Bacterial community dynamics during the application of a Myxococcus xanthus-inoculated culture medium used for consolidation of ornamental limestone.
Bacterial community dynamics during the application of a Myxococcus xanthus-inoculated culture medium used for consolidation of ornamental limestone.
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DOI：
10.1007/s00248-010-9661-2
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发表时间：
2010-07
影响因子：
3.6
通讯作者：
Teresa Gonzalez-Munoz, Maria
中科院分区：
文献类型：
--
作者：
Pinar, Guadalupe;Jimenez-Lopez, Concepcion;Sterflinger, Katja;Ettenauer, Joerg;Jroundi, Fadwa;Fernandez-Vivas, Antonia;Teresa Gonzalez-Munoz, Maria
在本研究中，我们在实验室条件下研究了接种了黄粘球菌（Myxococcus xanthus）的培养基对采石场和腐烂的观赏碳酸盐石进行固结前后的细菌群落。在固结处理（30 d）期间，监测接种的黄原分枝杆菌的群落结构动态和流行情况。为此，我们选择了一种将变性梯度凝胶电泳（DGGE）指纹图谱与DGGE和测序技术筛选真菌性16S rDNA克隆文库结合的分子策略。利用本研究设计的黄芽孢杆菌特异性引物，采用实时荧光定量PCR （qPCR）对接种菌株进行定量。DGGE和测序分析的结果表明，无论石头的来源如何，同样的产碳微生物被黄豆状芽孢杆菌培养物激活。这些微生物是假单胞菌、芽孢杆菌和短芽孢杆菌。在时间过程实验中，对处理过的结石培养基中的黄原分枝杆菌的监测结果因应用技术的不同而不同。通过依赖培养的方法，只能在治疗的第一天检测到这种细菌，这表明了这些传统技术的局限性。通过PCR-DGGE分析，在实验前3-6天检测到黄原菌。此时，qPCR分析显示，培养基中该细菌的数量在108-106个细胞ml−1之间变化。此后，DGGE分析表明不适合在混合培养中检测黄支霉。然而，使用特异性引物对M. xanthus进行qPCR分析是一种更敏感的检测该细菌的技术，在固结处理结束时，在两种处理过的结石的培养基中发现104 ml - 1的细胞群。本研究提出的分子策略是一种有效的监测系统，用于评估细菌诱导的碳酸盐矿化作为观赏石修复/保护处理的影响。
In this study, we investigated under laboratory conditions the bacterial communities inhabiting quarry and decayed ornamental carbonate stones before and after the application of a Myxococcus xanthus-inoculated culture medium used for consolidation of the stones. The dynamics of the community structure and the prevalence of the inoculated bacterium, M. xanthus, were monitored during the time course of the consolidation treatment (30 days). For this purpose, we selected a molecular strategy combining fingerprinting by denaturing gradient gel electrophoresis (DGGE) with the screening of eubacterial 16S rDNA clone libraries by DGGE and sequencing. Quantification of the inoculated strain was performed by quantitative real-time PCR (qPCR) using M. xanthus-specific primers designed in this work. Results derived from DGGE and sequencing analysis showed that, irrespective of the origin of the stone, the same carbonatogenic microorganisms were activated by the application of a M. xanthus culture. Those microorganisms were Pseudomonas sp., Bacillus sp., and Brevibacillus sp. The monitoring of M. xanthus in the culture media of treated stones during the time course experiment showed disparate results depending on the applied technique. By culture-dependent methods, the detection of this bacterium was only possible in the first day of the treatment, showing the limitation of these conventional techniques. By PCR-DGGE analysis, M. xanthus was detected during the first 3–6 days of the experiment. At this time, the population of this bacterium in the culture media varied between 108–106 cells ml−1, as showed by qPCR analyses. Thereafter, DGGE analyses showed to be not suitable for the detection of M. xanthus in a mixed culture. Nevertheless, qPCR analysis using specific primers for M. xanthus showed to be a more sensitive technique for the detection of this bacterium, revealing a population of 104 cells ml−1 in the culture media of both treated stones at the end of the consolidation treatment. The molecular strategy used in this study is proposed as an effective monitoring system to evaluate the impact of the application of a bacterially induced carbonate mineralization as restoration/conservation treatment for ornamental stones.
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影响因子：
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作者：
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通讯作者：
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影响因子：
3.2
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