茄子SmWRKY4的克隆及耐冷性功能验证
摘要：
前期研究发现茄子SmWRKY4的表达受到冷信号抑制。进一步以‘成都墨茄’为材料克隆出SmWRKY4全长序列，分析结果显示其开放阅读框为1 518 bp，编码505个氨基酸，含有2个WRKY结构域，属于第Ⅰ类WRKY转录因子。SmWRKY4启动子含有茉莉酸甲酯和水杨酸等激素响应元件及缺氧和低温等逆境胁迫响应元件。系统进化树分析表明，SmWRKY4与番茄和马铃薯的同源蛋白在进化关系上更近。表达模式分析显示SmWRKY4能够响应脱落酸、水杨酸和茉莉酸甲酯等外源植物生长调节剂和冷胁迫，并在茄子茎和花药中高表达。敲除SmWRKY4的突变体能够增强光合作用，降低细胞通透性，提高耐冷性。推测SmWRKY4负调控茄子耐冷性。
关键词: 茄子, SmWRKY4, 耐冷性, 克隆, 功能验证, 光合作用, 细胞通透性, 激素
Abstract:
Previous study showed that the expression level of SmWRKY4 was inhibited by cold stress. The full-length sequence of SmWRKY4 was cloned from‘Chengdu Moqie’in this study. Analysis results revealed that the open reading frame of the SmWRKY4 gene was 1 518 bp，encoding 505 amino acids which contained two WRKY domains. Consequently，SmWRKY4 protein belonged to group Ⅰ WRKY transcription factor. SmWRKY4 promoter contained hormone-response elements including MeJA and SA and stress-response elements including oxygen deficit and low temperature. Phylogenetic tree analysis suggested that SmWRKY4 protein was more closely related to its homologous proteins in tomato and potato. Expression pattern analysis showed that SmWRKY4 could respond to ABA，SA，MeJA and cold stress，and was highly expressed in eggplant stem and anther. SmWRKY4 mutation could enhance photosynthesis，reduce cell permeability and improve cold tolerance. These results suggested that SmWRKY4 negatively regulated cold tolerance in eggplant.
Key words: eggplant, SmWRKY4, cold tolerance, gene cloning, functional verification, photosynthesis, cell permeability, hormones
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