rolC基因对转Ri质粒三倍体毛白杨的遗传转化  

Genetic Transformation of Ri Plasmid Transgenic Plant of the Triploid of Populus tomentosa with rolC Gene

作　　者：苏彦苹[1] 刘兴菊[1] 周怀军[1] 杨敏生[1] 梁海永[1] 

机构地区：[1]河北农业大学林学院,河北保定071001

出　　处：《西北林学院学报》2007年第5期57-61,共5页Journal of Northwest Forestry University

基　　金：河北省自然科学基金(C2006000464)

摘　　要：通过对Ri质粒转化三倍体毛白杨植株转入rolC基因,使外源基因rolC在植物体内形成多拷贝,从而对rolC基因表达产生干扰,而减少Ri质粒转化三倍体毛白杨造成的不良形态变异。本研究建立了转Ri质粒三倍体毛白杨的叶片再生体系,筛选出叶片再生的最佳培养基为:MS+6-BA 1.0 mg.L-1+NAA 0.05 mg.L-1;确定了卡那霉素(Km)和抑菌抗生素头孢噻肟钠(CTX)的临界浓度为30 mg.L-1和400 mg.L-1;采用农杆菌介导的叶盘法,将rolC基因转入经发根农杆菌Ri质粒30148转化的三倍体毛白杨植株中,获得了18个Km抗性系,对其中14个株系进行了PCR检测,表明rolC基因已整合进再生植株中。与转Ri质粒三倍体毛白杨和未转基因三倍体毛白杨相比,部分转Ri质粒三倍体毛白杨在转rolC基因后发生了形态特征变异,比如部分无性系的叶片由原来的皱缩变为平展,部分无性系的茎由原来的直立变为弯曲,侧枝增多,节间距明显缩短。In this study,the rolC gene cloned from Agrobacterium rhizogenes was inserted into the genome of Ri plasmid transformation plant of the triploid of P.tomentosa by Agrobacteriunm-mediated leaf disc transformation method for decreasing the unfavourable phenotypic variation of Ri plasmid transformation plant of the triploid of P.tomentosa.Regeneration system for Ri plasmid transformation plant of the triploid of P.tomentosa leaves was established and the results showed that the optimal medium compositions for adventitious buds regeneration were MS＋6-BA 1.0 mg·L-1＋NAA 0.05 mg·L-1.Kanamycin and Cefotaxime sodium（CTX） sensitive tests showed that the critical sensitive concentration for inducing shoots was 30 mg·L-1 and 400 mg·L-1,respectively.Eighteen transgenic plantlets with kanamycin resistance were obtained and fourteen were verified by PCR amplification.Insertion of rolC gene resulted in phenotypic variation of part of transgenic plants.Part of transgenic clones showed horizontal leaves from wrinkle leaves,zigzag stem from erect stem,increase of side shoots in the stem and shortening stem internodes compared with Ri plasmid transgenic plant and untransgenic plant.

关 键 词：ROLC基因 转化 形态变异 

分 类 号：S792.117.01[农业科学—林木遗传育种]