摘要
以哥伦比亚(Columbia)野生型拟南芥(Arabidopsis thaliana)为实验材料,用含有激活标记双元质粒pCB260的农杆菌浸花进行转化,构建拟南芥T-DNA插入突变体库。通过突变体的筛选和表型分析,获得了两株光形态突变体,子叶下胚轴伸长的光抑制效应减弱。通过TAIL-PCR(thermal asymmetric interlaced-PCR)技术,成功扩增出突变植株T-DNA插入位点侧翼序列,经NCBI序列比对,T-DNA分别插在CRY1第一和第三外显子部位。突变体的表型分析及PCR鉴定结果表明,T-DNA插入CRY1并影响到突变植株的光形态建成。
Columbia wild type Arabidopsis thaliana was used as experimental material, an activation tagging mutant library was constructed by floral dip method, Agrobacterium tumefaciens with an activation tagging vector pCB260 was transformed into the plant. By mutant screening and phenotypic analysis, two mutants were isolated. The hypocotyl elongation is weak inhibited in the mutant seedlings by continuous white lights and the T-DNA flanking sequences were obtained by TAIL-PCR. The NCBI sequence alignment indicates that T-DNA inserted in the first and third exon positions of Cryl, respectively. Mutant phenotype analysis and PCR identification results show that T-DNA insertion disrupt the function of CRY1 and abolish the process of plant photomorphogenesis.
出处
《植物分类与资源学报》
CAS
CSCD
北大核心
2011年第6期645-652,共8页
Plant Diversity
基金
国家自然科学基金面上项目(30971452)
中国科学院百人计划项目
