美洲商陆抗病毒蛋白基因遗传转化甘蔗的研究

Genetic Transformation of Pokeweed Antiviral Protein Gene into Sugarcane

构建叶片特异表达启动子rbcs驱动的美洲商陆抗病毒蛋白基因PAP-c的植物表达载体PNRPL,冻融法转化农杆菌EHA105后,通过农杆菌介导法侵染甘蔗优良品种ROC22的胚性愈伤组织,经PPT抗性筛选以及PCR鉴定,共获得了24株转化植株,对其中的5株进行Southern杂交检测,有2株呈阳性,初步证明外源基因已整合到甘蔗基因组中;对其接种甘蔗花叶病毒,初步结果表明获得的转化植株对甘蔗花叶病毒具有一定的抗性。

Pokeweed antiviral protein gene PAP-c＇s plant expression vector PNRPL was constructed, which was driven by leaf specific expression promoter rbes. The Agrobacterium tumerfaciens EHA105 was transformed by the ＂freeze and thaw＂ method, and the embryonic calli of elite sugarcane variety ROC22 were inoculated by using the A. tumefaciens-mediated method. PPT resistance screening and PCR identification results showed that 24 transformed plants were generated. Of the transformed plants 5 were taken for Southern blot analysis, 2 of which were positive, preliminarily proving that exogenous gene had been integrated into sugarcanes genome. Inoculation of sugarcane mosaic virus further proved that these transgenic plants had certain resistance on the sugarcane mosaic virus.