荠菜CBF冷诱导途径多价载体的构建和烟草转化

Construction of Cold Induction Pathway Multivalent Vector in Capsella bursa-pastoris CBF and Transformation of Tobacco

为了研究荠菜CBF抗冷途径多基因共表达对转基因植株耐寒性的影响,通过同尾酶法将CbCBF、CbICE53、CbRCI35以及CbCOR15bP进行结合,构建了CbCOR15bP::CbICE53+pCaMV35S::CbCBF+pCaMV35S::CbRCI35多价植物表达载体并进行了烟草的转化,通过形态观察以及电解质渗透率和相对水含量这两个生理指标的测定显示,转基因植株在4℃和-4℃低温处理情况下,多基因共转化的转基因植株在电解质渗透率和相对水含量与野生型和空载对照之间的差异非常明显,转基因植株在低温下的抗寒冻能力也明显增强,说明CBF途径多基因共转化可以明显提高烟草的抗寒冻能力,此项研究具有一定的实际应用意义。

To investigate the effects of multiple gene expression of CBF cold response pathway in Capsella bursa-pastoris on the cold tolerance of co-transgenic plants, we combined CbCOR15bP, CbICE53, CbCBF and CbRCI35 into a reconstruct vector by isocaudomers. After transferring the pCbCOR15bP..CbICE53-f- p35S： ：CbCBFq-p35S：：CbRCI35 vector into tobacco, we observed the morphologies of the transgenic plants and analyzed the electrolyte leakage and relative water content. The results show that two physiological indexes indicated cold-tolerance. In transgenic plants, the electrolyte leakage and relative water content were significantly different with the wild type when the plants were treated at the lower tempreture 4℃ and -4 ℃. The empty vector transgenic plants, after cold and freezing treatments,showed a much greater cold tolerance. Constructing co-transformation vectors by isocaudomers can be an effective and efficient method for genetic breeding and crop agriculture.