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何庆梦,高 翔,杨明明,董 剑,李晓燕,赵万春.普通小麦 TaTFL1基因的克隆及表达特性研究[J].麦类作物学报,2018,(9):1020
普通小麦 TaTFL1基因的克隆及表达特性研究
Cloning and Expression Analysis of TaTFL1 Gene in Wheat(Triticum aestivum L.)
  
DOI:10.7606/j.issn.1009-1041.2018.09.02
中文关键词:  Wheat   TaTFL1  Gene cloning  Expression analysis
英文关键词:小麦   TaTF1  基因克隆  表达分析
基金项目:国家现代农业产业技术体系研究项目(CARS-3-2-47);“十二五”农村领域国家科技计划课题(2011AA100501)
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何庆梦,高 翔,杨明明,董 剑,李晓燕,赵万春 (1.西北农林科技大学农学院陕西杨凌 712100 2.陕西省小麦工程技术研究中心/陕西省小麦新品种培育工程研究中心陕西杨凌 712100) 
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中文摘要:
       TFL1 gene as the plant inflorescence meristem specific gene,its main function is to maintain the infinite vegetative growth of plant. In this study,gene cloning and reverse transcription PCR technology were used to obtain the CDS sequence of TaTFL1 gene from Chinese spring,and bioinformatics analysis was conducted to analyze the structure,function and sequence homology; finally,by using the quantitative real-time PCR(qRT-PCR) technique,the expression pattern of this gene was detected in different tissues at different developmental stages in common wheat. The result show that TaTFL1 is a hydrophobic protein,without signal peptide,and located in cytoplasm. The homology and phylogenetic tree analysis of TaTFL1 genes displayed close genetic relationship between wheat and monocotyledonous plants. We also analyzed the promoter and found a large number of regulatory elements related to light responsiveness,and the cis-acting regulatory elements related to meristem expression(CAT-box) exist in the promoter.qRF-PCR analysis showed that TaTFL1 was detected in various tissues. The expression level in leaves was very low,but the highest expression level was detected in the stem below spike. Expression level was significantly higher in both leaves and stem below spike before anthesis than that after anthesis; lower expression was detected in the bud,glume and grain; at different developmental stages of spike,the young spike(1-2 cm) has the highest expression level,and the expression level showed a downward trend with the development; young spike always maintained a high expression level compared with other organs. In summary, TaTFL1 has a typical structure of the TFL1 gene family,and we speculated that TaTFL1 may play an important role mainly at the early spike development stage.
英文摘要:
       TFL1基因是植物花序分生组织特异性基因,其主要功能是维持植物营养生长和花序的无限生长。本研究利用同源克隆技术获得了 TaTFL1部分同源基因的cDNA序列和启动子序列,并利用生物信息学技术对其序列组成、结构特征及进化关系进行了初步分析,并对启动子区顺式作用元件进行了预测,进一步通过实时荧光定量PCR(Quantitative real-time PCR,qRT-PCR)技术对该基因在中国春不同组织以及各组织不同发育阶段的表达情况进行分析。结果表明,TaTFL1蛋白为疏水性蛋白,无信号肽,定位于细胞质中;进化分析显示, TaTFL1与单子叶植物的亲缘关系较近;在启动子中存在着大量的光响应元件,以及分生组织表达相关的顺式调控元件(CAT-box)。qRT-PCR结果表明, TaTFL1基因在各组织中皆有表达,且均表现为花前表达量明显高于花后;在叶片中表达量较低,而在花前的穗下茎中表达量最高,在幼芽、颖壳和籽粒中也有微弱的表达;不同发育阶段的幼穗中, TaTFL1在1~2 cm的幼穗中表达量最高,随着幼穗的发育呈现出下降的趋势,且与其他组织相比始终维持着较高的表达水平。综上所述,TaTFL1蛋白具有TFL1家族的典型结构,在小麦幼穗发育初期发挥重要作用。
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