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Abstract：
Plant flowering time is affected by endogenous and exogenous factors, but its variation patterns among different populations of a species has not been fully established. In this study, 27 Arabidopsis thaliana accessions were used to investigate the relationship between autonomous pathway gene methylation, gene expression and flowering time variation. DNA methylation analysis, RT-qPCR and transgenic verification showed that variation in the flowering time among the Arabidopsis populations ranged from 19 to 55 days and was significantly correlated with methylation of the coding regions of six upstream genes in the autonomous pathway, FLOWERING LOCUS VE (FVE), FLOWERING LOCUS Y (FY), FLOWERING LOCUS D (FLD), PEPPER (PEP), HISTONE DEACETYLASE 5 (HAD5) and Pre-mRNA Processing Protein 39-1 (PRP39-1), as well as their relative expression levels. The expression of FVE and FVE(CS) was modified separately through degenerate codon substitution of cytosine and led to earlier flowering of transgenic plants by 8 days and 25 days, respectively. An accurate determination of methylated sites in FVE and FVE(CS) among those transgenic plants and the recipient Col-0 verified the close relationship between the number of methylation sites, expression and flowering time. Our findings suggest that the methylation variation of these six key upstream transcription factors was associated with the gene expression level of the autonomous pathway and flowering time in Arabidopsis. The FVE(CS) and FVE genes in transgenic plants tended to be hypermethylated, which could be a protective mechanism for plants. However, modification of gene sequences through degenerate codon substitution to reduce cytosine can avoid hypermethylated transferred genes in transgenic plants. It may be possible to partially regulate the flowering of plants by modified trans-epigenetic technology.
摘要：
植物开花时间受内源和外源因素的影响，但是它在一个物种不同种群之间的变异模式尚未完全确定。在这项研究中,27个拟南芥种质用于研究自主途径基因甲基化之间的关系，基因表达和开花时间变异。DNA甲基化分析,RT-qPCR和转基因验证表明，拟南芥种群的开花时间变化范围为19至55天，并且与自主途径中六个上游基因编码区的甲基化显着相关。花落VE(FVE),花球Y(FY),花斑D(FLD),PEPPER(PEP),组蛋白脱乙酰酶5(HAD5)和mRNA前加工蛋白39-1(PRP39-1),以及它们的相对表达水平。通过胞嘧啶的简并密码子替换分别修饰FVE和FVE（CS）的表达，并导致转基因植物在8天和25天之前提前开花。分别。这些转基因植物和受体Col-0中FVE和FVE（CS）中甲基化位点的准确测定验证了甲基化位点数量之间的密切关系，表达和开花时间。我们的发现表明，这六个关键上游转录因子的甲基化变异与拟南芥自主途径的基因表达水平和开花时间有关。转基因植物中的FVE(CS)和FVE基因趋于高甲基化，这可能是植物的保护机制。然而,通过简并密码子置换以减少胞嘧啶来修饰基因序列可以避免转基因植物中过度甲基化的转移基因。通过改良的反式表观遗传技术可能可以部分调节植物的开花。
