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首页> 外文期刊>Frontiers of Agriculture in China >Construction of RNAi vector for flower-related gene and verifications of the mutant in Arabidopsis thaliana
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Construction of RNAi vector for flower-related gene and verifications of the mutant in Arabidopsis thaliana

机译:花相关基因RNAi载体的构建及拟南芥突变体的验证

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摘要

The timing of floral transition is tightly controlled by a combination of endogenous and environmental signals. One early flowering mutant plant was screened from Arabidopsis library of T-DNA insertion to accelerate flowering under short-day condition, and a related-gene EFS1 (AT4G36680.1) was isolated and identified as a novel flowering-time gene of Arabidopsis in our preliminary studies. To investigate the function and the specific mechanism of EFS1 in the flower process control, the RNAi expression vector containing EFS1 gene-specific sequences in the sense and antisense orientations was constructed and transferred into Arabidopsis by using the floral-dip method, with 11 transgenic plants obtained through hygromycin B screening and PCR assays. The results showed that the expression level of EFS1 in transgenic lines was significantly lower than that in wild type and efs1 mutant. The flowering time of the efs1 mutant and RNAi transgenic plants was much earlier than that of wild-type plants. This result further verified that the EFS1 gene played an important role in flowering, and its specific mechanisms need further study. These work provided a foundation to further regulatory mechanisms of EFS1 in the control of floral transition.
机译:花转变的时机受内源性和环境信号的严格控制。从拟南芥T-DNA插入文库中筛选出一株早花突变植物,以在短时条件下加速开花,并分离了一个相关基因EFS1(AT4G36680.1),并将其鉴定为拟南芥中一个新的开花时间基因。初步研究。为了研究EFS1在花过程控制中的功能和特异性机制,构建了含有EFS1基因有义和反义方向序列的RNAi表达载体,并通过花浸法将其转入拟南芥中,并与11株转基因植物一起转移到拟南芥中。通过潮霉素B筛选和PCR分析获得。结果表明,转基因品系中EFS1的表达水平明显低于野生型和efs1突变体。 efs1突变体和RNAi转基因植物的开花时间比野生型植物的开花时间要早得多。该结果进一步证实了EFS1基因在开花中发挥了重要作用,其具体机制有待进一步研究。这些工作为进一步控制EFS1调控花序过渡提供了基础。

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