[1]陈李淼,田星星,单志慧,等.利用农杆菌介导法转化大豆子叶节的影响因素研究[J].大豆科学,2012,31(01):17-23.[doi:10.3969/j.issn.1000-9841.2012.01.005]
　CHEN Li-miao,TIAN Xing-xing,SHAN Zhi-hui,et al.Optimization of the Factors Affecting Genetic Transformation of Soybean Cotyledonary Node Mediated by Agrobacterium tumefacines[J].Soybean Science,2012,31(01):17-23.[doi:10.3969/j.issn.1000-9841.2012.01.005]
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利用农杆菌介导法转化大豆子叶节的影响因素研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第31卷 期数: 2012年01期 页码: 17-23 栏目: 出版日期: 2012-02-25
Title:
Optimization of the Factors Affecting Genetic Transformation of Soybean Cotyledonary Node Mediated by Agrobacterium tumefacines
文章编号:
1000-9841（2012）01-0017-07
作者:
陈李淼1; 2; 3; 田星星1; 2; 单志慧1; 2; 沙爱华1; 2; 李文滨3; 周新安1; 2
1.中国农业科学院 油料作物研究所，湖北 武汉430062;
2.农业部油料作物生物学重点开放实验室，湖北 武汉 430062;
3.东北农业大学 国家教育部大豆生物学重点实验室，黑龙江 哈尔滨150030
Author(s):
CHEN Li-miao1; 2; 3; TIAN Xing-xing1; 2; SHAN Zhi-hui1; 2; SHA Ai-hua1; 2; LI Wen-bin3; ZHOU Xin-an1; 2
1.Oil Crops Research Institute of Chinese Academy of Agricultural Sciences(CAAS),Wuhan 430062,Hubei；
2.Key Laboratory of Oil Crop Biology,Ministry of Agriculture,Wuhan 430062,Hubei；
3.Key Laboratory of Soybean Biology,Ministry of Education,Soybean Research Institute,Northeast Agriculture University,Harbin 150030,Heilongjiang,China
关键词:
子叶节; 胚尖; 基因型; 侵染浓度; 侵染时间; 乙酰丁香酮; 超声波
Keywords:
Cotyledonary node; Embryonic tip; Genotype; A.tumefacines concentration; Infection time; Acetosyringone; Sonication
分类号:
S565.1
DOI:
10.3969/j.issn.1000-9841.2012.01.005
文献标志码:
A
摘要:
以3个大豆品种为材料，采用GUS瞬时表达的方法，对适合于大豆子叶节和胚尖转化的基因型进行筛选，结果表明：适合大豆子叶节转化的品种为东农50，适合胚尖转化的品种为黑农41。在子叶节转化系统中，摸索了农杆菌侵染浓度与侵染时间的最佳配比组合、乙酰丁香酮浓度和超声波辅助处理对大豆转化效率的影响。优化后的条件为：农杆菌侵染的浓度OD600=0.6，侵染时间30~40 min，乙酰丁香酮浓度200 μmol·L-1，超声波辅助转化时间5 s。在上述条件下，成功获得了转基因植株，转化效率为2.3%。
Abstract:
Three soybean genotypes were used for optimal genotype selection in transformation mediated by Agrobacterium tumefacines.The influences of A.tumefacinesconcentration,infection time,acetosyringone(AS) concentration and sonication treatment were studied in this research.As a result,the appropriate genotype for cotyledonary node transformation was Dongnong 50 while Heinong 41 was fit for embryo tip transformation.In the cotyledonary node transform system,the optimal infection combination was A.tumefacines concentration at OD600=0.6 with 30-40 min infection time.The perfect AS concentration was 200 μmol·L-1.The appropriate sonicate treatment was 5 s under 100 W.The study optimized soybean cotyledonary-node Agrobacterium-mediated transformation system and transformation frequency was 2.3%.

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备注/Memo

基金项目：转基因生物新品种培育重大专项（2008ZX08004-005）；中国农业科学院公益性院所科研基金（1610172011005）。

第一作者简介：陈李淼（1983-），女，博士，研究方向为大豆生物技术，大豆分子克隆与遗传转化。E-mail:ccllmm_008@yahoo.com.cn。
通讯作者：周新安 (1963-), 男, 研究员, 博士，主要从事大豆遗传育种研究。E-mail:xazhou@public.wh.hb.cn。

更新日期/Last Update: 2014-08-14