WANG Jing,TANG Jing,CUI Yueting,et al.Establishment of Agrobacterium-mediated Genetic Transformation System in Lycium ruthenicium Murr.[J].Northern Horticulture,2020,44(01):104-110.[doi:10.11937/bfyy.20191730]
农杆菌介导的黑果枸杞遗传转化体系的建立
- Title:
- Establishment of Agrobacterium-mediated Genetic Transformation System in Lycium ruthenicium Murr.
- 文献标志码:
- A
- 摘要:
- 以黑果枸杞下胚轴为试材，采用农杆菌浸泡侵染法，建立简单有效的黑果枸杞遗传转化体系，以期为深入研究黑果枸杞基因功能提供参考依据。结果表明：成功构建pORE R1-[STBX]35S:GUS:NOS[STBZ]质粒载体，并通过冻融法导入农杆菌GV3101。调整菌液浓度至OD600=0.5，侵染黑果枸杞下胚轴，经选择培养基筛选、PCR及GUS检测后，获得转基因阳性植株，转化阳性率为16%，通过驯化移栽后转基因植株均可成活。表明可通过农杆菌GV3101侵染黑果枸杞下胚轴实现外源基因的转化，且转化率较高。
- Abstract:
- Using Lycium ruthenicium Murr.hypcotyls as material,a method of Agorobacterium-mediated genetic transformation system for further analyzing gene function was established，in order to provide a reference for further study on the function of Lycium ruthenicium gene.The results showed that the vector pORE R1-[STBX]35S:GUS:NOS[STBZ] was constructed and transformed into Agorobacterium GV3101.Hypocotyls of Lycium ruthencum Murr.were infected with Agrobacterium suspension,OD600=0.5.After selective cultivation,PCR and GUS activity detection,identified the positive plants,with transformation rate was 16%.Transformed plants were all well growing.The results indicated that Agrobacterium could successful transform the genes into hypocotyls of Lycium ruthenicium Murr.,with highly transformation rate.
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备注/Memo
第一作者简介：王静（1984-），女，博士，讲师，现主要从事植物逆境生物化学与分子生物学等研究工作。E-mail：wangjing_imu@163.com.基金项目：宁夏自然科学基金资助项目（2018AAC3248）；北方民族大学重点科研资助项目（2019KJ19）。收稿日期：2019-09-04