阳春砂DXR超表达提高转基因烟草DXR活性及光合色素含量

Study on Amomum villosum Lour.DXR Overexpression in Enhancing DXR Activity and Photosynthetic Pigment Contents in Transgenic Tobacco

【目的】筛选阳春砂1-去氧-D-木酮糖-5-磷酸还原异构酶(DXR)(AvDXR)转基因烟草T1代植株,鉴定AvDXR基因在模式植物烟草中的功能。【方法】采用抗生素、聚合酶链反应(PCR)法筛选转基因烟草T1代植株;采用半定量逆转录—聚合酶链反应(RT-PCR)法分析目的基因在T1代转基因烟草中的表达情况,采用分光光度法测定酶活性和光合色素含量。【结果】经过抗生素筛选和PCR检测获得AvDXR基因稳定遗传的转基因烟草T1代;AvDXR在部分T1代植株中得到超表达;经测定超表达植株中的DXR活性和光合色素含量均有提高。【结论】AvDXR超表达提高了转基因烟草DXR活性和光合色素含量,获得的AvDXR超表达植株可作为进一步分析萜类成分的材料。

Objective To screen T1 transgenic tobacco plants with Amomum villosum（Av） 1-deoxy-D-xylulose-5-phosphate reductoisomerase（DXR）,and to analyze the biological function of AvDXR in the transgenic tobacco plants.Methods Transgenic tobacco plants were screened by kanamycin and polymerase chain reaction（PCR）.Semi-quantitative reverse transcriptase PCR（RT-PCR） was used to analyze the expression level of AvDXR in transgenic tobacco plants.DXR activity,and chlorophylls and carotenoids contents were determined by spectrometer.Results AvDXR had been steadily inherited and overexpressed in T1 transgenic tobacco plants,and the DXR activity,and chlorophylls and carotenoids contents had been increased.Conclusion Overexpression of AvDXR can enhance DXR activity and photosynthetic pigment contents in transgenic tobacco plants,and the plants with overexpression of AvDXR can be used as the material for further analysis of teperiod components.