甘薯IbbZIP22基因克隆与表达分析  

Cloning and Expression Analysis of IbbZIP22 Gene in Sweet Potato

作　　者：孟鑫 王庆美[2,3] 侯夫云[2,3] 李爱贤[2,3] 董顺旭[2,3] 周媛媛 秦桢[2,3] 张立明[3,4] Meng Xin;Wang Qingmei;Hou Fuyun;Li Aixian;Dong Shunxu;Zhou Yuanyuan;Qin Zhen;Zhang Liming(Agronomy College of Qingdao Agricultural University,Qingdao 266109,China;Crop Research Institute,Shandong Academy of Agricultural Sciences/Scientific Observation and Experimental Station of Tubers and Root Crops in Huang-Huai-Hai Region,Ministry of Agriculture and Rural Affairs,Jinan 250100,China;Shandong Academy of Agricultural Sciences,Jinan 250100,China;College of Life Sciences,Shandong Normal University,Jinan 250014,China)

机构地区：[1]青岛农业大学农学院,山东青岛266109 [2]山东省农业科学院作物研究所/农业农村部黄淮海薯类科学观测实验站,山东济南250100 [3]山东省农业科学院,山东济南250100 [4]山东师范大学生命科学学院,山东济南250014

出　　处：《山东农业科学》2023年第1期1-7,共7页Shandong Agricultural Sciences

基　　金：泰山产业领军人才工程专项(LJNY202113);国家甘薯产业技术体系项目(CARS-10-B06);山东省薯类产业技术体系项目(SDAIT-16-04);山东省农业良种工程项目(2020LZGC004);山东省农业科学院作物研究所学科团队建设项目(CXGC2018E01);山东省农业科学院农业科技创新工程项目(CXGC2022A14)。

摘　　要：本研究以济薯25为材料,克隆得到IbbZIP22基因。该基因编码区全长1 368 bp,编码455个氨基酸。序列和结构分析发现该基因编码的蛋白上有bZIP_plant_RF2保守结构域,推测其属于RF2类bZIP转录因子;对IbbZIP22蛋白进行亚细胞定位预测,推测该蛋白位于细胞核中;系统发育分析表明,IbbZIP22蛋白与三裂叶薯中的ItRF2a-like蛋白亲缘关系最近。采用实时定量PCR技术对济薯25和徐紫薯8号不同部位的IbbZIP22基因表达模式进行分析,发现IbbZIP22基因均在两个品种块根中表达量最高,且济薯25块根中的表达量显著高于徐紫薯8号,推测该基因参与淀粉调控。利用同源克隆技术,从济薯25基因组DNA中克隆IbbZIP22基因的启动子序列,该序列中除包含CAAT-box、TATA-box等核心启动子元件外,还存在多个参与光响应、抗逆和激素应答等元件。本研究结果为探究甘薯抗逆和淀粉调控机制提供了理论依据。In this study, the IbbZIP22 gene was cloned from Jishu 25. The IbbZIP22 gene coding region was 1 368 bp in full length and encoded 455 amino acids. Sequence and structural analysis showed that the bZIP_plant_RF2 conserved domain was found in the protein encoded by this gene, which was speculated to be a bZIP transcription factor of RF2 class. The IbbZIP22 protein was predicted to be located in the nucleus by subcellular localization. Phylogenetic analysis showed that IbbZIP22 protein was most closely related to ItRF2a-like protein in Ipomoea triloba. The expression pattern of IbbZIP22 gene in different parts of Jishu 25 and Xuzishu 8 was analyzed by real-time PCR. It was found that the expression level of IbbZIP22 gene in tuber root was the highest in both varieties, and the expression level in tuber root of Jishu 25 was significantly higher than that of Xuzishu 8, speculating that this gene was involved in starch regulation. The promoter sequence of IbbZIP22 gene was cloned from Jishu 25 genomic DNA by homologous cloning technique. In addition to the core promoter elements such as CAAT-box and TATA-box, the sequence also contained several elements involved in light response, stress resistance and hormone response. These results could provide a theoretical basis for exploring the mechanisms of resistance to stress and starch regulation in sweet potato.

关 键 词：甘薯 IbbZIP22基因 克隆 表达分析 启动子 淀粉 

分 类 号：S531[农业科学—作物学] Q785[生物学—分子生物学]