甘蓝转录因子BoLH27的克隆与转基因甘蓝的表型分析  

Cloning of BoLH27 Gene from Cabbage and Phenotype Analysis of Transgenic Cabbage

作　　者：梁云飞 张林成 蒲全明 雷镇泽 施松梅 姜宇鹏 任雪松 高启国 LIANG Yun-Fei;ZHANG Lin-Cheng;PU Quan-Ming;LEI Zhen-Ze;SHI Song-Mei;JIANG Yu-Peng;REN Xue-Song;GAO Qi-Guo(College of Horticulture and Landscape Architecture,Southwest University/Key Laboratory of Horticulture Science for Southern Mountainous Regions,Ministry of Education,Chongqing 400716,China;Institute of Vegetables,Nanchong Academy of Agricultural Sciences,Nanchong 637000,Sichuan,China)

机构地区：[1]西南大学园艺园林学院/南方山地园艺学教育部重点实验室,重庆400716 [2]南充市农业科学院蔬菜研究所,四川南充637000

出　　处：《作物学报》2018年第3期397-404,共8页Acta Agronomica Sinica

基　　金：国家重点基础研究发展计划项目(973计划)(2012CB113900);国家自然科学基金项目(30900986)资助~~

摘　　要：bHLH转录因子对植物生长发育、形态调控有重要作用,为了研究BoLH27基因在甘蓝叶片发育及形态建成中的调控功能,本文以甘蓝品种519为材料,克隆出转录因子BoLH27基因。序列分析表明,该基因含有一个795 bp的开放阅读框,编码264个氨基酸,具有一个保守的典型bHLH结构域。以农杆菌介导的遗传转化方法将正义BoLH27基因导入甘蓝品种519中以增强表达,通过PCR筛选出9株T0代转基因单株,结合T2代单株的q RT-PCR分析表明,筛选的转基因植株内BoLH27基因的表达积累量明显高于野生型。试验基地隔离网内种植的转基因甘蓝表型明显,主要表现为莲座期茎叶间距拉长、叶柄伸长以及植株茎和叶柄显示紫色,植株叶片平展,无向上向内卷曲趋势,表明BoLH27基因可能对甘蓝叶片发育有重要的调控作用。bHLH transcription factor plays an important role in plant growth,development and morphological control.In order to explore BoLH27 gene regulatory function for leaf development and morphologic formation,the BoLH27 gene was cloned from cabbage(Brassica oleracea L.)variety 519.Sequence analysis indicated that the length of BoLH27 gene was 795 bp,which encoded 264 amino acids.The BoLH27 protein contained conservative structure domains of the bHLH family.The sense BoLH27 gene was transformed into cabbage variety 519 by Agrobacterium mediated method,PCR analysis exhibited that BoLH27 was genetically transformed into nine individual plants,qRT-PCR analysis revealed that BoLH27 had higher expression level in T2 transgenic cabbage than in wild type.The phenotype at rosette stage of transgenic cabbage grown in the test base was obvious,showing elongated the stems and leaves,elongated petiole,purple stems and petiole,and flat leaves without upward inward curling trend.It suggested that BoLH27 may play important roles in the control of leaves development.

关 键 词：甘蓝 BoLH27 基因克隆 转基因 表型 

分 类 号：S635[农业科学—蔬菜学]