玉米磷转运蛋白基因ZmPht3；1的克隆和功能分析

摘要/Abstract

摘要： Pht3（phosphate transporter 3）磷转运子家族属于一类低亲和力磷转运蛋白，在调节植株体内磷素的动态平衡中发挥重要作用。为了初步探讨玉米中ZmPht3；1基因的结构特征及其磷饥饿的响应机制，利用同源克隆的方法从耐低磷玉米自交系Mo17中分离得到ZmPht3；1基因，并运用实时荧光定量PCR和亚细胞定位的方法对其进行深入研究。结果表明，ZmPht3；1的编码区全长1 101 bp，编码366个氨基酸，含有典型的线粒体转运家族（mitochondrial carrier family，MCF）结构特征与6个疏水跨膜结构。荧光定量PCR分析表明，该基因在两个极端材料的根系与叶片中均有表达，而表达模式差异显著，在耐低磷玉米自交系Mo17的根系和叶片中表现为缺磷胁迫前期的一般性反应和后期的特异性反应。转化烟草的亚细胞定位结果显示，ZmPht3；1主要分布于细胞膜上，可能是一个双亲和转运体，在玉米响应磷饥饿胁迫过程中发挥重要的适应性调节作用。

关键词: 玉米, 磷转运蛋白, 基因表达, 亚细胞定位

Abstract: Pht3（phosphate transporter 3）family belonging to low-affinity phosphate transporters play an important role in the regulation of phosphorus homeostasis in plants. In order to explore the characterization of gene structure and response mechanism of ZmPht3；1 to phosphorus（P）starvation，the gene ZmPht3；1 was isolated from low-phosphorus（low-P）tolerance maize inbred line Mo17 by homology cloning. Then the quantitative real-time PCR and subcellular localization of ZmPht3；1 were performed for further research in this study. The results showed that the complete CDS of ZmPht3；1 was 1 101 bp encoding 366 putative amino acids in length，and contained 6 hydrophobic transmembrane domains and had a typical conservation structure domain of mitochondrial carrier family. The PCR results exhibited that the gene expressed in roots and leaves，and the expression patterns were significantly different in the two extreme materials，respectively. While gene expression pattern in low-P tolerance maize inbred line Mo17 both roots and leaves revealed general and special response to P deficiency stress in early and late stages，respectively. Subcellular localization in the transformed tobacco（Nicotiana benthamiana）showed that ZmPht3；1 mainly expressed in cytoplasm membrane and it was a phosphate transporter in both low- and high-P environment，and played a key role in the process of adapting to P starvation in maize.

Key words: maize, phosphate transporters, gene expression, subcellular localization

中图分类号:

10.13560/j.cnki.biotech.bull.1985.2016.12.012

吴珊,林丹. 玉米磷转运蛋白基因ZmPht3；1的克隆和功能分析[J]. 生物技术通报, 2016, 32(12): 65-71.

WU Shan,LIN Dan. Cloning and Characterization of ZmPht3；1，a Phosphate Transporter Gene in Maize[J]. Biotechnology Bulletin, 2016, 32(12): 65-71.

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