金花茶CnMYB4转录因子的克隆及表达分析

Cloning and Expression Analysis of CnMYB4 Transcription Factor of Camellia Nitidissima

  • 摘要: 为解析金花茶CnMYB4转录因子在金花茶花瓣呈色以及类黄酮合成中的作用,以金花茶花瓣作为研究材料,克隆了金花茶CnMYB4转录因子的全长序列,随后对其进行生物信息学分析,并通过荧光定量PCR技术(qRT-PCR)分析了CnMYB4转录因子在根、茎、叶、盛开的花等金花茶不同组织中的时空表达模式。结果表明,金花茶CnMYB4的开放阅读框长度为735 bp,共编码244个氨基酸,该蛋白具有Myb_DNA-binding、SANT等保守结构域,以及C1(RGIDPxTHRPL/INE)、C2(PDLNLD/ELxIG/S)、Zf(GY/FDFLGL)、C4(FLGLX4-7V/LLD/GF/YR/SX1LEMK)等保守基序。系统进化分析发现,金花茶CnMYB4蛋白与茶树CsMYB4a蛋白的亲缘关系最近,它们被聚类于同一个小分支中。qRT-PCR分析结果表明,CnMYB4在金花茶花中的表达量相对较高,在叶片中的表达量相对较低。在金花茶花开放过程中,CnMYB4转录因子的表达量在幼蕾期相对较高,在半开期、盛开期相对较低。

     

    Abstract: To investigate the role of the CnMYB4 transcription factor in flower color and flavonoid synthesis in Camellia sinensis, the full-length sequence of CnMYB4 transcription factor as research materials was cloned. Subsequently, bioinformatics analysis was conducted. The temporal and spatial expression patterns of CnMYB4 in different tissues of Camellia sinensis were analyzed using quantitative fluorescence PCR (qRT-PCR). The results revealed that the open reading frame length of CnMYB4 was 244 amino acids. The protein contained conserved domains such as Myb_DNA binding and SANT, along with C1 (RGIDPxTHRPL/INE), C2 (PDLNLj/E LxIG/S), governmentY/F DFLGL(G), C4 (FLGLX47V/LLj/GF/YR/SX1LEMK), and conservative Motif. Phylogenetic analysis demonstrated a close relationship between the CnMYB4 protein from Camellia sinensis and CsMYB4 a protein from camellia camellia, clustering them within the same small clade. qRT-PCR analysis indicated that the expression level of CnMYB4 was relatively high in flowers but relatively low in leaves of Camellia sinensis. During opening stages, expression levels were highest during young bud stage but decreased during half-opening stage and blooming stage.

     

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