小麦分蘖角度TaTAC1基因同源克隆及表达分析
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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)


Molecular characterization and expression analysis of TaTAC1 gene in Triticum aestivum L.
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The National Natural Science Foundation of China (General Program, Key Program, Major Research Plan)

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    摘要:

    分蘖角度影响植物群体结构、光合效率以及植株的形态建成,最终影响其产量及品质,小麦中有关分蘖角度的研究罕见报道。为解析TaTAC1的表达模式,初步了解该基因的分子遗传机制及其与分蘖角度的遗传关系。该研究以CN16、SM969、Lan2399以及SHW-1为材料,利用同源克隆分离TaTAC1;利用生物信息学软件对TaTAC1进行序列特征分析;应用实时荧光定量PCR对其表达模式进行分析;并将TaTAC1蛋白进行亚细胞定位。TaTAC1长度约1.1~1.2kb,包括780bp完整的ORF和320~370bp的3’-UTR,ORF编码260个氨基酸。TaTAC1的cDNA序列分为两类,其中CN16-2、SM969-2的第10号碱基发生突变,引起终止子提前,导致TaTAC1表达受阻;Lan2399-2和SHW-1-2在109-115碱基位置有“CGCGCG”片段插入导致蛋白质β-折叠片减少。实时荧光定量PCR分析表明,TaTAC1在分蘖期的叶鞘、茎高效表达,分蘖节其次,叶与根表达量最低;SPSS20.0分析该基因在分蘖节各时期表达量与分蘖角度显著正相关,Pearson相关性系数为0.677;其他组织表达量与分蘖角度无显著相关性。TaTAC1亚细胞定位于细胞膜。TaTAC1在不同时期下不同组织均有表达,其具有组织时空表达特异性;该基因在叶鞘、茎、分蘖节表达高,分蘖节处表达量与表型显著相关且蛋白定位于细胞膜,推测TaTAC1在mRNA水平上正向调控分蘖角度,且其可能参与生长素极性运输过程从而改变分蘖角度大小。

    Abstract:

    Tillering angle affects plant population structure, photosynthetic efficiency and morphogenesis, ultimately affects the yield and quality. Studies of tillering angle had rare reports in wheat. For parsing the expression pattern of TaTAC1 and having a preliminary understanding of the molecular genetic mechanism and genetic relationship with the tillering angle,This study used CN16,SM969,Lan2399,SHW-1 as materials and homology-based cloning to separate TaTAC1,using bioinformatics software to analyze the characteristic of TaTAC1 sequence, applying real-time fluorescent quantitative PCR to analyze its expression pattern. In order to investigate the function of TaTAC1,this study used subcellular localization by transient expression of TaTAC1-GFP fusion protein on onion epiderm.The length of TaTAC1 is about 1.1~1.2 kb, including 780 bp complete ORF and 320~370 bp 3 'UTR. ORF encoded 260 amino acids. There were two types of TaTAC1 cDNA sequence. there was a base mutation at No.10 of the cDNA sequence in CN16-2 and SM969-2,which causes to terminate in advance so as to gene expression being block. In addition, there were 6 bases insert “CGCGCG” in 109-115 base location that led to protein β-pleated sheet decreasing. Real-time fluorescent quantitative PCR analysis showed that TaTAC1 in leaf sheath, stem had efficient expression of tillering stage, tillering node, leaf and root expression quantity minimum.SPSS20.0 analyzed the gene expression in tillering node of each period and tillering angle had significant positive correlation. Pearson correlation coefficient was 0.677;Other organizations expression quantity and tillering angle had no significant correlation.TaTAC1 had subcellular localization in the cell membrane.TaTAC1 was expressed under different organization in different periods .It expressesd specificity organize time and space. The gene expressed in leaf sheath, stem, tillering node efficiently. Location expression quantity of tillering node was associated with a significant phenotypic and protein in the cell membrane. Thus speculate TaTAC1 had positive regulation with tillering angle at the mRNA level, and it may be involved in auxin polar transport process to change the size of tillering angle.

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曹鑫,邓梅,张正丽,等.小麦分蘖角度TaTAC1基因同源克隆及表达分析[J].植物遗传资源学报,2017,18(1):125-132.

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  • 收稿日期:2016-02-13
  • 最后修改日期:2016-05-06
  • 录用日期:2016-07-01
  • 在线发布日期: 2017-01-17
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