蕙兰CfCIN基因克隆及其功能分析

以蕙兰(Cymbidium faberi Rolfe)为试验材料,采用RT-PCR技术克隆了TCP家族的CIN同源基因,开放阅读框长1 161bp,编码386个氨基酸,将其命名为CfCIN(GenBank登录号为KJ956809)。为进一步分析CfCIN的功能,构建了植物表达载体,采用农杆菌介导法转化非洲紫罗兰叶片,获得了转化植株并对转基因植株进行了性状分析。结果显示:与野生型非洲紫罗兰叶片相比,转基因植株的叶片更大,由圆形变为卵圆形,叶缘由平整光滑变为有缺刻且稍向后卷曲,叶脉明显,叶柄红,花器官形状变化不明显。研究表明,CfCIN可能参与调控植物叶片的形态建成。

Isolation and Functional Verification of CfCIN in Cymbidium faberi

Flower bud of Cymbidium faberi was chosen as experiment material for gene cloning. A CIN like gene of TCP family was amplified through RT PCR technique and named as CfCIN (GenBank accession number KJ956809). The Open Reading Frame (ORF) of CfCIN is 1 161bp in length and encodes a protein with 386 amino acids. To investigate the specific functions of CfCIN, we constructed the expression vector and introduced it into Saintpaulia ionantha by Agrobacterium mediated method. Phenotypic analysis showed that the leaves of transgenic plants were bigger and changed from circular to oval. Margin of leaves changed from flat and smooth to slightly notched and back curly. Leaf veins were obvious and petioles were redder than wild type plants. There was no obvious difference in flower shape between the transgenic plants and wild type plants. These result indicated that CfCIN might participate leaf development.