雪花莲凝集素基因转化菊花及转基因植株的抗蚜性研究

针对菊花存在的蚜虫虫害问题，采用农杆菌介导法将gna基因导入菊花叶片，共获得93个转化克隆。研究了影响转化频率的主要因素，得出在使用pH5．6的YEB培养基，菌液浓度OD600=0．4，45日苗龄中部叶片预培养1d，共培养4d，共培养的培养基中加入0．5mg／L GA3的条件下可使转化频率提高到11．21％。PCR、实时荧光PCR检测结果表明，外源基因已整合到植物细胞基因组中。转化植株幼苗饲虫实验表明，不同转化克隆的抗蚜性差异较大，蚜口密度抑制率从10％—84％不等，平均蚜口密度抑制率为39．4％。转化植株叶片蛋白提取液对小鼠红细胞具有凝集作用。

Study on Transformation of Snowdrop Lectin Gene to Chrysanthemum and Aphid Resistance of the Transgenic Plants

Agrobacterium-mediated transformation in chrysanthemum was studied to prevent the insect pest of aphid (Mizus persicae).The gna gene was successfully transferred into chrysanthemum by leaf dish,and 93 transgenic clones were obtained.The highest transformation frequency 11.21% was achieved on the optimization facts,which were medium YEB with pH5.6,bacterial concentration OD_(600)=0.4,precultivation for one day ,cocultivation for four days,the cocultivation media supplemented with GA_3 0.5 mg/L and leaf explants growed for 45 days.The results from PCR and FQ-PCR analysis confirmed that gna gene was intergrated into the genome of chrysanthemum plants.The insect bioassay with aphid showed that the aphid resistance of different transgenic plants was difference,and the rate of aphid population inhibition of them were from 10% to 84% with an average rate of 39.4%.The leaf-extracts from different transgenic plants showed varying actinties in red-blood cell bioassay.