番茄 SlDOGL4 转录因子基因的克隆和表达分析

Cloning and expression analysis of tomato SlDOGL4 transcription factor gene

摘要

为了明确 SlDOGL4 基因在番茄中的耐盐作用，以栽培番茄 Ailsa Craig 为试验材料，克隆该基因并通过生物信息学方法分析其理化性质和表达特性。结果表明， SlDOGL4 基因 CDS 全长 660 bp, 编码 219 个氨基酸；保守结构域分析结果表明，该基因仅含有 1 个 DOG1 结构域，属于 bzip 转录因子中的 DOG1 家族成员；系统进化关系表明，SlDOGL4 与马铃薯的 StDOGL4 蛋白亲缘关系较近。组织表达谱结果表明， SlDOGL4 基因主要在根和破色期果实中表达。亚细胞定位结果表明，SlDOGL4 蛋白定位于细胞核。 SlDOGL4 基因启动子上有多种与非生物胁迫和激素响应相关的顺式元件。构建 Pro SlDOGL4 ::GUS 融合表达载体，遗传转化番茄检测启动子活性，GUS 组织化学染色结果表明， SlDOGL4 在根、茎、生长点、种子等组织中都有表达，表明该基因在番茄整个生长发育过程中发挥重要作用。通过公共数据库中转录组数据分析表明， SlDOGL4 基因的表达受到盐、干旱、冷和热胁迫不同程度的诱导。为了进一步验证 SlDOGL4 基因的功能，在盐处理后， SlDOGL4 基因的转录水平呈先升高后降低的趋势，且在盐处理 2 h 后达到最高转录水平。研究结果为探索 SlDOGL4 基因在番茄耐盐中的功能奠定了基础。

Abstract

In order to clarify the salt-tolerance effect of SlDOGL4 gene in tomato, the cultivated tomato Ailsa Craig was used as the experimental material to clone this gene and analyze its physicochemical properties and expression characteristics through bioinformatics methods. The results showed that the full length of CDS of SlDOGL4 gene was 660 bp, encoding 219 amino acids. Conserved domain analysis showed that the gene contained only one DOG1 domain, which belonged to the DOG1 family of bzip transcription factor. Phylogenetic relationship showed that SlDOGL4 was closely related to StDOGL4 protein in potato. The results of tissue expression profile showed that SlDOGL4 gene was mainly expressed in root and fruit at the broken color stage. Subcellular localization results showed that SlDOGL4 protein was located in the nucleus. SlDOGL4 gene promoter has a variety of cis-elements related to abiotic stress and hormone response. The Pro SlDOGL4 :: GUS fusion expression vector was constructed to genetically transform tomato to detect promoter activity. GUS histochemical staining showed that SlDOGL4 was expressed in roots, stems, growing points and seeds, indicating that SlDOGL4 played an important role in the whole growth and development process of tomato. Analysis of transcriptome data from public databases showed that the expression of SlDOGL4 gene was induced by salt, drought, cold and heat stress to varying degrees. In order to further verify the function of SlDOGL4 gene, the transcription level of SlDOGL4 gene showed a trend of first increasing and then decreasing, and reached the highest transcription level after 2 hours of salt treatment. These results laid a foundation for exploring the function of SlDOGL4 gene in tomato salt tolerance.

关键词

番茄 / SlDOGL4 / 非生物胁迫 / 亚细胞定位 / 转录水平

Key words

Tomato / SlDOGL4 / Abiotic stress / Subcellular localization / Transcription level

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基金资助

国家自然科学基金(NSFC31801880)

河南省重点研发与推广专项(222102110407)