发根农杆菌T-DNA诱变怀地黄突变体或品系的遗传学分析  

Genetic Analyses of Rehmannia Glutinosa f.Hueichingensis Hsiao Mutants Derived from Agrobacterium RhizogenesT-DNA Transformation

作　　者：范红军[1] 姚换灵[1] 张永华[1] 周延清[1,2,3] 张喻[1] 李静云[1] 陈娟娟[1] 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]河南师范大学资源微生物与功能分子河南省高校重点实验室培育基地,河南新乡453007 [3]河南师范大学道地中药材保育及利用河南省高校工程技术研究中心,河南新乡453007

出　　处：《河南师范大学学报（自然科学版）》2013年第5期115-119,共5页Journal of Henan Normal University(Natural Science Edition)

基　　金：河南省基础与前沿技术研究计划项目(092300410009);河南省教育厅自然科学研究计划项目(科技攻关)(2011A180022);河南省教育厅自然科学研究计划项目(2011A180016);河南省高校科技创新团队支持计划资助(13IRTSTHN009)

摘　　要：主要采用紫外吸收光谱法、高效液相色谱法、3,5-二硝基水杨酸比色法,PCR和SRAP技术,从叶绿素含量、还原糖和总糖含量、梓醇含量、毛蕊花糖苷含量和T-DNA片段的稳定性和基因组DNA的变化方面,对发根农杆菌转化怀地黄突变体或品系的变异性状进行了分析.发现怀地黄突变体或品系的叶片形态、植高、叶绿素含量、糖含量、梓醇和毛蕊花糖苷的含量均有变异,一个SRAP引物组合从其基因组DNA中扩增出一个对照中没有的约400bp的DNA片段,T1-T7代突变体或品系中均检测到rolB基因的存在.这些结果表明发根农杆菌T-DNA可以诱导怀地黄产生变异,并且稳定遗传.Rehmannia glutinosa f.hueichingensis Hsiao mutants derived fromAgrobacterium rhizogenes T-DNA transformation were characterized by means of their morphology,chlorophyll content,reducing sugar and total sugar content,catalpol and verbascoside contents,Inserted T-DNA stability and DNA variation analysis using ultraviolet absorption spectrum,HPLC,3,5-Dinitrosalicylic acid colorimetry,PCR and SRAP.There were different changes in leaf morphology,plant height,chlorophyll content,total sugar content,reducing sugar content,catalpol and verbascoside contents and genomic DNA of T-DNA insertional mutants or lines from 85-5（CK）.Moreover,rolBgene existed in one of their genomic DNAs from T1to T7 generation.The results indicated that Agrobacterium rhizogenes T-DNA could cause Rehmannia glutinosa f.hueichingensis Hsiao to mutate,and was stably inherited from generation to generation.

关 键 词：怀地黄 发根农杆菌 突变体 高效液相色谱 SRAP 

分 类 号：Q788[生物学—分子生物学]