檀香小G蛋白Rac1基因的克隆与功能分析
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中国林科院基本科研业务费(CAFYBB2019QB003);


Cloning and Functional Analysis of SaRac1 Gene in Santalum album Linn.
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    摘要:

    植物Rac蛋白属于小分子G蛋白ROP家族,广泛参与活性氧(ROS)产生、激素信号转导和组织形态建成。檀香(Santalum album Linn.)是著名的珍贵树种,为半寄生植物,其正常生长需要根部特化的吸器从其他寄主植物摄取营养物质。该研究基于全长转录组数据,采用RTPCR方法克隆得到了1个檀香Rac基因,命名为SaRac1。结果表明:(1)SaRac1基因全长594 bp,编码197个氨基酸,分子量21.55 kD,理论等电点9.32,为亲水性蛋白。(2)进化分析显示,SaRac1蛋白和拟南芥(Arabidopsis thaliana)AtRac1~6、AtRac9和AtRac11蛋白同属于典型的植物RacⅠ家族蛋白。(3)结构预测显示,该蛋白在N端为保守的G结构域,蛋白C端具有CaaL保守基序。(4)原生质体亚细胞定位试验显示,SaRac1蛋白定位于细胞核和细胞质。(5)组织特异性表达分析显示,SaRac1基因在根和吸器中表达量最高,幼叶和茎中表达量较高,在成熟叶和老叶中表达量较低。(6)用寄生植物吸器诱导因子2, 6二甲氧基对苯醌(DMBQ)处理,发现SaRac1受到DMBQ的强烈诱导,表达量在4 h达到最高。研究推测,SaRac1基因受吸器诱导因子调控进而参与檀香吸器形成过程。

    Abstract:

    Rac proteins of plants belong to Rhorelated GTPase family (ROP family) and play important roles in reactive oxygen species (ROS) production, hormone signal transduction and tissue morphogenesis. Sandalwood (Santalum album Linn.) is a precious and semiparasitic plant with specialized root haustoria to absorb nutrients from other host plants. Based on the fulllength transcriptome data, a Rac protein gene was isolated from Santalum album by RTPCR and named SaRac1. The results showed that: (1) SaRac1 contains a 594 bp opening reading frame, encoding a putative hydrophilic protein of 197 amino acids with molecular weight 21.55 kD and isoelectric point 9.32. (2) Phylogenetic tree analysis showed that the amino acid sequence exhibits high similarity to AtRac1-6, AtRac9 and AtRac11 in Arabidopsis thaliana and belongs to typical Rac I proteins in plants. (3) Sequence alignment indicated the protein has conserved G domains in the Nterminal and CaaL motif in the Cterminal. (4) Subcellular localization analysis showed that SaRac1 localized in the nucleus and cytoplasm. (5) Tissuespecific expression analysis revealed that the transcript level of SaRac1 was higher in roots and haustoria than those in other issues. (6) SaRac1 was induced by 2, 6dimethoxyρbenzoquinoe (DMBQ, an important haustoriuminducing factor) and reached the highest transcript level at 4 h. These results indicate that SaRac1 might be induced by the chemical factor and involved in the development of haustorium.

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卞 展, 刘小金,洪 舟,等.檀香小G蛋白Rac1基因的克隆与功能分析[J].西北植物学报,2021,41(1):46-51

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  • 在线发布日期: 2021-02-26
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