毛竹PeAMT2;1基因克隆及表达分析

Cloning and Expression Analysis of PeAMT 2;1 from Phyllostachys edulis

铵态氮是植物吸收利用的主要氮源之一,铵态氮的吸收主要通过铵转运蛋白(AMT)进行。为从分子水平上探讨竹子中AMT的分子特征,本研究通过RT-PCR的方法对毛竹(Phyllostachys edulis)中的一个AMT基因编码区进行了克隆分析,并对其系统进化和表达模式进行了分析。结果表明,该基因的开放阅读框为1458 bp,与NCBI数据库中毛竹序列(FP095543)的一致性为99.04%,被命名为PeAMT2;1。PeAMT2;1编码485个氨基酸,分子量为51.62 kDa。蛋白序列分析表明,PeAMT2;1编码的蛋白序列具有11个跨膜结构域,与其他植物的AMT2亚家族具有较高的同源性,聚类分析与水稻、二穗短柄草等的AMT2亚家族成员在同一个分支。组织表达分析表明,PeAMT2;1在毛竹的根、茎、叶中均表达,但在根和茎中表达量相对较高。本研究对于今后深入了解PeAMT2;1的基因功能奠定了基础。

Ammonium nitrogen(NH■) is one of the main nitrogen sources for plant absorption and utilization. The absorption of NH■ is mainly through the ammonium transporter protein(AMT). In order to explore the molecular characteristics of AMT in bamboo at the molecular level, the open reading frame(ORF) of one AMT gene in Phyllostachys edulis is cloned by RT-PCR, and the phylogenetic and expression patterns of this isolated gene are analyzed. The result shows that the ORF is 1456 bp, which has 99.04% identities with the bamboo sequence of FP095543 in the NCBI database, and it is named as PeAMT2;1. PeAMT2;1 encodes a protein of 485 amino acids with a molecular weight of 51.62 kDa, which has 11 transmembrane domains and higher homology with AMT2 subfamily from other plants. Phylogenetic analysis indicates that PeAMT2;1 is clustered in the clade with those members of ATM2 subfamily from Oryza sativa and Brachypodium distachyum. Tissue specific expression analysis shows that PeAMT2;1 is constitutively expressed in roots, stems and leaves of P. edulis, with relatively high level in roots and stems. This study lays a foundation for the in-depth understanding of PeAMT2;1 function in the future.