根癌农杆菌介导北海道黄杨遗传转化体系的建立  

Establishment of Genetic Transformation System of Euonymus japonicus 'Cu Zhi' Mediated by Agrobacterium tumefaciens

作　　者：尚爱芹[1] 田传卫[1] 赵梁军[1] 田颖川[2] 

机构地区：[1]中国农业大学观赏园艺与园林系 [2]中国科学院微生物研究所,北京100080

出　　处：《园艺学报》2008年第3期409-414,共6页Acta Horticulturae Sinica

基　　金：国家博士点基金项目(20060019043);北京市科技计划项目(H020620110130)

摘　　要：在建立北海道黄杨(Euonymus japonicus ‘CuZhi’)再生体系的基础上,建立其遗传转化体系,以获得转雪花莲外源凝集素(Galanthus nivalis agglutinin,GNA)基因植株。采用根癌农杆菌(Agr-robacterium tumefaciens,LBA4404菌株)介导法,研究影响北海道黄杨遗传转化的若干因素。结果表明,预培养与否、菌液浓度、侵染时间、共培养时间等对转化频率都有一定的影响。在遗传转化过程中,不经预培养,根癌农杆菌浓度OD600=0.6,侵染下胚轴40min,再共培养3d,有利于获得最高遗传转化效率。抗性植株经PCR和DNA-Southern blot检测表明,目的基因已整合到北海道黄杨基因组中。In order to provide genetically improved new germplasm resources and achieve the GNA transgenie plants of Euonymus japonicus ＇ Cu Zhi＇ , genetic transformation system was established based on the system of regeneration. Several factors affected genetic transformation of Euonymus japonicus ＇ Cu Zhi＇ mediated by Agrobacterium tumefaciens （LBA4404） were studied. The results showed that preculture or not, bacterial concentrations, the time of infection and coultivation had some effect on transformation frequency. The highest transformation frequency was obtained through the following transformation procedure after preculture for 0 d, the explants were infected for 40 min with the concentration of Agrobacterium tumefaciens OD600 = 0. 6, and then co-cultivated for 3 d. The PCR and PCR-Southern blotting analysis showed the snowdrop lection gene was integrated into the genome of Euonymus japonicus ＇ Cu Zhi＇ The system of genetic transformation of Euonymus japonicus ＇ Cu Zhi＇ ＇ could provide new germplasm resource used in genetic improvement of Euonymus japonicus ＇ Cu Zhi＇ and the other plants of Euonyrnus.

关 键 词：北海道黄杨 雪花莲外源凝集素基因 遗传转化 

分 类 号：S687[农业科学—观赏园艺]