油菜Bna-miR1140基因的分离与转化及其过表达分析  

Cloning and Transformation of Bna-miR1140 Gene and Its over Expression Analysis in Brassica napus

作　　者：董云[1] 王毅[1] 靳丰蔚[1] 吴旺泽 方彦[3] 徐妙云[4] 王磊[4] DONG Yun;WANG Yi;JIN Fengwei;WU Wangze;FANG Yan;XU Miaoyun;WANG Lei(Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou, Gansu 730070, China;College of Life Sciences , Northwest Normal University, Lanzhou, Gansu 730070;Gansu Provincial Key Lab of Aridland,Lanzhou, Gansu 730070, China;Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China)

机构地区：[1]甘肃省农业科学院作物研究所,兰州730070 [2]西北师范大学生命科学学院,兰州730070 [3]甘肃农业大学干旱生境作物学重点实验室,兰州730070 [4]中国农业科学院生物技术研究所,北京100081

出　　处：《西北植物学报》2018年第4期589-597,共9页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家科技重大专项(2018ZX08020001-008);国家自然科学基金(31260334);甘肃省农业科学院中青年基金(2015GAAS34)

摘　　要：为了探究油菜miR1140基因的功能,该研究利用PCR技术从油菜栽培种westar中克隆了miR1140前体基因,并与pPZP212载体连接,构建了miR1140前体基因的过表达载体,通过农杆菌介导法将其转化到甘蓝型油菜westar品种中。(1)PCR鉴定表明,共获得14株过表达Bna-miR1140基因油菜阳性株。(2)表型观察显示,阳性株的叶片形态、花器结构没有变异,株高、主花序长度、主花序结角数和千粒重均与对照相当,但其中有5株过表达Bna-miR1140基因油菜阳性株(T0代)出现双主序株型,且分枝部位较对照低,分枝数较对照明显增多,使全株有效角果数增多,单株生产力较对照提高26%,而其他9株阳性株均与野生型油菜表型一致。(3)T1代田间表型分析表明,5株过表达35S∷Bna-miR1140(T0代)的T1代转化株系全生育期较对照极显著延长9~14d,且5株T0代中有4株的株型变异符合3∶1的遗传分离规律。研究推测,油菜Bna-miR1140基因可能参与了调控油菜的分枝发育。In order to explore the function of the rape miR1140, we cloned the pre-miR1140 gene from rape cultivated variety westar by PCR approach, and successfully constructed the plant expression vector of pre-miR1140 with vector pPZP212, and transformed into variety westar by agrobacterium tumefaciens mediated approach. （1） PCR identification indicated that 14 strains positive transgenic plants of Bna-miR 1140 gene over expressed have been obtained; （2） Phenotypic observation showed that leaf morphology and flower tube of positive strains do no t mutate, plant height, length of main inflorescence, the number of principal inflorescences and the 1 000-grain weight are all equivalent to the control. But there are 5 positive strains with Bna-miR 1140 transformed （T 0） expressed double main sequence phenotype, and the branch height lower than control, branching number increased significantly than control, make the whole effective pod number increase, single productivity increased by 26% than that of control, and other 9 strains positive strains were consistent with that of wild type rape phenotype. （3） T 1 field phenotype analysis showed that the total growth period of T 1 transmutation plant of 5 strains with 35S∷ Bna-miR 1140 （T 0 generation） overexpressed was significantly increased by 9-14 d compared with the control group, and, of which there are four strains, their plant type variation genetic followed 3∶1 Mendel segregation law. The study speculated that Bna-miR 1140 may participate in the regulation of branching and development of oilseed rape.

关 键 词：油菜 分枝 miR1140 基因 过表达 

分 类 号：Q785[生物学—分子生物学] Q786