石蒜凝集素基因转化切花菊及抗蚜性鉴定  

Transformation of Lycoris Longituba Agglutinin Gene to Cut Chrysanthemum and Identification of Aphid Resistance in the Transgenic Plants

作　　者：何俊平[1] 陈发棣[1] 陈素梅[1] 房伟民[1] 缪恒彬[1] 罗火林[1] 

机构地区：[1]南京农业大学园艺学院,南京210095

出　　处：《西北植物学报》2009年第11期2318-2325,共8页Acta Botanica Boreali-Occidentalia Sinica

基　　金：江苏省科技支撑计划(BE2008307);教育部留学归国人员启动基金(2008890);中国博士后基金(20070411058);教育部新世纪优秀人才支持计划(NCET-060489);江苏省青蓝工程优秀青年骨干教师(2008[30])

摘　　要：以切花菊‘神马’叶片为外植体,采用根癌农杆菌介导法将长筒石蒜凝集素基因LLA导入菊花。结果表明,‘神马’叶片对潮霉素（Hyg）十分敏感,叶盘再生筛选以810 mg.L-1为宜,生根筛选以18 mg.L-1为宜;羧苄青霉素抑菌浓度早期以500 mg.L-1为宜,后期以250300 mg.L-1为宜。Hyg抗性再生植株经PCR、RT-PCR检测初步证实外源基因已转入7个转化株系的植物基因组DNA中并发生转录。转基因植株幼苗人工接种蚜虫实验表明,不同转化株系的抗蚜性差异较大,蚜口密度抑制率为12.2%76.8%不等,平均蚜口密度抑制率为41.8%。Using leaf disks of cut-flower chrysanthemum ＇Jinba＇ as explants,the LLA gene was successfully transferred into chrysanthemum via Agrobacterium-mediated leaf transformation.The results showed that the leaf was very sensitive to hygromycin（Hyg） and carbenicillin（Carb）,the optimal concentration of Hyg for callus selecting was 8～10 mg·L-1,the optimal selective concentration for rooting selection was 18 mg·L-1.To control agrobacterium growth,the optimal concentration of Carb was 500 mg·L-1 for early multiplication controlling,while 250～300 mg·L-1 for late control.PCR and RT-PCR analysis in Hyg-resistant lines implied that the exogenous gene LLA has been likely integrated into the genome of transformed chrysanthemum and expressed successfully.The aphid inoculation test showed that the aphid resistance of different transgenic plants was varied,and the rate of aphid population inhibition ranged from 12.2% to 76.8% with an average inhibition rate of 41.8% in transformed chrysanthemum.

关 键 词：切花菊 根癌农杆菌 长筒石蒜凝集素基因 遗传转化 蚜虫 

分 类 号：Q789[生物学—分子生物学]