中文摘要：

目的构建烟曲霉额外拷贝菌株，了解额外拷贝烟曲霉sho1、pbs2基因能否增强菌株对高渗透压、过氧化氢（H：O2）、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sho1、pbs2基因的额外拷贝菌株，采用Real-time PCR方法检测额外拷贝株中sho1、pbs2的表达情况。观察并比较缺陷株、额外拷贝株对NaCl（1mol／L）、H2O2（5mmol／L）、刚果红（400mg／L）及碱性pH（10．0）应激的反应。结果获得了含有烟曲霉sho1、pbs2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sho1、pbs2的表达水平增高，对NaCl（1mol／L）、H2O2（5mmol／L）、刚果红（400mg／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△sho1、△pbs2对NaCl（1mol／L）、H2O2（5mmol／L）、碱性pH（10．0）的敏感性高于野生株AF293。△sho1对刚果红（400mg／L）的敏感性高于野生株，△pbs2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sho1或pbs2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。

英文摘要：

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 MCshol or MCpbs2. RT-PCR was used to detect the expression levels ofsho I in MCshol andpbs 2 in MCpbs2 strains. Then strains harboring extra copies as well as sho 1 andpbs 2 deleted mutants were treated with NaCl, H2O2 , Congo red and alkaline pH stresses and the re- sistance phenotypes were observed. Results Mutants harboring extra copies of sho 1 or pbs 2 showed higher expression levels of sho 1 orpbs 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. A 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.