Wageningen University, Department of Plant Sciences, Laboratory of Phytopathology, Binnenhaven 5, P.O. Box 8025, 6709 PD Wageningen, The Netherlands.
FEMS Microbiol Ecol. 2003 Feb 1;43(1):21-34. doi: 10.1111/j.1574-6941.2003.tb01042.x.
Abstract Pyrrolnitrin (PRN) and pyoluteorin (PLT) are broad-spectrum antibiotics produced by several strains of Pseudomonas and Burkholderia species. Both antibiotics play an important role in the suppression of multiple plant pathogenic fungi. Primers were developed from conserved sequences and amplified prnD and pltC fragments from 18 Pseudomonas and four Burkholderia spp. of worldwide origin that produce either PRN or PLT or both. Subsequent RFLP (restriction fragment length polymorphisms) analysis of the 438-bp pltC fragment showed no polymorphisms among PLT-producing Pseudomonas strains. Polymorphisms within the 786-bp prnD fragment, however, allowed the assessment of the diversity among PRN-producing Pseudomonas and Burkholderia spp. to a level similar to that obtained by three 10-mer primers in random amplified polymorphic DNA analysis. Phylogenetic analysis of 16S rDNA sequences of strains representative of PRN-producing Pseudomonas and Burkholderia species correlated well with their taxonomic status. Phylogenetic relationships inferred from each of the four prn genes and from the complete sequence of the prn biosynthetic locus were similar to 16S rDNA-based phylogeny for most strains, except for Burkholderia pyrrocinia DSM 10685. Both RFLP analysis and comparison of the prn gene sequences showed that B. pyrrocinia DSM 10685 was more closely related to PRN-producing Pseudomonas strains, suggesting that lateral gene transfer may have occurred. Colony hybridization and PCR with PRN- and PLT-specific probes and primers showed that Pseudomonas and Burkholderia spp. harboring the prnD and pltC gene were not present at detectable levels on roots of wheat grown in five agricultural soils collected in The Netherlands, two of them being naturally suppressive to Gaeumannomyces graminis var. tritici. These results suggest that PRN- and PLT-producing Pseudomonas and Burkholderia sp. do not contribute to the natural suppressiveness found in these Dutch take-all decline soils.
硝吡咯菌素（PRN）和吡咯菌素（PLT）是由假单胞菌和伯克霍尔德菌属的几种菌株产生的广谱抗生素。这两种抗生素在抑制多种植物病原真菌方面都发挥了重要作用。从保守序列中设计引物，从来自世界各地的产生 PRN 或 PLT 或两者的 18 种假单胞菌和 4 种伯克霍尔德菌属菌株中扩增 prnD 和 pltC 片段。随后对 438bp 的 pltC 片段进行的 RFLP（限制性片段长度多态性）分析表明，PLT 产生的假单胞菌菌株之间没有多态性。然而，prnD 片段内的多态性允许评估 PRN 产生的假单胞菌和伯克霍尔德菌属的多样性，其水平与随机扩增多态性 DNA 分析中使用的三个 10 -mer 引物获得的水平相似。代表 PRN 产生的假单胞菌和伯克霍尔德菌属的菌株的 16S rDNA 序列的系统发育分析与它们的分类地位密切相关。从四个 prn 基因中的每一个以及从 prn 生物合成基因座的完整序列推断的系统发育关系与大多数菌株的基于 16S rDNA 的系统发育关系相似，除了伯克霍尔德菌属 pyrrocinia DSM 10685 之外。RFLP 分析和 prn 基因序列比较表明，伯克霍尔德菌属 pyrrocinia DSM 10685 与 PRN 产生的假单胞菌菌株更为密切相关，表明可能发生了横向基因转移。用 PRN 和 PLT 特异性探针和引物进行的菌落杂交和 PCR 显示，在荷兰采集的五种农业土壤中生长的小麦根上未检测到携带 prnD 和 pltC 基因的假单胞菌和伯克霍尔德菌属，其中两个对禾谷丝核菌 var. tritici 具有天然抑制作用。这些结果表明，在这些荷兰小麦全蚀病衰退土壤中发现的 PRN 和 PLT 产生的假单胞菌和伯克霍尔德菌属不会导致自然抑制。
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