油棕硼转运通道蛋白基因NIP5的克隆及其在缺硼下的表达分析

Cloning and Expression Analysis of EgNIP5;1 Gene in Oil Palm Under Boron Deficiency

为了研究油棕硼转运的机制,以期为后续硼高效油棕品种筛选提供理论依据。以功能明确的拟南芥AtNIP5;1序列为参考,克隆了油棕硼转运通道蛋白EgNIP5;1基因的全长序列,并对其基因结构、序列特征、进化关系、跨膜结构和表达模式进行分析。EgNIP5;1的开放阅读框长度为894 bp,编码297个氨基酸;EgNIP5;1具有NIP保守的NPS、NPV和Ar/R位点,包含4个外显子3个内含子,具有5个跨膜结构。进化分析发现EgNIP5;1与可可TcNIP5的亲缘关系较近。荧光定量PCR分析发现EgNIP5;1主要在油棕根部表达,其表达量受到低硼的诱导,在缺硼处理48 h达到最大值,是对照的8.2倍。该实验表明EgNIP5;1可能参与油棕缺硼的响应,在硼的吸收中起着重要的作用。

This research aims to investigate the mechanism of boron transportation in oil palm and lay a theoretical foundation for the subsequent screening of boron efficient varieties. According to the sequence of AtNIP5;1 which is functional verified, we cloned the full-length sequence of boron transport channel protein gene EgNIP5;1. The gene structure, sequence characteristics, phylogenetic relationships, transmembrane domain and expression pattern were analyzed. The length of open reading frame of EgNIP5;1 was 894 bp,which coded a protein of 297 amino acids. EgNIP5;1 had highly conserved NPS, NPV and Ar/R motifs. EgNIP5;1 contained 4 extrons and 3 introns, and 5 transmembrane structures. Phylogenetic analysis showed that EgNIP5;1 was closely related to TcNIP5 of cacao. Real-time fluorescence quantitative PCR analysis showed that EgNIP5;1 was mainly expressed in roots of oil palm, and the expression ofEgNIP5;1 was induced by boron deficiency. The expression of EgNIP5;1 reached the maximum level at 48 h after boron deficiency treatment,which was 8.2 times of the control. This research suggests that EgNIP5;1 is possibly involved in the response to boron deficiency and plays an important role in boron absorption of oil palm.