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Functional identification of phenazine biosynthesis genes in plant pathogenic bacteria Pseudomonas syringae pv. tomato and Xanthomonas oryzae pv. oryzae 被引量:1
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作者 LI Wen XU You-ping +4 位作者 jean-pierre munyampundu XU Xin QI Xian-fei GU Yuan CAI Xin-zhong 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2016年第4期812-821,共10页
Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens. In this study, we identified phenazine biosynthesis (phz) genes in two ge... Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens. In this study, we identified phenazine biosynthesis (phz) genes in two genome-completed plant pathogenic bacteria Pseudomonas syringae pv. tomato (Pst) DC3000 and Xanthomonas oryzae pv. oryzae (Xoo) PXO99A. Unlike the phz genes in typical phenazine-producing pseudomonads, phz homologs in Pst DC3000 and Xoo PXO99A consisted of phzC/D/E/F/G and phzC/E1/E2/F/G, respectively, and the both were not organized into an operon. Detection experiments demonstrated that phenazine-l-carboxylic acid (PCA) of Pst DC3000 accumulated to 13.4 IJg L-1, while that of Xoo PXO99A was almost undetectable. Moreover, Pst DC3000 was resistant to 1 mg mL-1 PCA, while Xoo PXO99A was sensitive to 50 IJg mL ~ PCA. Furthermore, mutation of phzF blocked the PCA production and significantly reduced the pathogenicity of Pst DC3000 in tomato, while the complementary strains restored these phenotypes. These results revealed that Pst DC3000 produces low level of and is resistant to phenazines and thus is unable to be biologically controlled by phenazines. Additionally, phz-mediated PCA production is required for full pathogenicity of Pst DC3000. To our knowledge, this is the first report of PCA production and its function in pathogenicity of a plant pathogenic P. syringae strain. 展开更多
关键词 PATHOGENICITY phenazine biosynthesis genes phenazine-l-carboxylic acid plant pathogenic bacteria Pseudomonas syringae pv. tomato Xanthomonas oryzae pv. oryzae
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问号钩端螺旋体过氧化物还原酶AhpC在氧化应激中的功能研究 被引量:2
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作者 罗依惠 吴亦斐 jean-pierre munyampundu 《中华微生物学和免疫学杂志》 CAS CSCD 北大核心 2018年第3期193-198,共6页
目的确定问号钩端螺旋体(简称钩体)赖株AhpC蛋白的过氧化物还原酶活性及其在问号钩体感染宿主细胞过程中是否具有抵抗氧化应激的功能。 方法构建问号钩体黄疸出血群赖型赖株ahpC基因原核表达系统。Ni-NTA亲和层析法提纯目的重组蛋白r... 目的确定问号钩端螺旋体(简称钩体)赖株AhpC蛋白的过氧化物还原酶活性及其在问号钩体感染宿主细胞过程中是否具有抵抗氧化应激的功能。 方法构建问号钩体黄疸出血群赖型赖株ahpC基因原核表达系统。Ni-NTA亲和层析法提纯目的重组蛋白rAhpC,检测其酶学活性和保护DNA不被氧化的功能。用定点突变方法验证AhpC的过氧化半胱氨酸和还原性半胱氨酸。经不同浓度的Conoidin A抑制问号钩体过氧化物还原酶活性后,比较抑制前后构体内的活性氧自由基(reactive oxygen species,ROS)水平和钩体存活率的变化。结果所构建的原核表达系统能有效表达rAhpC。rAhpC具有过氧化物还原酶的活性,其催化反应依赖于硫氧还蛋白和硫氧还蛋白还原酶系统。AhpC分子中含有的两个半胱氨酸对维持酶的活性起着至关重要的作用,其中Cys47是AhpC的过氧化半胱氨酸,而Cys167是还原性半胱氨酸。AhpC能够保护质粒DNA免遭过氧化氢的氧化损伤。经不同浓度的Conoidin A抑制问号钩体的过氧化物还原酶的活性后,钩体在巨噬细胞内的存活率明显下降,并且呈现出剂量依赖的效应,表明钩体存活率的降低与钩体的过氧化物还原酶活性丧失从而无法有效降解构体内活性氧水平有着密切的关系。结论问号钩体的AhpC具有过氧化物还原酶的活性,能抵抗宿主细胞引起的氧化损伤,与钩体在巨噬细胞内的存活有关。 展开更多
关键词 问号钩端螺旋体 ahpC基因 氧化应激 存活
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