Biochemical, genetic, and zoosporicidal properties of cyclic lipopeptide surfactants produced by Pseudomonas fluorescens

被引:151
作者
de Souza, JT
de Boer, M
de Waard, P
van Beek, TA
Raaijmakers, JM
机构
[1] Univ Wageningen & Res Ctr, Dept Plant Sci, Phytopathol Lab, NL-6709 PD Wageningen, Netherlands
[2] Sect Flower Bulbs, Appl Plant Res, NL-2160 AB Lisse, Netherlands
[3] Wageningen Univ, NMR Ctr, NL-6700 ET Wageningen, Netherlands
[4] Univ Wageningen & Res Ctr, Organ Chem Lab, Nat Prod Chem Grp, NL-6703 HB Wageningen, Netherlands
关键词
D O I
10.1128/AEM.69.12.7161-7172.2003
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Zoospores play an important role in the infection of plant and animal hosts by oomycetes and other zoosporic fungi. In this study, six fluorescent Pseudomonas isolates with zoosporicidal activities were obtained from the wheat rhizosphere. Zoospores of multiple oomycetes, including Pythium species, Albugo candida, and Phytophthora infestans, were rendered immotile within 30 s of exposure to cell suspensions or cell culture supernatants of the six isolates, and subsequent lysis occurred within 60 s. The representative strain SS101, identified as Pseudomonas fluorescens biovar II, reduced the surface tension of water from 73 to 30 mN m(-1). The application of cell suspensions of strain SS101 to soil or hyacinth bulbs provided significant protection against root rot caused by Pythium intermedium. Five Tn5 mutants of strain SS101 lacked the abilities to reduce the surface tension of water and to cause lysis of zoospores. Genetic characterization of two surfactant-deficient mutants showed that the transposons had integrated into condensation domains of peptide synthetases. A partially purified extract from strain SS101 reduced the surface tension of water to 30 mN m(-1) and reached the critical micelle concentration at 25 mug ml(-1). Reverse-phase high-performance liquid chromatography yielded eight different fractions, five of which had surface activity and caused lysis of zoospores. Mass spectrometry and nuclear magnetic resonance analyses allowed the identification of the main constituent as a cyclic lipopeptide (1,139 Da) containing nine amino acids and a 10-carbon hydroxy fatty acid. The other four zoosporicidal fractions were closely related to the main constituent, with molecular masses ranging from 1,111 to 1,169 Da.
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页码:7161 / 7172
页数:12
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