滇水金凤TT8转录因子的克隆及表达分析
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西南林业大学

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国家自然科学基金项目(面上项目,重点项目,重大项目)国家自然科学基金 (32060364, 32060366, 31860230);云南省重大科技专项 (202102AE090052);云南省教育厅科学研究基金项目(2022Y611);园林植物遗传改良与高效繁育博士生导师团队 (503210103)


Cloning and Expression Analysis of Transcription Fctor TT8 in Impatiens uliginosa
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The National Natural Science Foundation of China (32060364, 32060366, 31860230),Major Science and Technology Projects in Yunnan Province (202102AE090052),Yunnan Provincial Department of Education Science Research Fund Project(2022Y611),Doctoral Tutor Team for Genetic Improvement and High-efcient Propagation of Landscape Plants in Yunnan Province(503210103).

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    摘要:

    TT8(TRANSPARENT TESTA 8)基因是bHLH(basic helix-loop-helix)家族的重要成员之一,其在花青素合成中发挥着重要作用。为探究TT8基因对滇水金凤花色的影响,本研究以滇水金凤花器官为材料,通过RT-PCR等技术克隆IuTT8基因,并对其进行生物信息学;利用qRT-PCR分析该基因在不同花色和不同花发育阶段的表达模式。结果表明:(1)成功克隆得到滇水金凤IuTT8基因,其编码区全长为2 136 bp,编码712 aa,为亲水性不稳定蛋白,gDNA全长为3 938 bp,共有6个内含子;结构域分析发现该蛋白属于bHLH超家族成员,与喜马拉雅凤仙花、山茶等物种的TT8蛋白同源且Motif基序相似。(2)IuTT8与同属植物喜马拉雅凤仙花聚在一支,相似性约86.34%;多序列比对和系统进化分析显示TT8蛋白的结构域高度保守。(3)IuTT8基因在4种不同花色滇水金凤及其4个不同发育阶段均有表达,除白色外,其表达量均随花发育的进行呈先升后降的趋势;且IuTT8基因的表达量与花色呈正相关,其中以深红色表达量最高,白色表达量最低,深红色 S3 的表达量约为白色 S2 时期的48倍。研究表明滇水金凤IuTT8基因与花色正相关,推测其在滇水金凤花色的调控中发挥重要作用。

    Abstract:

    The TT8 (TRANSPARENT TESTA 8) gene is an important member of the bHLH (basic helix-loop-helix) family, playing a crucial role in anthocyanin synthesis. In order to explore the effect of TT8 gene on the color of Impatiens uliginosa, the IuTT8 gene was cloned by RT-PCR from Impatiens uliginosa, and its bioinformatics was carried out, qRT-PCR was used to analyze the expression patterns of the gene in different flower colors and different flower development stages. The results showed that: (1) The IuTT8 gene of I. uliginosa was successfully cloned, and its coding region was 2 136 bp, encoding 712 aa, which was a hydrophilic unstable protein. The gDNA was 3 938 bp, with 6 introns. Structural domain analysis showed that the protein belonged to the bHLH superfamily and was homologous to the TT8 proteins of Impatiens glandulifera, Camellia sinensis var. assamica and other species, and they have similar Motifs. (2) IuTT8 clustered with Impatiens glandulifera, and the similarity is about 86.34%; multi-sequence alignment and phylogenetic analysis showed that the domain of TT8 protein was highly conserved. (3) IuTT8 gene was expressed in I. uliginosa with 4 different flower colors and at 4 different stages of flower development. Except for white, the expression level of IuTT8 showed an upward trend followed by a downward trend with the progress of flower development. The expression level of IuTT8 was positively correlated with flower color, with the highest expression level in deep-red flowers and the lowest in the white-flowers. The expression level of IuTT8 in the S3 period of deep-red flower was approximately 48 times higher than that in the S2 period of white flower. The results showed that IuTT8 gene of I. uliginosa is a transcription factor of bHLH family, which was positively correlated with flower color, suggesting that it may play an important role in the regulation of flower color in I. uliginosa.

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李新艺,李洋,向南星,等.滇水金凤TT8转录因子的克隆及表达分析[J].西北植物学报,2023,43(11):1813-1819

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  • 收稿日期:2023-05-25
  • 最后修改日期:2023-07-26
  • 录用日期:2023-08-03
  • 在线发布日期: 2023-11-14
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