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欧洲白榆UlGLK基因克隆及组织表达特异性研究

Gene Cloning and Specific Tissue Expression of UlGLK from Ulmus laevis

  • 摘要: GLK转录因子是GARP转录因子超家族成员,具有参与调控植物叶绿体发育、生物胁迫和非生物胁迫、植物衰老和激素信号转导等功能,该基因在叶色改良方面已成功应用于白桦(Betula platyphylla)及银中杨(Populus alba×P.berolinensis)等林木。为了探讨欧洲白榆(Ulmus laevis)UlGLK基因的功能,为后续白榆的分子设计育种提供参考,以欧洲白榆为研究材料,开展UlGLK基因克隆及序列分析,筛选qRT-PCR分析用的内参基因,分析UlGLK基因的组织表达特性。研究结果显示,UlGLK基因的cDNA长度为1 353 bp,编码450个氨基酸,与大麻(Cannabis sativa)、川桑(Morus notabilis)的同源性高。以白榆根、茎和叶组织cDNA为模板,根据Ubiquitin、Actin、Tubulin基因在上述3种组织中的表达水平,确定ubiquitin基因可作为后续qRT-PCR分析内参基因。UlGLK基因的组织表达特性分析显示,该基因在叶片组织中表达量最高,在根和茎组织中表达量较低。研究结果为欧洲白榆后续分子设计育种提供参考。

     

    Abstract: GLK transcription factor is a member of GARP transcription factor superfamily,which plays a role in regulating plant chloroplast development,biological and abiotic stress,senescence and hormone signal transduction. GLK had been successfully used in leaf color improvement of Betula platyphylla and Populus alba ×P. berolinensis,etc. In order to explore the function of UlGLK gene in Ulmus laevis and provide reference for molecular design and breeding of U. laevis. UlGLK was cloned from U. laevis and sequenced and the internal reference genes were screened for qRT-PCR analysis and the tissue expression characteristics of UlGLK gene was analyzed. The results showed that the ORF of UlGLK gene was 1 353 bp in length,encoded 450 amino acids,which had high homology with cannabis sativa and Morus notabilis. Based on the expression levels of Ubiquitin,Actin and Tubulin genes in U. laevis root,stem and leaf respectively,ubiquitin gene was identified as an internal reference for qRT PCR analysis. Analysis of tissue expression characteristics of UlGLK showed that the expression level of UlGLK was the highest in leaf,but lower in root and stem. The results provided a reference for further molecular breeding of U. laevis.

     

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