普通小麦TaZIP29-7B基因的克隆及表达研究

金属离子在小麦生长过程中起重要作用,但土壤中金属离子含量过高,会对小麦生长造成损害。锌铁转运蛋白ZIP家族(ZRT,IRT-like protein,ZIP)广泛存在于植物中,参与多种金属离子的转运,也能提高植物的耐盐性。利用同源克隆技术获得 AtZTP29在小麦中的同源基因TaZIP29-7B,并获得其启动子序列,利用生物信息学技术对所获得的基因以及启动子序列进行初步分析,预测启动子顺式作用元件,通过实时荧光定量PCR(Quantitative real-time PCR,qRT-PCR)技术分析小麦 TaZIP29-7B基因在根和叶中在不同金属离子溶液处理下的表达情况,以研究该基因在小麦体内中对重金属转运的作用。结果表明:序列分析显示 TaZIP29-7B基因(登录号MK203894)编码277个氨基酸,TaZIP29-7B蛋白为疏水性蛋白,定位在细胞膜上,存在信号肽,属于分泌蛋白,第89～263位置间的结构功能域ZIP,属于典型的ZIP超级家族结构域,具有8个跨膜区;进化分析显示TaZIP29-7B与单子叶植物山羊草(Aegilops tauschii)ZIP29蛋白同源性最高;启动子中存在多种响应胁迫的顺式作用元件;qRT-PCR结果显示,小麦 TaZIP29-7B基因在外界重金属盐的变化下表达量有所变化。说明研究结果为进一步改良小麦抗盐性提供了参考。

Cloning and Expression Analysis of TaZIP29-7B Gene in Wheat(Triticum aestivum L.)

Metal ions play an important role in the growth of wheat, and high concentration of metal ions in soil will damage wheat growth. The zinc-iron transporter ZIP family (ZRT, IRT-like protein, ZIP) is widely distributed in plants, involved in the transport of various metal ions, and can also improve the salt tolerance of plants. In this study, homologous cloning technology was used to obtain the homologous gene TaZIP29-7B of AtZTP29 in wheat, and its promoter sequence was obtained. The obtained gene and promoter sequence were analyzed by bioinformatics. The cis-acting element was used to analyze the expression of the TaZIP29-7B gene in roots and leaves under different metal ion solutions by real-time quantitative real-time PCR (qRT-PCR) and explored the role of heavy metal transport in wheat. Sequence analysis showed that TaZIP29-7B gene (accession number MK203894) encodes 277 amino acids, TaZIP29-7B protein is a hydrophobic protein, which is located on the cell membrane, and there is a signal peptide, which is a secreted protein, and the structural domain between positions 89-263 ZIP, belonging to the typical ZIP superfamily domain, has 8 transmembrane regions; evolutionary analysis shows that TaZIP29-7B has the highest homology with the monocotyledonous Aegilops tauschii ZIP29 protein; there are multiple cis-acting elements in the promoter responding to stress. qRT-PCR results showed that the expression level of TaZIP29-7B gene changed under the change of heavy metal salt. The research results provide a theoretical basis for further improving the salt tolerance of wheat.