拟南芥Pri-miR156a基因对烟草开花时间的影响  

The Influences of Arabidopsis Pri-miR156a on Flowering Time of Nicotiana tabacum

作　　者：韩瑶瑶 马燕勤 李典珍 徐子勤[1] 

机构地区：[1]西部资源生物与现代生物技术教育部重点实验室,陕西省生物技术重点实验室,西北大学,生命科学学院,西安710069

出　　处：《基因组学与应用生物学》2015年第2期350-356,共7页Genomics and Applied Biology

基　　金：西北大学生命科学学院生物技术重点实验室;国家自然科学基金(30870194和J1210063);陕西省重点实验室科研计划(12JS103和2010JS090);西北大学研究生创新计划(YZZ13068);西部资源生物与现代生物技术教育部重点实验室开放研究基金共同资助

摘　　要：本研究利用转基因烟草分析了mi R156对SPL3基因的保守性调节作用。首先通过数据库搜索和RT-PCR方法克隆了普通烟草Ntab SPL3基因的编码序列,并进行了测序验证。同时从拟南芥Col-0生态型基因组DNA分离了At Pri-mi R156a基因序列,构建产生植物表达载体,采用农杆菌转化技术制备了转基因烟草植株。RT-PCR分析发现,过量表达Atmi R156a可以明显下调烟草Ntab SPL3基因。表型观察结果显示,At Pri-mi R156a转基因烟草个体矮小,开花时间明显延迟,叶片数量及生物量积累增多,说明组成性表达Atmi R156a能够延长普通烟草的营养生长时间,推迟开花转变过程。本研究为培育适合不同生长环境的烟草新品种提供了一条新的途径,对烟草改良具有重要意义。The conservative regulation function of mi R156 to SPL3 was investigated in the present work. The coding sequence of tobacco NtabSPL3 was cloned by RT-PCR based on the results of homologous search to databases, and was confirmed by sequencing. Simultaneously, the genomic DNA sequence of At Pri-mi R156 a isolated from Arabidopsis ecotype Col-0 was used to construct a plant expression vector and transgenic tobacco plants were prepared by Agrobacterium-mediated transformation method. RT-PCR analysis revealed that NtabSPL3 could be downregulated obviously by overexpression of Atmi R156 a. Compared to wide-type tobacco plants, the transgenic tobacco plants harbouring the Arabidopsis Pri-mi R156 a showed a number of changed phenotypic characteristics, including dwarfism in size, delay in floral transition and increases in leaf number and biomass accumulation, indicating constitutive expression of Atmi R156 a could extend the time of vegetative growth and postpone the floral transition in tobacco. This work provides a novel approach for breeding of new tobacco cultivars adaptable to different circumstances, and is valuable in tobacco improvement.

关 键 词：烟草 秦烟95 MicroRNA156 开花时间 NtabSPL3 

分 类 号：S572[农业科学—烟草工业]