Novel assay to measure the plasmid mobilizing potential of mixed microbial communities.
Novel assay to measure the plasmid mobilizing potential of mixed microbial communities.
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DOI：
10.3389/fmicb.2014.00730
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发表时间：
2014
影响因子：
5.2
通讯作者：
Smets BF
中科院分区：
文献类型：
--
作者：
Klümper U;Droumpali A;Dechesne A;Smets BF
可移动质粒缺乏完全接合所需的基因，因此不具有自传输性。相反，它们依靠偶联质粒的偶联系统水平转移到新的受体。虽然已经研究了群落容许度，即混合微生物群落中可以接受自传播的偶联质粒的比例，但尚未探索群落动员缺乏偶联系统的质粒的内在能力。在这里，我们提出了一个新的框架和实验方法来估计混合社区的动员潜力。我们比较了一个可动员质粒的转移频率，以动员和偶联质粒为模型菌株和被测社区测量。以携带gfp标记的可动员IncP-1α质粒RSF1010的恶臭假单胞菌为供体菌株，与携带可动员IncP-1α质粒RP4的恶臭假单胞菌或从家用淋浴管内壁提取的模型细菌群落进行固体表面交配实验。此外，我们估计了同一群落对以恶臭杆菌为供体菌株的RP4的容忍度。模型群落对RP4的容忍度[每受体1.16 × 10-4个转偶联物（T/R）]与之前测量的土壤微生物群落相似。RSF1010被模型群落动员的频率为1.16 × 10-5 T/R，仅比其对RP4的容许度低一个数量级。考虑到(i)动员需要在受体的允许部分内存在动员的共轭质粒，这种动员频率出乎意料地高；（ii）在恶臭假单胞菌纯培养实验中，只有大约一半的供体在第一步中接受了偶联质粒，通过RP4对RSF1010进行了再动员。需要进一步的工作来确定质粒动员潜力如何在微生物群落内部和不同的微生物群落中变化。这种方法有可能提供这样的见解；此外，它允许直接分离原位动员质粒及其内源性宿主。
Mobilizable plasmids lack necessary genes for complete conjugation and are therefore non-self-transmissible. Instead, they rely on the conjugation system of conjugal plasmids to be horizontally transferred to new recipients. While community permissiveness, the fraction of a mixed microbial community that can receive self-transmissible conjugal plasmids, has been studied, the intrinsic ability of a community to mobilize plasmids that lack conjugation systems is unexplored. Here, we present a novel framework and experimental method to estimate the mobilization potential of mixed communities. We compare the transfer frequency of a mobilizable plasmid to that of a mobilizing and conjugal plasmid measured for a model strain and for the assayed community. With Pseudomonas putida carrying the gfp-tagged mobilizable IncQ plasmid RSF1010 as donor strain, we conducted solid surface mating experiments with either a P. putida strain carrying the mobilizing IncP-1α plasmid RP4 or a model bacterial community that was extracted from the inner walls of a domestic shower conduit. Additionally, we estimated the permissiveness of the same community for RP4 using P. putida as donor strain. The permissiveness of the model community for RP4 [at 1.16 × 10-4 transconjugants per recipient (T/R)] was similar to that previously measured for soil microbial communities. RSF1010 was mobilized by the model community at a frequency of 1.16 × 10-5 T/R, only one order of magnitude lower than its permissiveness to RP4. This mobilization frequency is unexpectedly high considering that (i) mobilization requires the presence of mobilizing conjugal plasmids within the permissive fraction of the recipients; (ii) in pure culture experiments with P. putida retromobilization of RSF1010 through RP4 only took place in approximately half of the donors receiving the conjugal plasmid in the first step. Further work is needed to establish how plasmid mobilization potential varies within and across microbial communities. This method has the potential to provide such insights; in addition it allows for the direct isolation of in situ mobilizing plasmids together with their endogenous hosts.
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影响因子：
4.4
作者：
HILL, KE;WEIGHTMAN, AJ;FRY, JC
通讯作者：
FRY, JC
广泛的宿主范围INCQ质粒的复制和共轭动员。
影响因子：
2.6
作者：
Meyer R
通讯作者：
Meyer R
影响因子：
4.4
作者：
REASONER, DJ;BLANNON, JC;GELDREICH, EE
通讯作者：
GELDREICH, EE
DOI：
10.1007/978-1-4419-7931-5_7
发表时间：
2011-01-01
期刊：
MICROBES AND MICROBIAL TECHNOLOGY: AGRICULTURAL AND ENVIRONMENTAL APPLICATIONS
影响因子：
--
作者：
Grohmann, Elisabeth
通讯作者：
Grohmann, Elisabeth
影响因子：
2.6
作者：
Król JE;Wojtowicz AJ;Rogers LM;Heuer H;Smalla K;Krone SM;Top EM
通讯作者：
Top EM