根癌农杆菌介导的花魔芋遗传转化体系的研究  

Genetic Transformation System of Amorphophallus konjac Mediated by Agrobacterium tumfaciens

作　　者：周盈[1] 林拥军[2] 柴鑫莉 孙明[1] 

机构地区：[1]华中农业大学农业微生物学国家重点实验室,武汉430070 [2]华中农业大学作物遗传改良国家重点实验室,武汉430070

出　　处：《分子植物育种》2006年第4期559-564,共6页Molecular Plant Breeding

基　　金：国家973课题(2003CB114201);国家自然科学基金(30270053);国家863(2004AA214092);教育部重大项目资助.

摘　　要：本研究以潮霉素抗性基因为选择标记,建立了根癌农杆菌介导的花魔芋(Amorphophalluskonjac)的遗传转化体系。同时探讨了魔芋愈伤组织的培养基、筛选剂和筛选时间等因素对转化效率的影响。通过筛选时间不同的两个转化试验,分别从300个花魔芋愈伤组织中得到了16个和18个具有潮霉素抗性的愈伤组织,其抗性愈伤率为5.33%和6.00%。获得的抗性愈伤组织进一步在分化培养基上进行分化,其分化效率分别为31.2%和22.2%。移栽后,分别得到了39株和36株成活植株。PCR检测表明分别有26株和21株再生花魔芋出现了800bp的特异性扩增带型,其阳性率分别为66.7%和58.3%。随机挑取20个PCR阳性植株的DNA进行斑点杂交检测,结果表明外源基因已经整合到花魔芋的基因组中。An Agrobacterium tumfaciens-mediated transformation system has been developed for the production of transgenic plants of A morphophallus konjac. The explants were inoculated with an Agrobacterium tumfaciens strain EHA105 carrying the plasmid pBMB0890 containing selective gene hygromycin phosphotransferase （hpt）. Factors that influence Agrobacterium tumfaciens-mediated transformation of A morphophallus konjac, including culture medium （Table 1）, selection agents and selection time were investigated. 16 and 18 resistant calli were obtained from 300 calli separately. The transformation frequency was 5.33% and 6.00%, repectively. 31.2% and 22.2% of resistant calli could generate plants. PCR analysis of the 39 and 36 live plants indicated 26 and 21 plants were integrated with foreign DNA separately, resulting 66.7% and 58.3% transgenic ratio, respectively. 20 plants of those transgenic plants were re-clarified by spot blot hybridization.

关 键 词：花魔芋 根癌农杆菌 遗传转化体系 

分 类 号：S632.9[农业科学—蔬菜学]