【目的】筛选二斑叶螨(Tetranychus urticae)敏感(SS)和抗甲氰菊酯品系(Fe-R)中合适的内参基因,并通过基因相对表达定量PCR研究二斑叶螨抗性品系解毒酶基因表达水平的变化。【方法】采用实时荧光定量PCR(quantitative real time PCR,RT-...【目的】筛选二斑叶螨(Tetranychus urticae)敏感(SS)和抗甲氰菊酯品系(Fe-R)中合适的内参基因,并通过基因相对表达定量PCR研究二斑叶螨抗性品系解毒酶基因表达水平的变化。【方法】采用实时荧光定量PCR(quantitative real time PCR,RT-qPCR)技术,分析二斑叶螨5.8S rRNA、α-tubulin、β-actin、ELF、GAPDH、RPL13a、SDHA和TBP共8个候选内参基因mRNA表达水平的稳定性,并分析二斑叶螨酯酶(Carboxyl/cholinesterases,CCEs)、谷胱甘肽转移酶(Glutathione s-transferase,GSTs)及细胞色素P450单加氧酶(Cytocheome P450 monooxygenases,P450s)等主要解毒酶基因相对表达量。用GeNorm和NormFinder软件进行内参基因稳定性评估,通过比较Ct值的方法进行基因相对表达量计算。【结果】8个内参基因中稳定性最高的为α-tubulin;以α-tubulin为内参基因,二斑叶螨Fe-R品系GSTs基因TuGSTo1、TuGSTd1及P450s基因CYP3D2相对表达量均显著高于SS品系,而CCEs基因TuCCE1的表达量有所降低。【结论】在二斑叶螨SS和Fe-R品系中α-tubulin为理想的内参基因,RT-qPCR研究表明TuGSTo1、TuGSTd1和CYP3D2相对表达量的升高可能是二斑叶螨对甲氰菊酯产生高抗性的分子机理。展开更多
The effects of surfactant Tween-80 on the growth, sulfur oxidation, and expression of selected typical sulfur metabolism relevant genes of Acidithiobacillus ferrooxidans ATCC 23270 were investigated. The results showe...The effects of surfactant Tween-80 on the growth, sulfur oxidation, and expression of selected typical sulfur metabolism relevant genes of Acidithiobacillus ferrooxidans ATCC 23270 were investigated. The results showed that in the presence of 10-2 g/L Tween-80, the growth of A. ferrooxidans and its metabolism on the insoluble substrate S0 and CuFeS2 were promoted. After 24 d of bioleaching, the copper extraction yield of chalcopyrite at 10-2 g/L Tween-80 increased by 16% compared with the bioleaching experiment without Tween-80. FT-IR spectra analysis revealed that the result was probably caused by the extracellular polymeric substances whose composition could be changed by the surfactant addition. RT-qPCR was used to analyze the differential expressions of 17 selected sulfur metabolism relevant genes in response to the addition of Tween-80. Down-regulation of the extracellular protein genes indicated the influence of Tween-80 on bacteria-sulfur adsorption. Variation of the expression level of the enzymes provided a supplement to sulfur metabolism investigation.展开更多
文摘【目的】筛选二斑叶螨(Tetranychus urticae)敏感(SS)和抗甲氰菊酯品系(Fe-R)中合适的内参基因,并通过基因相对表达定量PCR研究二斑叶螨抗性品系解毒酶基因表达水平的变化。【方法】采用实时荧光定量PCR(quantitative real time PCR,RT-qPCR)技术,分析二斑叶螨5.8S rRNA、α-tubulin、β-actin、ELF、GAPDH、RPL13a、SDHA和TBP共8个候选内参基因mRNA表达水平的稳定性,并分析二斑叶螨酯酶(Carboxyl/cholinesterases,CCEs)、谷胱甘肽转移酶(Glutathione s-transferase,GSTs)及细胞色素P450单加氧酶(Cytocheome P450 monooxygenases,P450s)等主要解毒酶基因相对表达量。用GeNorm和NormFinder软件进行内参基因稳定性评估,通过比较Ct值的方法进行基因相对表达量计算。【结果】8个内参基因中稳定性最高的为α-tubulin;以α-tubulin为内参基因,二斑叶螨Fe-R品系GSTs基因TuGSTo1、TuGSTd1及P450s基因CYP3D2相对表达量均显著高于SS品系,而CCEs基因TuCCE1的表达量有所降低。【结论】在二斑叶螨SS和Fe-R品系中α-tubulin为理想的内参基因,RT-qPCR研究表明TuGSTo1、TuGSTd1和CYP3D2相对表达量的升高可能是二斑叶螨对甲氰菊酯产生高抗性的分子机理。
基金Projects (50974140, 51274257) supported by the National Natural Science Foundation of ChinaProject (20090162110054) supported by the PhD Programs Foundation of Ministry of Education of China
文摘The effects of surfactant Tween-80 on the growth, sulfur oxidation, and expression of selected typical sulfur metabolism relevant genes of Acidithiobacillus ferrooxidans ATCC 23270 were investigated. The results showed that in the presence of 10-2 g/L Tween-80, the growth of A. ferrooxidans and its metabolism on the insoluble substrate S0 and CuFeS2 were promoted. After 24 d of bioleaching, the copper extraction yield of chalcopyrite at 10-2 g/L Tween-80 increased by 16% compared with the bioleaching experiment without Tween-80. FT-IR spectra analysis revealed that the result was probably caused by the extracellular polymeric substances whose composition could be changed by the surfactant addition. RT-qPCR was used to analyze the differential expressions of 17 selected sulfur metabolism relevant genes in response to the addition of Tween-80. Down-regulation of the extracellular protein genes indicated the influence of Tween-80 on bacteria-sulfur adsorption. Variation of the expression level of the enzymes provided a supplement to sulfur metabolism investigation.
