Ge Yi-he, Zhao Yan-hong, Chen Li-juan, Miao Jing, Wen Lu
Department of Biology, Huaiyin Normal University, Huaiyin 223000, China.
Wei Sheng Wu Xue Bao. 2007 Jun;47(3):441-6.
A new bacterium with potential biocontrol ability, Pseudomonas sp. M18, was isolated from the soil of agricultural field in suburb of Shanghai (China). It had been demonstrated that biosynthesis and secretion of phenazine-1-carboxylic acid and pyoluteorin in Pseudomonas sp. M18 contributes to its suppression of soilborne pathogens. In order to study the correlation and regulatory mechanism of two antifungal compounds biosynthesis, the mutant M18T and M18Z1 were constructed with insertion of the gentamycin resistance gene cassette (aacC1), respectively. With introduction of the translational fusion pMEAZ (pltA'-' lacZ) into the wild type strain MI8 or the pit-mutant M18T, respectively, it was found that beta-galactosidase activities of the mutant M18T (pMEAZ) are remarkably enhanced by adding a certain amount of pyoluteorin in KMB medium. The results indicated that pyoluteorin might positively autoinduce expression of the pit gene loci. In investigating the correlation of two antifungal agents, it was showed that the pyoluteorin-negative mutant MI8T produces the same level of phenazine-1-carboxylic acid in comparison with the wild type strain M18. Overexpression of the plt gene loci does not result in decrease of phenazine-1-carboxylic acid in a pltZ-mutant of Pseudomonas sp. M18. However, the distinct decrease of phenazine-1-carboxylic acid biosynthesis does lead to enhanced biosynthesis of pyoluteorin in the mutant M18Z1. Addition of phenazine-1-carboxylic acid in KMB medium makes the mutant M18S produce less pyoluteorin. These results indicated that a special correlation of secondary antifungal agents biosynthesis seems to be existed in Pseudomonas sp. M18, i.e., production of pyoluteorin does not exert any influence on expression of the phz gene cluster, while phenazine-1-carboxylic acid makes negative impact on the biosynthesis of pyoluteorin.
从上海郊区(中国)农田土壤中分离出一种具有潜在生物防治能力的新细菌——假单胞菌属M18。已证明假单胞菌属M18中吩嗪 - 1 - 羧酸和绿脓菌素的生物合成与分泌有助于其对土传病原菌的抑制作用。为了研究两种抗真菌化合物生物合成的相关性及调控机制,分别构建了插入庆大霉素抗性基因盒(aacC1)的突变体M18T和M18Z1。将翻译融合质粒pMEAZ(pltA'-'lacZ)分别导入野生型菌株MI8或pit突变体M18T,发现在KMB培养基中添加一定量的绿脓菌素可显著增强突变体M18T(pMEAZ)的β - 半乳糖苷酶活性。结果表明,绿脓菌素可能正向自诱导pit基因座的表达。在研究两种抗真菌剂的相关性时,发现绿脓菌素阴性突变体MI8T与野生型菌株M18相比,吩嗪 - 1 - 羧酸的产生水平相同。在假单胞菌属M18的pltZ突变体中,plt基因座的过表达不会导致吩嗪 - 1 - 羧酸的减少。然而,吩嗪 - 1 - 羧酸生物合成的明显减少确实导致突变体M18Z1中绿脓菌素生物合成的增强。在KMB培养基中添加吩嗪 - 1 - 羧酸会使突变体M18S产生较少的绿脓菌素。这些结果表明,假单胞菌属M18中似乎存在次级抗真菌剂生物合成的特殊相关性,即绿脓菌素的产生对phz基因簇的表达没有任何影响,而吩嗪 - 1 - 羧酸对绿脓菌素的生物合成有负面影响。