转PcRS基因拟南芥植株表型分析

为了研究虎杖(Polygonum cuspidatum Sieb.et Zucc.)白藜芦醇合酶基因PcRS的功能,以转PcRS基因拟南芥为材料,对其进行分子鉴定和表型研究.利用Southern blot和Northern blot验证了外源PcRS基因在转基因拟南芥基因组中的整合和表达.对经过选育得到的T3代纯合株系(L4、L6、L15)进行性状观察.在体视显微镜下观察野生型和转基因拟南芥植株表型;采用分光光度法测定低氮培养基上的子叶花青素含量.结果显示,与野生型拟南芥相比,转基因拟南芥的种皮颜色发生改变呈浅黄褐色,并且子叶中花青素含量显著减少.这表明PcRS基因可能导致转基因拟南芥的黄酮类物质合成受阻.

Phenotypic analysis of the transgenic PcRS Arabidopsis plants

To screen transgenic Arabidopsis plants with Polygonum cuspidatum resveratrol synthase (PcRS), and to analyze the biological function of PcRS in the transgenic Arabidopsis plants, the integration and expression of transgene in the genome of Arabidopsis was confirmed by Southern blot and Northern blot analyses. With stereomicroscope, the plant phenotypes were observed in transgenic Arabidopsis and wild-type Arabidopsis. Arabidopsis plants were grown on medium devoid of nitrogen sources and anthocyanin content was quantitated by using a spectrophotometer. The homozygous T3 progenies of three transgenic lines (L4, L6 and L15) were obtained. In this paper, we observed that transgenic plants had seeds with a pale fawn colour and accumulated less anthocyanin in cotyledons in contrast to the wild -type plants. Overexpression of PcRS could cause altered seed coat pigmentation and significant reduction in anthocyanin levels. These results suggested that additional PcRS activities lead to the disturbance of the subsequent flavonoid biosynthesis steps in Arabidopsis.