Establishment of the Agrobacterium-mediated Genetic Transformation System of Ginkgo biloba and the Construction of the Expression Vector of Gb-DXR  

农杆菌介导银杏遗传转化体系的建立及Gb-DXR基因表达载体的构建(摘要)(英文)

作　　者：冯国庆[1] 杨颖舫[1] 李郑娜[1] 成瑜[1] 杨春贤[1] 陈敏[1] 廖志华[1] 

机构地区：[1]西南大学生命科学学院,重庆400715

出　　处：《Agricultural Science & Technology》2010年第3期28-32,114,共6页农业科学与技术（英文版）

摘　　要：[Objective] The research aimed to provide reference for increasing the genetic transformation efficiency of Ginkgo biloba mediated by Agrobacterium.[Method] Taking the mature embryos of Ginkgo biloba seeds as explants,after 48 hours＇ pre-cultivation on MS medium in the absence of phytohormone,GUS gene was transmitted into embryos of Ginkgo biloba mediated by three kinds of Agrobacterium.Transient expression of GUS gene activity was observed through histochemical staining,and the influencing factors of the expression of GUS gene were analyzed.And the expression vector of 1-deoxy-D-xylulose-5-phosphate reductoisomerase in the biosynthesis approach of biobalide precursor of Ginkgo biloba was constructed.[Result] A more suitable genetic transformation scheme was obtained as follows：taking embryos of Ginkgo biloba as explants,using EHA105 Agrobacterium with pCAMBIA1304＋ for infection,co-culture for 3 days and GUS staining.The results showed that transient expression rate of GUS after transformation was higher.[Conclusion] The research provide a more effective method for further study on the transgene of Ginkgo biloba.[目的]为提高农杆菌介导的银杏遗传转化效率提供参考。[方法]以成熟的银杏种子胚为外植体,在无激素MS培养基上预培养48 h后,采用3种农杆菌(C58C1、LBA4404和EHA105)介导将GUS基因导入银杏胚中,经过组织化学染色法检测到GUS瞬时表达活性,对影响GUS基因表达的因素进行初步分析;并构建了银杏内酯前体合成途径上1-脱氧-D-木酮糖-5-磷酸还原异构酶(DXR)的表达载体。[结果]该研究得到较合适的遗传转化方案,即用银杏胚作为外植体,用携带pCAMBIA1304+的EHA105农杆菌进行侵染,共培养3 d,进行GUS染色,结果显示转化后GUS阳性率较高。[结论]该研究为进一步的银杏转基因研究工作奠定了基础。

关 键 词：Embryos of Ginkgo biloba AGROBACTERIUM-MEDIATED Genetic transformation GUS gene 1-deoxy-D-xylulose-5-phosphate reductoisomerase Expression vector 

分 类 号：S792.95[农业科学—林木遗传育种]