大叶落地生根STM基因植物表达载体构建及烟草转化

Plant Expressing Vector Construction of Kalanchoe daigremontiana STM Gene and Its Transformation into Tobacco

为了研究大叶落地生根中(Kalanchoe daigremontiana)的KdSTM基因对侧芽的形成具有一定的促进作用。采用RT-PCR技术以大叶落地生根叶片边缘的小植株的总cDNA为模板,克隆得到KdSTM基因。KdSTM开放阅读框750 bp,编码249个氨基酸残基。为进一步研究该基因是否具有促进植物芽再生的作用,构建了p35S::KdSTM植物过表达载体,利用农杆菌介导将植物表达载体侵染烟草,转化植株经分子鉴定.成功获得转基因烟草植株。通过对转基因烟草试管苗和野生型试管苗的表型观察,发现二者的表型存在有很大的差异。转基因型烟草的叶片浓绿,早期出现较小分枝侧芽,而野生型未出现侧芽。本研究的结果表明克隆的大叶落地生根中KdSTM基因具有生物学活性,对侧芽的形成具有一定促进的作用。

To study the function of Kalanchoe daigre/nontiana KdSTM gene on the formation of lateral buds.Through KTPCR, using plantlet cDNA of Kalanchoe daigremonliana as VCR template,the experiment cloned KdSTM gene.which contains 750 bp open reading frame encoding 249 amino acid residues.To further study the stimulating function of the gene on the regeneration of buds,the over-expression vector of p35S：：KdSTM was constructed.The vector was transferred into tobacco via Agrobacterium-medmtcd transformation.Transgenic plants were thus obtained.Phenotypic observation found that transgenic tobacco plants and wild-type tobacco plants differed markedly.Transgenic tobacco plants have dark green leaves and early emerged smaller lateral buds;wild-type did not have lateral buds.The results show that KdSTM gene in the cloned Kalanchoe daigremontiana has biological activity,which plays a certain role in promoting the formation of lateral buds.