柠檬苦素类似物糖基转移酶基因(citLGT)在转基因锦橙中的异位表达分析  

Ectopic expression analysis of Limonoid UDP-glucosyltransferase gene(citLGT)in transgenic Citrus sinensis ‘Jincheng'

作　　者：马园园[1,2] 邹修平[2] 彭爱红[2] 许兰珍[2] 何永睿[2] 陈善春[2] 

机构地区：[1]西南大学园艺园林学院,重庆400716 [2]西南大学柑桔研究所.国家柑桔工程技术研究中心.国家柑桔品种改良中心,重庆400712

出　　处：《果树学报》2014年第2期181-186,I0001,共7页Journal of Fruit Science

基　　金：国家科技支撑计划课题任务书(项目编号:2012BAD19B00);柑橘产业技术体系(CARS-27);农业部公益性行业(农业)科研专项(2010 03067-02-4);柑橘学重庆市市级重点实验室开放基金计划项目(CKLC201105);中央高校基本科研业务费专项(XDJK2012B023)

摘　　要：【目的】分析柠檬苦素类似物糖基转移酶基因(citLGT)在柑橘中的异位表达。【方法】构建了CaMV 35S启动子控制下citLGT的超量和RNAi抑制表达载体。采用农杆菌介导的遗传转化方法将上述植物表达载体分别转化‘锦橙’[Citrus sinensis(Linn.)Osbeck‘Jincheng’]上胚轴。经GUS组织化学染色及PCR检测共获得20株转基因阳性植株。利用qRT-PCR分析转基因植株中citLGT基因的表达。【结果】超量表达citLGT的转基因植株中citLGT的表达量显著高于对照,而RNAi转基因植株中citLGT的表达量则被显著抑制。【结论】本实验为阐明了citLGT基因影响后苦味性状的分子机制和基因工程改良柑橘延迟苦味性状提供了有价值的材料。[Objective]The goal of this study was to analyse citrus eetopie expression of Limonoid UDP- glucosyhransferase gene（citLGT） in Citrus sinensis （Linn.） Osbeck ＇Jincheng＇. [Method]Plant expression vectors with overexpressing and RNAi-suppressing eitLGT drived by CaMV 35S promoter were constructed. Genetic transformation of ＇Jincheng＇ was performed by Agrobaeterium-mediated method. 20 transgenie plants were obtained by GUS histoehemical straining and PCR analysis. The expression analysis of citLGT gene in transgenic and nontransgenic plants were performed by qRT-PCR using actin gene as a housekeeping control. [Result]The result showed that a distinct reduction and rise in the expression of eitLGT gene was detected in the RNAi-suppressed lines and overexpressing lines, respectively, when compared with the control. [Conclusion]Our study offered some transgenic lines with up-regulation and down-regulation eitLGT gene for the illuminating of the molecular mechanism and the genetic improve- ment of delayed bitterness in future.

关 键 词：柠檬苦素类似物葡萄糖基转移酶基因 RNAI 遗传转化 表达分析 转基因'锦橙’ 

分 类 号：S666.2[农业科学—果树学]