甘蓝型油菜BnDGP1基因的克隆与表达分析  

Cloning and expression analysis of BnDGP1 gene in Brassica napus

作　　者：彭泽川 赵东方 洪波 李诗珺 周冰倩 李茜 官春云[1] 官梅[1] PENG Zechuan;ZHAO Dongfang;HONG Bo;LI Shijun;ZHOU Bingqian;LI Xi;GUAN Chunyun;GUAN Mei(College of Agronomy,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学农学院,长沙410128

出　　处：《植物生理学报》2023年第5期878-888,共11页Plant Physiology Journal

基　　金：国家现代农业产业技术体系(CARS-13);湖南省现代农业产业技术体系。

摘　　要：DGP1(DEEP GREEN PANICLE1)是植物的一种特异性蛋白,具有保守的TIGR01589结构域,在绿色组织中表达。为探究DGP1基因在甘蓝型油菜中的功能,本研究以甘蓝型油菜‘高油酸1号’为材料,克隆得到2个同源拷贝,长度为288 bp,编码95个氨基酸,分别命名BnDGP1-1和BnDGP1-2。生物信息学分析结果表明,BnDGP1-1、BnDGP1-2与拟南芥AT3G55240及水稻OsDGP1分子式相近,均为不稳定亲水性非跨膜蛋白,未找到信号肽结构,均属于非分泌蛋白;BnDGP1-1、BnDGP1-2的二级结构和三级结构相吻合。亚细胞定位结果表明,BnDGP1-1、BnDGP1-2均在细胞核中表达。实时荧光定量PCR分析表明,BnDGP1的5个同源拷贝除BnaA05G0066400ZS在花中相对表达最高,其他拷贝表达趋势没有明显差异,均为叶片>茎>角果皮>花的表达模式,而在根和花蕾中几乎不表达。本文构建了植物表达载体pCAMBIA1304-Bn-DGP1-1、pCAMBIA1304-BnDGP1-2以及CRISPR-cas9敲除载体。DGP1(DEEP GREEN PANICLE1)is a plant specific protein with a conserved TIGR01589 domain,which is expressed in green tissue.In order to explore the function of DGP1 gene in Brassica napus,this study used high oleic acid 710 as material,cloned two homologous copies,288 bp in length,encoding 95 amino acids,named BnDGP1-1 and BnDGP1-2 respectively.The results of bioinformatics analysis showed that BnDGP1-1 and BnDGP1-2 were similar to AT3G55240 of Arabidopsis thaliana and OsDGP1 of rice in molecular formula,and they were unstable hydrophilic non transmembrane proteins.No signal peptide structure was found,so they were non-secretory proteins.The secondary structure of BnDGP1-1 and BnDGP1-2 is consistent with the tertiary mechanism.The subcellular localization results showed that BnDGP1-1 and BnDGP1-2 were expressed in the nucleus.Real time fluorescence quantitative PCR analysis showed that the five homologous copies of BnDGP1 had the highest relative expression in the flower,except BnaA05G0066400ZS,the other copies had no significant difference in expression trend,and all showed the expression pattern of leaf>stem>hornfruit peel>flower,while almost no expression in the root and bud.Plant expression vectors pCAMBIA1304-BnDGP1-1,pCAMBIA1304-BnDGP1-2 and CRISPR-cas9 knockout vectors were constructed.The results provide some reference for BnDGP1 gene to regulate chloroplast development and photosynthesis in B.napus.

关 键 词：甘蓝型油菜 DGP1 生物信息学分析 载体构建 光合作用 

分 类 号：S565.4[农业科学—作物学]