过表达CsMADSs拟南芥的表型变化及CsMADSs表达水平

为探究黄瓜CsMADSs的功能,构建了黄瓜CsMADS08及CsMADS21基因的真核表达载体并转化烟草,研究其在烟草中的亚细胞定位;同时通过农杆菌介导法转化拟南芥,经潮霉素筛选及分子检测,获得阳性株,观察转基因后代植株的表型变化情况;实时荧光定量PCR检测CsMADS08和CsMADS21在转基因后代植株各器官中的表达情况。结果显示,CsMADS08定位于细胞膜和细胞核上,而CsMADS21定位于细胞膜上。表型分析结果显示,过表达CsMADS08基因的拟南芥叶片发紫,侧枝少,且侧枝上出现莲座状的茎生叶;而过表达CsMADS21植株的侧生花序及果荚都比野生型多,且花期早,果实成熟早。荧光定量PCR结果显示,CsMADS08基因主要在茎中表达,而CsMADS21基因则主要在花中有高表达。该研究可为进一步明确黄瓜CsMADSs基因的功能及培育黄瓜新品种提供参考依据。

Study on phenotypic changes and expression of CsMADSs in over-expressing Arabidopsis plants

To investigate the CsMADSs genes which have influence on the growth and development of Arabidopsis thaliana, the eukaryotic expression vectors of CsMADS08 and CsMADS21 genes were constructed and transformed into tobacco to study their subcellular localization. At the same time, the eukaryotic expression vectors were transformed into Arabidopsis through Agrobacterium, and the positive transgenic plants were obtained. Meanwhile, phenotypic changes were observed in transgenic plants, and the expression of CsMADS08 and CsMADS21 were detected by real-time fluorescence quantitative PCR. The results showed that CsMADS08 was positioned on cell membrane and nucleus, while CsMADS21 was positioned on the cell membrane. In addition, purple leaves, fewer branches, and lotus leaves were detected on the lateral branch in the CsMADS08 transgenic plants. On the other hand, there were much more lateral inflorescences and pods, and also earlier flowering and fruits in the CsMADS21 transgenic plants than those of the wild plants. The fluorescent quantitative PCR results showed that CsMADS08 gene was expressed in the main stem, while CsMADS21 gene was mainly expressed in the flowers. The results provided a reference basis for further studying the function of CsMADSs genes and breeding new varieties of cucumber.