马铃薯StZIP8–like基因的克隆及镉转运功能分析
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国家马铃薯产业技术体系(CARS–09–ES16)


Cloning of StZIP8-like gene in potato and analysis of its function in cadmium transport
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    摘要:

    为了探讨StZIP8–like基因的表达模式和功能,从马铃薯中克隆得到StZIP8–like基因的全长序列,通过生物信息学方法对该基因的结构、理化性质及亲缘关系进行预测;利用RT–qPCR技术,分析其在镉胁迫下的表达模式;构建融合表达载体,分析StZIP8–like蛋白在烟草叶片中的亚细胞定位;通过酵母异源表达试验,分析StZIP8–like的镉转运功能。结构分析结果表明,StZIP8–like序列全长1 044 bp,编码347个氨基酸,含有1个ZIP保守结构域及7个跨膜结构域;系统进化分析结果表明,StZIP8–like蛋白与番茄及烟草中的ZIP8蛋白的亲缘关系较近。亚细胞定位结果显示,StZIP8–like蛋白定位于细胞质膜;RT–qPCR结果表明,StZIP8–like在马铃薯幼苗的根中表达量较高,并且Cd胁迫诱导下StZIP8–like基因在马铃薯根、茎和叶中的表达量均上调;酵母试验结果显示,StZIP8–like过表达的酵母对Cd更加敏感,具有Cd吸收转运活性。以上结果说明StZIP8–like受到Cd胁迫的诱导表达,可能参与马铃薯吸收Cd的过程。

    Abstract:

    In order to explore the expression pattern and function of the StZIP8-like gene, the full-length sequence of the StZIP8-like gene was cloned from potato. The structure, physicochemical properties, and genetic relationship of this gene were predicted by bioinformatics methods. The expression pattern of this gene under cadmium(Cd) stress was analyzed using the RT-qPCR technique. A fusion expression vector was constructed to analyze the subcellular localization of the StZIP8-like protein in tobacco leaves. Through a yeast heterologous expression experiment, the Cd transport function of StZIP8-like was analyzed. The results of structural analysis showed that the full-length sequence of StZIP8-like was 1 044 bp, encoding 347 amino acids, containing a ZIP conserved domain and 7 transmembrane domains. Phylogenetic analysis revealed that the StZIP8-like protein had a close genetic relationship with the ZIP8 proteins in tomato and tobacco. The results of subcellular localization indicated that the StZIP8-like protein was localized in the plasma membrane. The RT-qPCR results showed that StZIP8-like was highly expressed in the roots of potato seedlings, and Cd stress induced the upregulation of the expression of the StZIP8-like gene in the roots, stems, and leaves of potato. The yeast experiment showed that the yeast with overexpression of StZIP8-like was more sensitive to Cd and had Cd absorption and transport activity. The above results indicate that StZIP8-like is induced by Cd stress and may be involved in the process of Cd absorption in potato.

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杨丽红,席银凯,王丹,胡新喜*.马铃薯StZIP8–like基因的克隆及镉转运功能分析[J].湖南农业大学学报(自然科学版),2025,51(3):16-23.

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  • 在线发布日期: 2025-07-15
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