黄瓜花叶病毒外壳蛋白基因转化辣椒的研究

Transformation of Capsicum annuum with Coat Protein Gene of Cucumber Mosaic Virus

以辣椒品种‘B162’为试材,探讨了通过根癌农杆菌介导法将黄瓜花叶病毒外壳蛋白(CMV-CP)基因转化辣椒的适宜条件.结果表明,将辣椒带柄子叶进行预培养2 d后,用D600 nm为0.3的根癌农杆菌菌液浸泡7 min,再经过共培养2 d后转移到筛选分化培养基中进行培养,有利于获得辣椒抗性不定芽.通过对获得的10株抗性不定芽进行PCR检测,其中有2株扩增出目的条带,初步证明CMV-CP基因已经转入辣椒不定芽中.

Agrobacterium-mediated transformation of pepper ＇ B162＇ with coat protein gene of cucumber mosaic virus （CMV-CP） was studied. Results showed that the best transformation system was ： cotyledons explants were precultured for 2 d, and then inoculated with Agrobacterium tumefaciens （D600 nm = 0. 3 ） for 7 min, transferred onto selection differentiation medium after co-cultured for 2 d. 10 green adventitious buds with Kan-resistance were screened by Polymerase chain reaction. Two of them were amplified with the band of the fragment of 740 bp. The result showed the CMV-CP gene had been incorporated into the plant genome successfully.