摘　要：为鉴定可用于大豆（Glycine max）遗传改良的优异基因源，从大豆基因组中鉴定获得一个大豆GmWIN1-6转录因子，应用生物信息学工具和实时荧光定量反转录PCR（qRT-PCR）技术系统分析GmWIN1-6转录因子的理化性质、蛋白结构、时空表达谱及盐胁迫响应。结果表明：从大豆花组织中克隆到GmWIN1-6基因的开放阅读框（ORF），其编码蛋白由176个氨基酸组成，在N端具有一个保守的AP2结构域，属于亲水性蛋白；GmWIN1-6蛋白的二级结构中，无规则卷曲占比例最大；三级结构与拟南芥（Arabidopsis thaniana）AtWIN1相似；系统进化分析显示GmWIN1-6与AtWIN1亲缘关系最近；GmWIN1-6转录因子在大豆各组织中的表达差异明显，其中在花组织中表达量最高，其次为种子发育中后期，且与种子油脂积累时期基本吻合；在大豆幼苗盐胁迫条件下，GmWIN1-6基因上调表达。这些试验发现预示着GmWIN1-6转录因子可能参与大豆种子发育和油脂富集以及幼苗胁迫响应的调控，为大豆遗传改良和分子育种提供理论依据。
关键词：大豆；WIN1转录因子；油脂积累；盐胁迫；表达分析
中图分类号：S565.1　　　　　　文献标志码：ADOI：10.3969/j.issn.1007-7146.2021.02.009

Abstract: In order to identify an excellent gene source that can be used for soybean genetic improvement, a soybean GmWIN1-6 transcription factor was obtained from soybean (Glycine max) genome identification. Using bioinformatics tools and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) technology, the physical and chemical properties, protein structure, temporal and spatial expression profile and salt stress of GmWIN1-6 transcription factor were systematically analyzed. The results showed that the open reading frame (ORF) of GmWIN1-6 gene was cloned from soybean flower tissue, its encoded protein consists of 176 amino acids, has a conserved AP2 domain at the N-terminus, and is a hydrophilic protein. In the secondary structure of GmWIN1-6 protein, random coils account for the largest proportion. The tertiary structure is similar to AtWIN1 of Arabidopsis (Arabidopsis thaniana). Phylogenetic analysis showed that GmWIN1-6 is closely related to AtWIN1. The expression of GmWIN1-6 transcription factors in soybean tissues is significantly different. Among them, the expression level is the highest in flower tissues, followed by higher expression levels in the middle and late stages of seed development, and basically coincides with the accumulation period of seed oil. Under salt stress in soybean seedlings, GmWIN1-6 gene up-regulated expression. These experimental findings indicate that the GmWIN1-6 transcription factor may be involved in the regulation of soybean seed development and oil accumulation and seedling stress response, and provide a theoretical basis for soybean genetic improvement and molecular breeding.
Key words: soybean; WIN1 transcription factor; oil accumulation; salt stress; expression analysis
（Acta Laser Biology Sinica, 2021, 30(2): 154-162）