小麦转录因子TabHLH041基因的克隆与表达分析

Cloning and Expression Analysis of Transcriptional Factor Gene Tab HLH041 from Wheat

利用RT-PCR方法从小麦品种辽春10号中克隆得到1个b HLH转录因子基因Tab HLH041,并对其表达特性进行分析,以期为研究b HLH转录因子在小麦发育中的功能奠定基础。结果表明,Tab HLH041c DNA开放阅读框(ORF)长1 179 bp,编码392个氨基酸,Tab HLH041蛋白在C端含有1个b HLH结构域;该基因包括4个外显子和3个内含子。系统发育分析发现,Tab HLH041蛋白与大麦b HLH蛋白亲缘关系最近。实时荧光定量PCR分析表明,总体上该基因的表达具有昼夜节律性,在光照条件下表达量呈下降趋势,在黑暗条件下表达量呈上升趋势。在低温条件下,Tab HLH041基因的表达呈现前期被诱导、后期被抑制的特征,处理24 h时低温的抑制作用最明显;在高盐、外源ABA和干旱处理条件下,Tab HLH041基因的表达持续受到抑制。综上,Tab HLH041基因可能参与光周期及逆境胁迫的信号转导途径。

In order to lay a foundation for the study of the function of b HLH transcription factors in the development of wheat,a b HLH transcription factor Tab HLH041 gene was cloned by RT-PCR method from wheat variety Liaochun No. 10,and its expression pattern was analyzed. The results showed that the open reading frame of Tab HLH041 was 1 179 bp in length that encoded 392 amino acids. The Tab HLH041 protein contained a b HLH domain in its C terminal. The analysis of the sequence of Tab HLH041 from genome DNA showed that Tab HLH041 gene contained four exons and three introns. Phylogenetic analysis showed that the sequence of Tab HLH041 was closest to Hordeum vulgare b HLH protein. Real time PCR analysis revealed that the expression of Tab HLH041 displayed the day-night rhythm,and its expression level decreased in the light while increased in the dark. The expression of Tab HLH041 was induced in the early time,and then was inhibited under the treatment of low temperature,the inhibition was very significant when treated for 24 h. The expression of Tab HLH041 was inhibited continuously under the treatment of high salt,ABA and PEG. In summary,the results suggest that Tab HLH041 may be involved in the signal transduction pathway of photoperiod and adversity stress.