摘要： 为研究野大豆TIFY家族基因在野大豆耐盐碱过程中的分子特性及表达特性，以极度耐盐碱野大豆G07256的cDNA为模板，结合前期转录组数据，采用同源克隆的方法，获得了GsTIFY6B基因，其全长CDS大小为1 047 bp。GsTIFY6B蛋白编码348个氨基酸，分子量为36.8 ku，等电点为9.12，是一个不稳定蛋白。系统进化树分析结果显示GsTIFY6B与大豆、绿豆、木豆和蔓花生的TIFY家族蛋白同源关系最近，含保守域TIFY和Jas。采用荧光定量PCR技术检测GsTIFY6B在野大豆不同部位以及盐碱胁迫和激素处理下的转录水平，结果表明，GsTIFY6B在根中相对表达量最高；在盐胁迫处理下，GsTIFY6B在野大豆根和叶中的表达量均表现出上调；在碱胁迫处理下，GsTIFY6B在野大豆叶和根中的表达量表现出先下调后上调表达，推测该基因可能参与野大豆盐碱胁迫防御反应；在ABA胁迫处理下，GsTIFY6B在野大豆的根中表现出下调表达，而在叶中6，12 h出现上调表达；在MeJA胁迫处理下，GsTIFY6B在根中先下调后上调表达，而在叶中表现出上调。研究表明，GsTIFY6B可能通过参与ABA和MeJA信号通路积极响应盐碱胁迫，对后期研究野大豆耐盐碱的分子机制提供了理论基础。

关键词: 野大豆, GsTIFY6B, 基因克隆, 盐碱胁迫

Abstract: In order to determine the molecular characterization and expression patterns of TIFY family genes involved in saline and alkali stress in Glycine soja, according to previous transcriptome data, GsTIFY6B was obtained by homologous cloning using cDNA of Glycine soja G07256, with extremely tolerant ability to saline and alkali stress, as its template. The full-length CDS was 1 047 bp. GsTIFY6B protein was an unstable protein with 348 amino acids. Its molecular weight was 36.8 ku and its isoelectric point was 9.12. Phylogenetic tree analysis showed the closest homology of GsTIFY6B to the TIFY family proteins of GmTIFY6B, VrTIFY6B, CcTIFY6B and AdTIFY6A. All those proteins had the conserved domains TIFY and Jas. The transcription level of GsTIFY6B in different parts of Glycine soja as well as response to saline and alkali stress and hormone treatment was detected by Real-time fluorescence quantitative PCR. The results showed that the highest relative expression level of GsTIFY6B in roots. Under salt stress, the expression of GsTIFY6B in roots and leaves showed an up-regulation.Under alkali stress, the expression of GsTIFY6B in roots and leaves showed the first down-regulation and then up-regulation expression. It is speculated that the gene might be involved in the defense reaction of saline-alkali stress.Under the ABA treatment, GsTIFY6B showed down-regulated expression in the roots and up-regulated expression at 6, 12 h in leaves. Under MeJA stress, GsTIFY6B expression was decreased first and then increased in roots, but showed up-regulation in leaves. This study suggested that GsTIFY6B could positively respond to saline and alkali stress by participating in the ABA and MeJA signaling pathways. These results will provide a theoretical basis for the later study on the molecular mechanism of saline-alkali-tolerant response of Glycine soja.

Key words: Glycine soja, GsTIFY6B, Gene cloning, Saline and alkali stress

中图分类号: 

• Q785

• S565.03