Li Xiangqian, Gasic Ksenjia, Cammue Bruno, Broekaert Willem, Korban Schuyler S
Department of Natural Resources and Environmental Sciences, 310 ERML, University of Illinois, 1201 W. Gregory, Urbana, IL, USA.
Planta. 2003 Dec;218(2):226-32. doi: 10.1007/s00425-003-1093-5. Epub 2003 Sep 24.
An antimicrobial protein gene, Ace-AMP1, was introduced into Rosa hybrida cv. Carefree Beauty via Agrobacterium-mediated transformation. A total of 500 putative transgenic plants were obtained from 100 primary embryogenic calli co-cultivated with A. tumefaciens following selection on a regeneration medium containing 100 mg/l kanamycin. Polymerase chain reaction analysis of these putative transgenic lines, using primers for both Ace-AMP1 and neomycin phosphotransferase ( npt II) genes, showed that 62% of these plants were positive for both transgenes. These lines were further confirmed for stable integration of Ace-AMP1 and npt II genes by Southern blotting. Transcription of the Ace-AMP1 transgene in various transgenic rose lines was determined using Northern blotting. Transgenic rose lines inoculated with conidial spores of Sphaerotheca pannosa (Wallr.: Fr.) Lev. var. rosae showed enhanced resistance to powdery mildew using both a detached-leaf assay and an in vivo greenhouse whole-plant assay.
通过农杆菌介导的转化,将一种抗菌蛋白基因Ace - AMP1导入杂交茶香月季品种“无忧无虑美”。在含有100 mg/l卡那霉素的再生培养基上筛选后,从与根癌农杆菌共培养的100个初生胚性愈伤组织中总共获得了500株推定的转基因植株。使用针对Ace - AMP1和新霉素磷酸转移酶(npt II)基因的引物对这些推定的转基因株系进行聚合酶链反应分析,结果表明62%的植株这两个转基因均为阳性。通过Southern杂交进一步证实了这些株系中Ace - AMP1和npt II基因的稳定整合。使用Northern杂交确定Ace - AMP1转基因在各种转基因玫瑰株系中的转录情况。使用离体叶片试验和温室活体全株试验,接种蔷薇单囊壳菌(Wallr.: Fr.)Lev. var. rosae分生孢子的转基因玫瑰株系对白粉病表现出增强的抗性。