[1]周思军李希臣刘昭军刘丽艳杨庆凯.通过农杆菌介导法将Bt(cryIA)基因导入大豆[J].大豆科学,2001,20(03):157-162.[doi:10.11861/j.issn.1000-9841.2001.03.0157]
　Zhou SijunLi XichenLiu ZhaojunLiu LiyanYang Qingkai.INTRODUCTION OF Bt GENE(cryIA)INTO SOYBEANBY Agrobacterium -MEDIATED TRANSFORMTION[J].Soybean Science,2001,20(03):157-162.[doi:10.11861/j.issn.1000-9841.2001.03.0157]
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通过农杆菌介导法将Bt(cryIA)基因导入大豆

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第20卷 期数: 2001年03期 页码: 157-162 栏目: 出版日期: 2001-08-25
Title:
INTRODUCTION OF Bt GENE(cryIA)INTO SOYBEANBY Agrobacterium -MEDIATED TRANSFORMTION
文章编号:
1000 -9841(2001)03 -0157 -06
作者:
周思军1李希臣1刘昭军1刘丽艳1杨庆凯2
1.黑龙江省农业科学院生物技术研究中心, 哈尔滨150086 ;
2.东北农业大学大豆研究所, 150030
Author(s):
Zhou Sijun1Li Xichen1Liu Zhaojun1Liu Liyan1Yang Qingkai2
1.Biotechnology Research Center , Heilongjiang Academy of Agricultural Sciences , Harbin 150086 ;
2.Insti tute of Soybean , Northeast Agricultural University , Harbin 150030
关键词:
大豆; 转基因植株; Bt 基因; 转化
Keywords:
Soybean ; Transgenic plant ; Bt gene ; Transformation
分类号:
S565.035.3
DOI:
10.11861/j.issn.1000-9841.2001.03.0157
文献标志码:
A
摘要:
采用农杆菌介导的大豆子叶节转化系统成功地将Bt 基因(cry IA)导入大豆。从发芽5 -7天的大豆无菌苗切取子叶节外植体, 经农杆菌感染和共培养后, 在选择培养基上4 周左右出现抗性不定芽。将不定芽转移到芽伸长培养基上, 4 -6 周后再生苗长至2 .5 -3cm 高。再将再生苗切下转入生根培养基, 2 周左右生根。生根后的再生植株经逐步锻炼移入盆中, 所有植株均能正常开花结荚。在移栽成活的8 株T0 植株中有7 株PCR 检测呈阳性反应;在7 个T1 株系中有4 个株系存在PCR 阳性植株。取4 个稳定遗传的T1 代株系内的阳性植株的叶片提取DNA , 用地高辛标记的Bt 基因探针进行Southern 杂交分析, 结果4 个株系均呈现阳性, 证明Bt 基因已整合到受体大豆的基因组内并能传递给后代。
Abstract:
Baci llus thuringiensis cry IA gene w as int roduced into soybean successfully w ith Agrobacterium -cotyledo nary node t ransformation system .Cotyledonary node ex plants were prepared f rom 5 -7 day oldseedlings , infected and co -cultivated with Agrobacterium tumefaciens .Resistant adventitious buds emerged after4 weeks on selective medium .The buds w ere t ransferred onto elongation medium and regenerated shoot sg rew to 2 .5 -3 cm high af ter 4 -6 w eeks .The shoo ts were cut off and t ransferred onto root ing medium .Af terabout 3 weeks , the rooted plantlets w ere hardened of f and t ransferred into pots .All the plant s could f low er andset seeds normally .Among the 8 t ransplanted survival T0 plant s, seven gave posi tive PCR reaction ;and 4 of the7 T1 plant lines had PCR posi tive plant s .DNA w as ex tracted f rom the leaves of positive plants in the 4 T1 plantlines , and southern blo t w as conducted with probe of Bt gene .All the 4 lines are positive , indicating that the Btgene has integrated into the receiver’ s genome and can be passed to progenies .

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备注/Memo

作者简介:周思军,(1957 -), 男, 博士, 研究员, 从事植物转基因研究。

更新日期/Last Update: 2016-12-29