烟曲霉sho1、pbs2基因额外拷贝对几种应激能力的影响

摘要/Abstract

摘要： 目的构建烟曲霉额外拷贝菌株,了解额外拷贝烟曲霉sho 1、pbs 2基因能否增强菌株对高渗透压、过氧化氢 (H₂O₂)、碱性pH、刚果红应激的抵抗能力,探讨HOG通路 (high osmolarity glycerol pathway)参与的应激反应。方法用原生质体法构建分别含有烟曲霉sho 1、pbs 2基因的额外拷贝菌株,采用Real-time PCR方法检测额外拷贝株中sho 1、pbs 2的表达情况。观察并比较缺陷株、额外拷贝株对NaCl (1 mol/L)、H₂O₂ (5 mmol/L)、刚果红 (400 mg/L)及碱性pH (10.0)应激的反应。结果获得了含有烟曲霉sho 1、pbs 2基因的额外拷贝菌株MCsho1、MCpbs2,和含空白质粒的对照株Empty。额外拷贝株sho 1、pbs 2的表达水平增高,对NaCl (1 mol/L)、H₂O₂ (5 mmol/L)、刚果红 (400 mg/L)、碱性pH (10.0)应激的抵抗强于Empty。MCpbs2对这些应激的抵抗较MCsho1更显著。烟曲霉缺陷株△sho 1、△pbs 2对NaCl (1 mol/L)、H₂O₂ (5 mmol/L)、碱性pH (10.0)的敏感性高于野生株AF293。△sho 1对刚果红 (400 mg/L)的敏感性高于野生株,△pbs 2对刚果红的敏感性与野生株比,无显著差别。结论额外拷贝烟曲霉sho 1或pbs 2基因能增强菌株对高渗透压、氧化压力、刚果红、碱性pH应激的抵抗能力。

关键词: 烟曲霉, sho 1, pbs 2, 渗透应激, 氧化应激, 刚果红, 碱性pH

Abstract: Objective To explore the role of extra copies of sho 1 or pbs 2 in resistance to hyperosmotic stress,oxidative stress, cell wall stress and alkaline pH stress in A. fumigatus.Methods The reconstructed plasmid harboring a sho 1 gene or a pbs 2 gene were transformed into A. fumigatus by using protoplast transformation method and the resulting strains were designated MCsho1 or MCpbs2. RT-PCR was used to detect the expression levels of sho 1 in MCsho1 and pbs 2 in MCpbs2 strains. Then strains harboring extra copies as well as sho 1 and pbs 2 deleted mutants were treated with NaCl, H₂O₂, Congo red and alkaline pH stresses and the resistance phenotypes were observed.Results Mutants harboring extra copies of sho 1 or pbs 2 showed higher expression levels of sho 1 or pbs 2 respectively. Extra copies of sho 1 or pbs 2 conferred resistance to NaCl (1 mol/L), H₂O₂ (5 mmol/L), Congo red (400 mg/L)or alkaline pH (10.0)stress.sho 1 and pbs 2 deleted mutants showed higher sensitivity to NaCl (1 mol/L),H₂O₂ (5 mmol/L)and alkaline pH (10.0)stress than the wild strain. △pbs 2 deleted mutant showed similar sensitivity to Congo red (400mg/L) compared to the wild strain.Conclusion Extra copies of sho 1 or pbs 2 conferred resistance to hyperosmotic stress, oxidative stress, cell wall stress and alkaline pH stress in A. fumigatus.

Key words: Aspergillus fumigatus, sho 1, pbs 2, Hyperosmotic stress, Oxidative stress, Congo red, Alkaline pH

中图分类号:

R379.6

马冬梅, 季雅娟, 阳芳, 刘伟, 万喆, 李若瑜. 烟曲霉sho1、pbs2基因额外拷贝对几种应激能力的影响[J]. 中国真菌学杂志, 2012, (4): 193-198.

MA Dong-mei, JI Ya-juan, YANG Fang, LIU Wei, WAN Zhe, LI Ruo-yu. The effect of extra copies of sho 1 or pbs 2 gene on the adaptive ability of Aspergillus fumigatus to several stresses[J]. Chinese Journal of Mycology, 2012, (4): 193-198.

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