长期培养的黄瓜毛状根中外源基因遗传稳定性分析  

Genetic Stability Analysis of Exogenous Gene in Long-term Cultured Cucumber Hairy Roots

作　　者：曹庆丰[1] 向太和[1] 孟莎莎[1] 王沙沙[1] 陆文怡[1] 

机构地区：[1]杭州师范大学生命与环境科学学院,杭州310036

出　　处：《园艺学报》2012年第8期1583-1588,共6页Acta Horticulturae Sinica

基　　金：浙江省自然科学基金项目(Y3080184);杭州市科技发展计划项目(20110232B10)

摘　　要：对在固体培养基上培养3年的黄瓜转gfp基因毛状根进行gfp和rol位点系列基因的PCR扩增、gfp的荧光定量PCR、Western blot杂交以及荧光观察分析。结果显示,由组成型启动子35S驱动的gfp在转录水平上能正常表达,而且能够翻译出编码蛋白;此外,培养3年后的毛状根,能扩增出与毛状根形态构成有关的rol系列基因。本研究结果表明长期培养的毛状根能保持其遗传稳定性,这为利用毛状根长期工厂化生产外源基因表达的蛋白产物和药用植物的活性成分提供了理论依据。In this study, it was investigated that the genetic stability ofgfp gene in the cucumber hairy roots that had been cultured on solid medium for 3 years. The analysis of PCR for gfp and rol （root loci） genes, qRT-PCR for gfp gene and Western blot for GFP protein were conducted, while green fluorescence was checked on hairy roots. The results showed that the expression of gfp driven by constitutive promoter 35S was relatively stable at both mRNA and protein levels. Furthermore, the morphology-related rol genes of the hairy roots after 3-year culturing can be amplified. Overall, this study provided important experimental supports for using hairy roots in the long-term industrial production of proteins and active ingredients of medicinal plants.

关 键 词：黄瓜 毛状根 基因 表达 稳定性 

分 类 号：S642.2[农业科学—蔬菜学]