农杆菌介导的无籽基因转化诺丽根段  

Transformation of non-seeded gene into noni root segment by Agrobacterium

作　　者：刘娟 蓝增全 吴田 杨自云 Juan Liu;Zengquan Lan;Tian Wu;Ziyun Yang(College of Horticulture and Gardening,Southwest Forestry University,Southwest Landscape Arrchitecture Engineering Research Center of State Forestry Administration,Kunming 650224,China;Southwest Green Development Institute,Southwest Forestry University,Kunming 650224,China)

机构地区：[1]西南林业大学园林学院,国家林业局西南风景园林工程技术研究中心,云南昆明650224 [2]西南绿色发展研究院,云南昆明650224

出　　处：《生物技术》2018年第5期473-477,466,共6页Biotechnology

基　　金：云南省应用基础研究计划项目(“ACO基因在诺丽果实成熟过程中的作用机制研究”,No.2016FB049)

摘　　要：[目的]为获得无籽诺丽植株。[方法]实验利用含有无籽基因的农杆菌对诺丽根段进行遗传转化,诺丽根段经预培养3 d、农杆菌液侵染20 min、共培养基暗培养3 d、筛选培养20 d后得到抗性芽,继代筛选后得到转化子;通过提取诺丽转化子叶片的DNA,以NptⅡ为引物进行PCR检测;将阳性植株进行生根培养后,炼苗并移栽至营养钵,观察其生长状况。[结果]表明:诺丽根段经过筛选培养后,获得225株抗性芽,抗性芽平均分化率为56. 25%;抗性芽继代筛选培养后得到94株转化子,对其进行PCR检测,获得18株转基因阳性植株;移栽后,能正常生根且于营养钵中正常生长。[结论]经PCR初步鉴定,无籽基因成功导入诺丽植株中,转化率为8%。[ Objective ] To obtain seedless noni plants. [ Methods ] An Agrobacterium containing seedless genes is used in the experiment to carry out genetic transformation on the noni root segment. Noni root segment is precuhured for 3 d,infected with Agrobacterium solution for 20 min, co - culture of dark culture for 3 d, and screened for 20 d to obtain resistant bud, and then get transformed seeds after secondary screening. By extracting DNA of blades of noni transformed seeds＇ leaves,Npt Ⅱ as primers for PCR detection; after the positive plants were taken root culture,the seedlings were refined and transplanted to the nutrition bowl to observe their growth status. [ Results ] show that after the noni root segment is screened and cultured,225 resistant buds are obtained,and the average differentiation rate of resistant buds is 56.25%. 94 transformants are obtained after the secondary culture of resistant buds, which are detected by PCR and 18 transgenic positive plants are obtained. After transplantation, it can take root and grow normally in the nutrition bowl. [ Conclusion ] the seedless gene is successfully introduced into noni plants, according to primary evaluation of PCR,and the conversion rate is 8%.

关 键 词：农杆菌侵染 诺丽 抗性芽 遗传转化 无籽基因 

分 类 号：Q812[生物学—生物工程]