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土壤微型节肢动物白符跳(弹尾目)肠道中大肠杆菌携带的接合型和可移动质粒的属间转移

Intergeneric transfer of conjugative and mobilizable plasmids harbored by Escherichia coli in the gut of the soil microarthropod Folsomia candida (Collembola).

作者信息

Hoffmann A, Thimm T, Dröge M, Moore E R, Munch J C, Tebbe C C

机构信息

Institut für Bodenbiologie, Bundesforschungsanstalt für Landwirtschaft, Braunschweig, Germany.

出版信息

Appl Environ Microbiol. 1998 Jul;64(7):2652-9. doi: 10.1128/AEM.64.7.2652-2659.1998.

Abstract

The gut of the soil microarthropod Folsomia candida provides a habitat for a high density of bacterial cells (T. Thimm, A. Hoffmann, H. Borkott, J. C. Munch, and C. C. Tebbe, Appl. Environ. Microbiol. 64:2660-2669, 1998). We investigated whether these gut bacteria act as recipients for plasmids from Escherichia coli. Filter mating with E. coli donor cells and collected feces of F. candida revealed that the broad-host-range conjugative plasmid pRP4-luc (pRP4 with a luciferase marker gene) transferred to fecal bacteria at estimated frequencies of 5.4 x 10(-1) transconjugants per donor. The mobilizable plasmid pSUP104-luc was transferred from the IncQ mobilizing strain E. coli S17-1 and less efficiently from the IncF1 mobilizing strain NM522 but not from the nonmobilizing strain HB101. When S17-1 donor strains were fed to F. candida, transconjugants of pRP4-luc and pSUP104-luc were isolated from feces. Additionally, the narrow-host-range plasmid pSUP202-luc was transferred to indigenous bacteria, which, however, could not maintain this plasmid. Inhibition experiments with nalidixic acid indicated that pRP4-luc plasmid transfer took place in the gut rather than in the feces. A remarkable diversity of transconjugants was isolated in this study: from a total of 264 transconjugants, 15 strains belonging to the alpha, beta, or gamma subclass of the class Proteobacteria were identified by DNA sequencing of the PCR-amplified 16S rRNA genes and substrate utilization assays (Biolog). Except for Alcaligenes faecalis, which was identified by the Biolog assay, none of the isolates was identical to reference strains from data banks. This study indicates the importance of the microarthropod gut for enhanced conjugative gene transfer in soil microbial communities.

摘要

土壤微型节肢动物白符跳的肠道为高密度的细菌细胞提供了一个栖息地(T. 蒂姆、A. 霍夫曼、H. 博尔科特、J. C. 蒙克和C. C. 特贝,《应用与环境微生物学》64:2660 - 2669,1998年)。我们研究了这些肠道细菌是否作为来自大肠杆菌的质粒的受体。将白符跳与大肠杆菌供体细胞进行滤膜杂交,并收集其粪便,结果表明,广宿主范围的接合性质粒pRP4 - luc(带有荧光素酶标记基因的pRP4)以估计每供体5.4×10⁻¹个转接合子的频率转移到粪便细菌中。可移动质粒pSUP104 - luc从IncQ型可移动菌株大肠杆菌S17 - 1转移,从IncFⅠ型可移动菌株NM522转移效率较低,但从非可移动菌株HB101不发生转移。当将S17 - 1供体菌株投喂给白符跳时,从粪便中分离到了pRP4 - luc和pSUP104 - luc的转接合子。此外,窄宿主范围的质粒pSUP202 - luc转移到了本土细菌中,然而这些细菌无法维持该质粒。用萘啶酸进行的抑制实验表明,pRP4 - luc质粒转移发生在肠道而非粪便中。在本研究中分离到了种类显著多样的转接合子:在总共264个转接合子中,通过对PCR扩增的16S rRNA基因进行DNA测序和底物利用分析(Biolog),鉴定出15个属于变形菌门α、β或γ亚类的菌株。除了通过Biolog分析鉴定出的粪产碱菌外,没有一个分离株与数据库中的参考菌株相同。本研究表明微型节肢动物肠道对于增强土壤微生物群落中的接合基因转移具有重要意义。

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