黄河蜜甜瓜CMV CP基因转化及其抗病性鉴定

Plant regeneration of transgenic Huanghemi (Cucumis melo) and identification of its resistance to virus disease

利用整合有CMVCP基因及NPT-II基因的改建质粒pBim438,以农杆菌为载体,对黄河蜜甜瓜子叶进行转化,以75mg/L卡那霉素筛选转化体,获得了完整的转基因植株。Southern杂交证明转化植株整合了CMVCP基因,Western杂交证明转基因获得了表达。温室CMV攻毒试验及病毒含量测定表明,转基因甜瓜对CMV的侵染表达了较高的抗病性,能够推迟系统症状显症发生,减轻病害发生程度,转基因植株体内病毒含量低于对照。转基因植株抗病性存在差异,筛选出的2个抗性株系中,TM0-1抗性高于TM0-2。

Segments of the cotyledon from a muskmelon cultivar Huanghemi 3 was infected by Agrobacteriurn tumefaciens which carrying the disarmed Ti plasmid pBim438 integrated CMV coat protein （CMV CP） gene. The transformed calli was selected in the medium with 75 mg/L kanamycin（kan）. Southern blotting analysis showed that the transgenic plants with CMV CP gene were obtained. Western blotting analysis revealed that the target protein was expressed in the transgenic plants. The results of the resistance test showed that the transgenic plants showed certain resistance to the relevant virus, it can delay the disease infection, and reduce the percentage of showing symptom, incidence index, as well as the concentration of virus. Moreover, two transgenic plants with high virus resistance were selected, in which one transgenic plant TM0-1 showed higher virus resistance than the other transgenic plant TM0-2.