一个参与拟南芥DNA去甲基化过程候选基因的定位与功能分析

发布时间：2018-02-02 12:08

本文关键词： 拟南芥 rll 图位克隆 DNA去甲基化　出处：《植物生理学报》2017年04期 　论文类型：期刊论文

【摘要】：以转基因拟南芥(Arabidopsis thaliana)Col-LUC为亲本材料,把它的种子进行甲基磺酸乙酯(EMS)诱变,在M2代幼苗中筛选出一株荧光降低的候选突变体,命名为rll1(reduced LUC luminescence 1),此突变基因对应的野生型基因位点因此被命名为At RLL1位点。该突变体在生长发育方面表现出明显矮化、角果较短、叶片较小的发育缺陷表型。遗传学分析表明该突变属于细胞核单基因隐性突变。通过图位克隆技术把At RLL1位点定位于2号染色体T3K9克隆。酶切PCR(chop-PCR)结果表明该突变体中35S启动子和基因组DNA上的多个位点均表现出甲基化升高的现象。反转录PCR(RT-PCR)结果表明rll1中一些RNA介导的DNA甲基化(Rd DM)途径的内源靶基因的表达量明显下降。综上所述,这些研究结果表明At RLL1位点很可能参与了拟南芥DNA去甲基化过程。
[Abstract]:The seeds of Arabidopsis thaliana)Col-LUC were mutated by ethyl methyl sulfonate (EMSs). A candidate mutant, named rll1(reduced LUC luminescence 1, was selected from M2 seedlings. The wild-type gene site corresponding to the mutant gene was named at RLL1 locus. The mutant showed obvious dwarfism in growth and development and short pods. Small developmental defect phenotype in leaves. Genetic analysis showed that the mutation was a recessive mutation of nuclear single gene. The RLL1 locus was located on chromosome T3K9 of chromosome 2. The results of chop-PCR showed that the promoter of 35s and multiple loci of genomic DNA in the mutant showed the phenomenon of increased methylation. Reverse transcriptional PCR- RT-PCR). The results showed that the expression of endogenous target genes in some RNA mediated DNA methylation (Rd) pathway in rll1 was significantly decreased. These results suggest that the at RLL1 site may be involved in the DNA demethylation process in Arabidopsis thaliana.
【作者单位】：南京农业大学生命科学学院;
【基金】：南京农业大学中央高校基本科研业务费(KYRC201409)~~
【分类号】：Q943.2
【正文快照】： 收稿2017-02-20修定2017-03-22资助南京农业大学中央高校基本科研业务费(KYRC201409)。在高等真核生物中,表观遗传修饰调控基因表达,在个体发育、病毒抗性、转座子沉默和转基因表达等方面均发挥着重要的作用(Amedeo等2000)。在高等植物中,基因沉默的类型可以分为转录水平基因

【相似文献】