基于GUS基因瞬时表达优化云南杜鹃(Rhododendron yunnanense Franch.)遗传转化方法  

Optimize the Genetic Transformation Method of Rhododendron yunnanense Franch. with Transient Expression of GUS Gene

作　　者：彭绿春[1,2,3] 周微 汪玲敏 张露[1,2,3] 宋杰[1,2,3] 解玮佳[1,2,3] 关文灵[5] 李世峰[1,2,3] 

机构地区：[1]云南省农业科学院花卉研究所,云南昆明650205 [2]云南省花卉育种重点实验室,云南昆明650205 [3]云南花卉技术工程研究中心,云南昆明650205 [4]九江农业科学院,江西九江332101 [5]云南农业大学园林园艺学院,云南昆明650201

出　　处：《云南农业大学学报（自然科学版）》2016年第6期1045-1051,共7页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家观赏园艺工程技术研究中心项目(2012FU125X10);云南省科技计划项目(2015BB013);云南省技术创新人才培养配套项目(2014-HB057);云南省花卉育种重点实验室开放基金(FKL-201202)

摘　　要：本研究以云南杜鹃无菌叶片和愈伤组织为受体材料,运用正交试验设计,通过组织染色分析法研究了预培养时间、菌液密度、侵染时间、共培养时间对农杆菌介导的GUS基因瞬时表达的影响。结果表明:叶片是云南杜鹃较理想的遗传转化受体材料,无菌叶片预培养0 d,菌液OD_(600)值0.5,侵染时间40 min,共培养5 d,GUS基因瞬时表达率最高,平均表达率为70%。试验结果为云南杜鹃遗传转化研究提供了快速可靠的依据。The experiment was conducted to investigate the transient expression mediated by Agrobacte- rium tumefaciens using sterile leaves and calli of Rhododendron yunnanense as receptor materials. Effects of pre-cuhure length, A. tumefaciens concentration, infection time and co-culture length on transient expression of GUS gene were studied by tissue staining method. The results showed that leaf was the optimal receptor material for genetic transformation in R. yunnanense, while the highest transi- ent expression rate （70%） of GUS gene was obtained as pre-cuhure for 0 day, OD600 value of A. tu- mefaciens for infection 0. 5, infection for 40 min, and co-culture for 5 days. The results provided a fast and reliable method for genetic transformation of R. yunnanense.

关 键 词：云南杜鹃 GUS基因 瞬时表达 遗传转化方法 

分 类 号：S685.210.36[农业科学—观赏园艺]