Cultivation-independent screening revealed hot spots of IncP-1, IncP-7 and IncP-9 plasmid occurrence in different environmental habitats.
Cultivation-independent screening revealed hot spots of IncP-1, IncP-7 and IncP-9 plasmid occurrence in different environmental habitats.
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DOI：
10.1371/journal.pone.0089922
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Smalla K
中科院分区：
文献类型：
--
作者：
Dealtry S;Ding GC;Weichelt V;Dunon V;Schlüter A;Martini MC;Del Papa MF;Lagares A;Amos GC;Wellington EM;Gaze WH;Sipkema D;Sjöling S;Springael D;Heuer H;van Elsas JD;Thomas C;Smalla K
IncP-1、IncP-7和IncP-9质粒通常携带编码参与降解人造和天然污染物的酶的基因，从而有助于细菌在污染环境中存活。然而，缺乏合适的分子工具往往限制了这些质粒在环境中的检测。在这项研究中，PCR，然后Southern印迹杂交检测质粒特异性序列的存在下，总社区（TC-）DNA或fosmid DNA来自不同的环境和地理区域的样品。开发了一种靶向IncP-9质粒的新型引物系统，并与已建立的IncP-1和IncP-7引物一起沿着应用。筛选TC-DNA从生物纯化系统（BPS），这是用于在农场的农药污染的水的净化揭示了高丰度的IncP-1质粒属于不同的亚群，以及IncP-7和IncP-9。针对9个IncP-9亚组的rep基因的新型IncP-9引物系统允许在具有不同污染源的环境中检测高多样性的IncP-9质粒特异性序列。因此，被污染的位点是潜在携带分解代谢基因的质粒的“热点”。
IncP-1, IncP-7 and IncP-9 plasmids often carry genes encoding enzymes involved in the degradation of man-made and natural contaminants, thus contributing to bacterial survival in polluted environments. However, the lack of suitable molecular tools often limits the detection of these plasmids in the environment. In this study, PCR followed by Southern blot hybridization detected the presence of plasmid-specific sequences in total community (TC-) DNA or fosmid DNA from samples originating from different environments and geographic regions. A novel primer system targeting IncP-9 plasmids was developed and applied along with established primers for IncP-1 and IncP-7. Screening TC-DNA from biopurification systems (BPS) which are used on farms for the purification of pesticide-contaminated water revealed high abundances of IncP-1 plasmids belonging to different subgroups as well as IncP-7 and IncP-9. The novel IncP-9 primer-system targeting the rep gene of nine IncP-9 subgroups allowed the detection of a high diversity of IncP-9 plasmid specific sequences in environments with different sources of pollution. Thus polluted sites are “hot spots” of plasmids potentially carrying catabolic genes.
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