ECT6和ECT7调控拟南芥开花时间的研究
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中国博士后科学基金 (2019M662183)


Study on ECT6 and ECT7 Involving Flowering Time of Arabidopsis
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    摘要:

    m6A修饰是mRNA上含量最丰富的一种修饰,调控mRNA的命运决定。YT521B同源性(YTH)结构域蛋白是一类典型的m6A“阅读器”,能识别并结合m6A,完成基因的转录后调控。为了探究拟南芥YTH结构域蛋白ECT6和ECT7的功能及其分子机制,该研究对ECT6、ECT7和ECT6 ECT7双突变体进行基因型和半定量PCR鉴定及开花表型观察,通过细胞学观察分析ECT6和ECT7的亚细胞定位,并采用qRTPCR检测开花关键基因的表达量。结果显示:(1)ECT6基因组包含5个外显子和4个内含子,ECT6突变体分别插入在ECT6基因的第3和第5外显子;ECT7基因组包含6个外显子和5个内含子,ECT7突变体分别插入在ECT7基因的第4和第6外显子。(2)突变体ECT6和ECT7均为完全敲除的功能缺失突变体,在长日照下二者均表现出开花提前和叶片数减少。(3)在长日照和短日照下ect6 ect7双突变体均表现出开花提前和叶片数减少。(4)开花抑制基因FLC和成花素基因FTLC是关键的开花调控因子,qRTPCR分析结果显示,在ECT6 ECT7双突变体中, FLC的表达水平降低, FTLC的表达水平升高,且春化通路基因VIN3、VRN2、VRN5和自主通路基因FVE的表达水平也显著升高。(5)ECT6和ECT7均定位于细胞核和细胞质中。

    Abstract:

    m6A modification is one of the most abundant modifications on mRNAs and regulates the fate decisions of mRNAs. YT521B homology (YTH) domaincontaining proteins are typical m6A “readers” that recognize and bind m6A for posttranscriptional regulation of genes. To investigate the functions of Arabidopsis YTH domaincontaining proteins ECT6 and ECT7 and their molecular mechanisms, we conducted genotyping, semiquantitative PCR identification and flowering phenotype observation of ECT6, ECT7 and ECT6 ECT7 double mutants. Meanwhile, we also analyzed the subcellular localization of ECT6 and ECT7 by cytological observation and detected the expression of key flowering genes by qRTPCR. The results showed that: (1) ECT6 contains five exons and four introns, and the ECT6 mutants is inserted in exons 3 and 5 of the ECT6, respectively; the ECT7 contains six exons and five introns, and the ECT7 mutants is inserted in exons 4 and 6 of the ECT7, respectively. (2) Mutants ECT6 and ECT7 were both lossoffunction mutants, and they both exhibited earlier flowering and reduced leaf number under longday condition. (3) The ECT6 ECT7 double mutant showed earlier flowering and reduced leaf number under both longday and shortday conditions. (4) FLC and FTLC are key flowering regulators. qRTPCR results showed that the expression levels of the flowering repressor gene FLC were reduced and the expression levels of the florigenin gene FTLC were increased in the ECT6 ECT7 double mutants. In addition, the expression levels of the vernalization pathway genes VIN3, VRN2, VRN5 and the autonomous pathway gene FVE were also significantly increased. (5) Both ECT6 and ECT7 were localized in the nucleus and cytoplasm.

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刘丹阳,徐遵涛,丁 勇.ECT6和ECT7调控拟南芥开花时间的研究[J].西北植物学报,2022,42(10):1621-1628

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  • 在线发布日期: 2022-11-10
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