雪花莲凝集素基因转化菊花及转基因植株的抗蚜性研究  

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

作　　者：王关林[1] 刘彦泓 郭绍华[1] 王宇[1] 纪彦[1] 方宏筠[1] 

机构地区：[1]辽宁师范大学生命科学院,大连116029 [2]大连市产品质量监督检验所,大连116021

出　　处：《Acta Genetica Sinica》2004年第12期1434-1438,共5页

基　　金：国家自然科学基金项目 (编号 :3 0 170 488)~~

摘　　要：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OD60 0 =0 4 ,4 5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/LGA3 的条件下可使转化频率提高到 11 2 1%。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 %。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.

关 键 词：菊花 雪花莲凝集素基因 根癌农杆菌 转化 蚜虫 

分 类 号：Q943.2[生物学—植物学]