Schmidt Silvia, Blom Judith F, Pernthaler Jakob, Berg Gabriele, Baldwin Adam, Mahenthiralingam Eshwar, Eberl Leo
Department of Microbiology, University of Zurich, Zurich, Switzerland.
Environ Microbiol. 2009 Jun;11(6):1422-37. doi: 10.1111/j.1462-2920.2009.01870.x. Epub 2009 Feb 10.
Members of the genus Burkholderia are known for their ability to suppress soil-borne fungal pathogens by the production of various antibiotic compounds. In this study we investigated the role of N-acylhomoserine lactone (AHL)-dependent quorum sensing (QS) in the expression of antifungal traits. Using a quorum quenching approach, that is, by heterologous expression of the Bacillus sp. AiiA lactonase, we show that expression of antifungal activities is AHL-dependent in the large majority of the investigated strains belonging to various Burkholderia species. We demonstrate that in certain strains of Burkholderia ambifaria, Burkholderia pyrrocinia and Burkholderia lata, one of the QS-regulated antifungal agents is pyrrolnitrin (prn), a common broad-spectrum antibiotic that is also produced by some Pseudomonas and Serratia species. To investigate the underlying molecular mechanisms of AHL-dependent prn production in better detail, we inactivated the AHL synthase cepI as well as cepR, which encodes the cognate AHL receptor protein, in B. lata 383. Both QS mutants no longer produced prn as assessed by gas chromatography-mass spectrometry analysis and as a consequence were unable to inhibit growth of Rhizoctonia solani. Using fusions of the lacZ gene to the promoter of the prnABCD operon, which directs the synthesis of prn, we demonstrate that expression of prn is positively regulated by CepR at the level of transcription.
伯克霍尔德氏菌属的成员以其通过产生各种抗生素化合物来抑制土壤传播的真菌病原体的能力而闻名。在本研究中,我们调查了N-酰基高丝氨酸内酯(AHL)依赖性群体感应(QS)在抗真菌特性表达中的作用。使用群体淬灭方法,即通过芽孢杆菌属AiiA内酯酶的异源表达,我们表明在属于各种伯克霍尔德氏菌物种的绝大多数研究菌株中,抗真菌活性的表达是AHL依赖性的。我们证明,在某些双形伯克霍尔德氏菌、吡咯伯克霍尔德氏菌和拉塔伯克霍尔德氏菌菌株中,一种受QS调控的抗真菌剂是硝吡咯菌素(prn),这是一种常见的广谱抗生素,一些假单胞菌属和沙雷氏菌属物种也会产生。为了更详细地研究AHL依赖性prn产生的潜在分子机制,我们在拉塔伯克霍尔德氏菌383中使AHL合酶cepI以及编码同源AHL受体蛋白的cepR失活。通过气相色谱-质谱分析评估,两个QS突变体都不再产生prn,因此无法抑制立枯丝核菌的生长。使用lacZ基因与指导prn合成的prnABCD操纵子启动子的融合体,我们证明prn的表达在转录水平上受到CepR的正调控。