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Nonribosomal Peptides, Key Biocontrol Components for Pseudomonas fluorescens In5, Isolated from a Greenlandic Suppressive Soil

机译:非核糖体肽,从格陵兰抑制土壤中分离出来的荧光假单胞菌 In5的关键生物防治成分

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ABSTRACT Potatoes are cultivated in southwest Greenland without the use of pesticides and with limited crop rotation. Despite the fact that plant-pathogenic fungi are present, no severe-disease outbreaks have yet been observed. In this report, we document that a potato soil at Inneruulalik in southern Greenland is suppressive against Rhizoctonia solani Ag3 and uncover the suppressive antifungal mechanism of a highly potent biocontrol bacterium, Pseudomonas fluorescens In5, isolated from the suppressive potato soil. A combination of molecular genetics, genomics, and matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) imaging mass spectrometry (IMS) revealed an antifungal genomic island in P.?fluorescens In5 encoding two nonribosomal peptides, nunamycin and nunapeptin, which are key components for the biocontrol activity by strain In5 in vitro and in soil microcosm experiments. Furthermore, complex microbial behaviors were highlighted. Whereas nunamycin was demonstrated to inhibit the mycelial growth of R.?solani Ag3, but not that of Pythium aphanidermatum , nunapeptin instead inhibited P.?aphanidermatum but not R.?solani Ag3. Moreover, the synthesis of nunamycin by P.?fluorescens In5 was inhibited in the presence of P.?aphanidermatum . Further characterization of the two peptides revealed nunamycin to be a monochlorinated 9-amino-acid cyclic lipopeptide with similarity to members of the syringomycin group, whereas nunapeptin was a 22-amino-acid cyclic lipopeptide with similarity to corpeptin and syringopeptin. IMPORTANCE Crop rotation and systematic pest management are used to only a limited extent in Greenlandic potato farming. Nonetheless, although plant-pathogenic fungi are present in the soil, the farmers do not experience major plant disease outbreaks. Here, we show that a Greenlandic potato soil is suppressive against Rhizoctonia solani , and we unravel the key biocontrol components for Pseudomonas fluorescens In5, one of the potent biocontrol bacteria isolated from this Greenlandic suppressive soil. Using a combination of molecular genetics, genomics, and microbial imaging mass spectrometry, we show that two cyclic lipopeptides, nunamycin and nunapeptin, are important for the biocontrol activity of P.?fluorescens In5 both in vitro and in microcosm assays. Furthermore, we demonstrate that the synthesis of nunamycin is repressed by the oomycete Pythium aphanidermatum . Overall, our report provides important insight into interkingdom interference between bacteria and fungi/oomycetes.
机译:摘要马铃薯是在格陵兰西南部种植的,无需使用杀虫剂且轮作受限。尽管存在植物致病真菌,但尚未观察到严重的疾病暴发。在此报告中,我们记录了格陵兰岛南部Inneruulalik的马铃薯土壤对soliz Rhizoctonia solani Ag3的抑制作用,并揭示了从抑制性马铃薯土壤中分离出的高效生物防治细菌Pseudomonas fluorescens In5的抑制性抗真菌机制。分子遗传学,基因组学和基质辅助激光解吸电离飞行时间(MALDI-TOF)成像质谱(IMS)的结合揭示了在P.?fluorescens In5中有一个抗真菌基因组岛,该岛上编码两种非核糖体肽,努那霉素和努那肽,它们是In5菌株在体外和土壤微观实验中的生物防治活性的关键成分。此外,强调了复杂的微生物行为。努纳霉素被证明可以抑制R.?solani Ag3的菌丝体生长,但不能抑制瓜果腐霉的腐霉菌,但是,Nunapeptin却可以抑制R.?solani Ag3的菌丝生长。此外,在P.αaphanidermatum的存在下,P.α荧光假单胞菌In5合成了纳那霉素。两种多肽的进一步表征显示,纳那霉素是一种单丁酰化的9个氨基酸的环状脂肽,与丁香霉素基团的成员相似,而那肽素则是一种22个氨基酸的环状脂肽,与蝎蛋白和丁香脂素相似。重要信息在格陵兰马铃薯种植中仅有限地使用了作物轮作和系统的病虫害管理。尽管如此,尽管土壤中存在植物病原性真菌,但农民并未经历重大的植物病暴发。在这里,我们显示了格陵兰马铃薯土壤对solani soliz的抑制作用,并且我们揭示了荧光假单胞菌In5的关键生物防治成分,Pseudomonas fluorescens In5是从这种格陵兰抑制土壤中分离的有效生物防治细菌之一。结合使用分子遗传学,基因组学和微生物成像质谱技术,我们表明两个环脂肽,努纳霉素和nunapeptin,对于荧光假单胞菌In5在体外和微观检测中的生物防治活性都很重要。此外,我们证明努那霉素的合成受卵菌腐霉腐霉的抑制。总体而言,我们的报告对细菌与真菌/卵菌之间的相互干扰提供了重要的见解。

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