论文部分内容阅读
[目的]分析烟草中LTR类反转录转座子对植物生长发育的影响。[方法]利用同源克隆及RACE的方法,从烟草栽培品种‘贵烟1号’c DNA中克隆得到了1个烟草反转录转座子的全长c DNA序列,命名为Ntrt1(Gen Bank登录号:KC899077)。利用实时定量RT-PCR研究了该基因在烟草不同组织中的表达水平;构建了基于双生病毒卫星诱导的Ntrt1的基因沉默载体,以中国番茄黄化曲叶病毒(tomato yellow leaf curl China virus,TYLCCNV)为辅助病毒在烟草中沉默了该基因,并调查了基因沉默后的表型。[结果]序列分析表明:该基因全长5 046 bp,编码3个ORF,分别为Gag/pol多聚蛋白、反转录转座子的中心蛋白以及一个整合酶。该基因能在烟草的根、茎、叶中表达,在叶片中的表达量最高。Ntrt1基因沉默烟草与对照相比,烟草植株明显变矮。[结论]Ntrt1在烟草的生长发育中具有重要作用。
[Objective] The research aimed to analyze the effect of LTR retrotransposon in tobacco on plant growth and development. [Method] The full-length c DNA sequence of a tobacco retrotransposon was cloned from the c DNA of tobacco cultivar ’Guiyan 1’ by homologous cloning and RACE and named as Ntrt1 (Gen Bank Accession number: KC899077). The expression level of this gene in different tissues of tobacco was investigated by real-time quantitative RT-PCR. A gene silencing vector of Ntrt1 induced by the geminivirus satellite was constructed. The gene was cloned from tomato yellow leaf curl China virus (TYLCCNV) ) As a helper virus silenced the gene in tobacco and investigated the phenotype after gene silencing. [Result] Sequence analysis showed that the gene was 5 046 bp in length and encoded three ORFs, which were the Gag / pol polyprotein, the center protein of retrotransposon and an integrase. The gene can be expressed in the roots, stems and leaves of tobacco and has the highest expression in the leaves. Tobacco plants with Ntrt1 gene silencing were significantly shorter compared to the control. [Conclusion] Ntrt1 played an important role in the growth and development of tobacco.