF-German National Research Center for Environment and Health;Institute of Soil Ecology, GSF-German National Research Center for Environment and Health;State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences;Institute of Soil Ecology, GSF-German National Research Center for Environment and Health |
| Abstract: | Novel 16S-rRNA targeted oligonucleotide probes(FW_iso_62 and FW_iso_761) were designed to detect Bordetella sp.by fluorescence in situ hybridization(FISH).Strong fluorescence signals with the probes were observed in 20% ~ 60% formamide concentrations.Quantification of Bordetella sp.in soil was developed with the selected probe of FW_iso_62 and its competitor combined with Nycodenz and DAPI technique.The advantages of the established DAPI-FISH: eliminate autofluorescence of soil particles masking the bacteri... |
| Keywords: | Bordetella sp Oligonucleotide probe Fluorescence in situ hybridization Trichlorobenzene degradation |
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