摘要
芍药是乙烯敏感型花卉,乙烯受体感知并传导乙烯信号,在乙烯信号转导途径中发挥重要作用。芍药PlETR1基因cDNA全长序列已分离,为了鉴定芍药PlETR1基因的功能,本研究基于PlETR1基因和表达载体序列,应用Primer Premier 5.0软件设计了一对特异性PCR引物,采用RT-PCR技术扩增出了PlETR1编码区片段,进一步构建了芍药PlETR1基因过表达载体。基于优化的模式植物烟草的组培体系和筛选出的潮霉素抗性浓度,应用农杆菌介导的叶盘法开展了芍药PlETR1基因转化烟草的研究,对转基因抗性烟草植株进行了PCR检测,结果表明HPT基因和芍药PlETR1基因已导入到烟草基因组中,且芍药PlETR1基因转录表达成功,为下一步鉴定芍药PlETR1基因的功能提供科学依据。
Herbaceous peony(Paeonia lactiflora) is one of the ornamental plants which are sensitive to ethylene.Ethylene signal is percepted and transmitted by ethylene receptor,which plays an important role in ethylene signal transduction pathway.The full-length cDNA sequence of PlETR1 has been isolated from Paeonia lactiflora.In order to characterize the function of PlETR1,a specific pair of PCR primers were designed with Primer Premier5.0 software,which was based on the sequences of PlETR1 and the expression vector pCambia 1301-UbiN.The coding sequence of PlETR1 was successfully amplified with RT-PCR technique,and PlETR1 overexpression vector was constructed then.The transformation of PlETR1 into tobacco was carried out by Agrobacterium mediated leaf disc method,which was based on the optimized tissue culture system of tobacco and the screened hygromycin resistance concentration.The PCR results of the transgenic resistant tobacco plants showed that HPT and PlETR1 had been introduced into tobacco genome,and PlETR1 was successfully expressed at the transcription level,which lays a foundation for further identification of the function of PlETR1.
作者
高水平
徐杰
张文婷
吴建梅
李芳
史国安
范丙友
Gao Shuiping;Xu Jie;Zhang Wenting;Wu Jianmei;Li Fang;Shi Guoan;Fan Bingyou(College of Forestry,Henan University of Science and Technology,Luoyang,471000;College of Agriculture,Henan University of Science and Technology,Luoyang,471000;College ofBiological Sciences and Technology,Beijing Forestry University,Beijing,100083;Luoyang Wangcheng Park,Luoyang,471003;Luoyang Academy of Agriculture and Forestry,Luoyang,471023)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2020年第1期207-215,共9页
Genomics and Applied Biology
基金
国家自然科学基金(U1204323)
河南省科技攻关项目(172102110238)
河南省高等学校重点科研项目(14A220004和20A210010)共同资助。
关键词
芍药
PlETR1
过表达载体
烟草
转化
Paeonia lactiflora
PlETR1
Overexpression vector
Tobacco
Transformation
