小麦转录因子基因TaERF7的克隆及其表达分析
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Cloning and Expression Analysis of Transcription Factor Gene TaERF7 in Wheat (Triticum aestivum)
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    摘要:

    温光敏核不育小麦(Triticum aestivum)百农不育系(Bainong sterility,BNS)是一种良好的小麦杂种优势利用材料,利用其低温不育、高温可育的特性可以实现 “两系法”杂交小麦育种。该研究为找到BNS育性转换的关键基因,从已有BNS不育和可育花药的基因芯片数据中,鉴定得到表达存在差异的TaERF7基因,并通过设计引物克隆了TaERF7的cDNA和启动子序列,采用qRTPCR分析TaERF7对不同温度和光照的响应。结果显示:(1)生物信息学分析表明,TaERF7基因的CDS区有660 bp,编码219个氨基酸; TaERF7蛋白含有AP2结构域和2个EAR基序,属于第二类乙烯响应因子(Ethylene response factor,ERF); TaERF7的氨基酸序列与拟南芥(Arabidopsis thaliana)AtERF4同源,可能是一种转录抑制因子; TaERF7启动子区含有多个光响应和低温响应的顺式作用元件。(2)qRTPCR结果表明,TaERF7在BNS的不同组织与器官中均有表达;在长日照(14 h)下,TaERF7在BNS中的表达量下调约0.47倍,而在短日照(10 h)处理下,TaERF7在BNS中的表达量上调约1.14倍;4 ℃的低温处理使TaERF7表达量在2 h内上调了约25.7倍,并且在48 h内一直保持较高的水平,而在37 ℃下虽然可以使TaERF7表达量在1 h内上调约0.71倍,但2 h后便急剧下调,到12 h时其表达量与对照相比已下调了约0.85倍。该研究结果初步证明,TaERF7基因可能特异性结合下游基因启动子的GCC box、DRE和CRT元件,调控下游基因的表达,从而影响了BNS的育性。

    Abstract:

    BNS is an efficient thermophotosensitive genic male sterile line of wheat (Triticum aestivum). It is sterile in low temperature and fertile in high temperature. With these features, BNS was widely used in hybrid wheat breeding. In this study, based on the gene chip data of the anthers of BNS sterile line and fertile line, we found that the transcription factor TaERF7 was differentially expressed between the two lines, and cloned the cDNA and promoter sequences of TaERF7,analyzed TaERF7 response to temperature and day length by qRTPCR. The results showed that: (1) bioinformatics analysis revealed that the coding sequence (CDS) of TaERF7 is 660 bp and encoding of 219 amino acids. TaERF7 protein contains AP2 domain and two EAR motifs. It belongs to class Ⅱ ethylene response factors (ERFs). The amino acid sequence of TaERF7 is homologous to AtERF4 in Arabidopsis (Arabidopsis thaliana), which is a transcriptional repressor. The promoter region contains multiple cisacting elements with light and low temperature response. (2) The qRTPCR results showed that TaERF7 expressed in different tissues of BNS. The expression of TaERF7 was downregulated 0.47 fold under long daylight (14 h), while it was upregulated 1.14 fold under short day (10 h). The low temperature treatment at 4 ℃ increased the expression of TaERF7 about 25.7 fold in 2 h, and maintained a high level upto 48 h. The high temperature treatment at 37 ℃ increased the expression of TaERF7 about 0.71 fold in 1 h, but it was sharply downregulated to 0.85 fold after 2 h. According to the above results, we speculated that TaERF7 combined to cisacting element of downstream genes, regulated their expression, and caused the male sterility in BNS.

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李紫良,张建朝,李 政,等.小麦转录因子基因TaERF7的克隆及其表达分析[J].西北植物学报,2020,40(2):210-217

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  • 在线发布日期: 2020-05-02
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