[1]郝荣华,邵 群,杨素欣,等.根癌农杆菌介导的大豆子叶节转化体系的优化[J].大豆科学,2012,31(02):167-172.[doi:10.3969/j.issn.1000-9841.2012.02.002]
　HAO Rong-hua,SHAO Qun,YANG Su-xin,et al.Optimization of Agrobacterium-mediated Soybean Transformation using the Cotyledonary Node[J].Soybean Science,2012,31(02):167-172.[doi:10.3969/j.issn.1000-9841.2012.02.002]
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根癌农杆菌介导的大豆子叶节转化体系的优化

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第31卷 期数: 2012年02期 页码: 167-172 栏目: 出版日期: 2012-04-25
Title:
Optimization of Agrobacterium-mediated Soybean Transformation using the Cotyledonary Node
文章编号:
000-9841（2012）02-0167-06
作者:
郝荣华1; 邵 群1; 杨素欣1; 李继存2; 冯献忠1; 赵彦修1
1.山东师范大学 生命科学学院，山东省高校系统生物学重点实验室，山东 济南 250014；
2.济宁市农业科学院 经济作物所，山东 济宁 272031
Author(s):
HAO Rong-hua1; SHAO Qun1; YANG Su-xin1; LI Ji-cun2; FENG Xian-zhong1; ZHAO Yan-xiu1
1.Key Laboratory of Systems Biology in Universities of Shandong,College of Life Science,Shandong Normal University,Jinan 250014;
2.Economic Plants Institute,Jining Academy of Agricultural Sciences,Jining 272031,Shandong,China
关键词:
大豆; 农杆菌; 子叶节; 转化
Keywords:
Soybean; Agrobacterium tumefaciens; Cotyledonary node; Transformation
分类号:
S565.1
DOI:
10.3969/j.issn.1000-9841.2012.02.002
文献标志码:
A
摘要:
以13种不同基因型的大豆为材料，研究了大豆外植体再生过程中不同大豆基因型对丛生芽数目的影响，选取优势基因型为受体材料，比较了在草丁膦和潮霉素筛选压力下外植体的再生情况，并确定其合适的筛选浓度，在对农杆菌介导的子叶节转化体系优化的基础上进行了大豆转化效率的研究。结果显示：在相同的芽诱导条件下，山宁14产生的丛生芽最多，更适合于大豆子叶节转化；与其它筛选剂相比，采用潮霉素的梯度筛选更有利于抗性苗的成活，其最适筛选浓度为8 mg·L-1；通过检测GUS基因的表达和分子鉴定证明外源的GUS基因已插入到转基因植株的基因组中，转化效率达到3.2%。
Abstract:
To optimize the system of Agrobacterium-mediated soybean\[Glycine max (L.) Merrill\]transformation,effects of 13 cultivars and 2 selection agents were studied during the process of shoot regeneration and transformation.The results showed that the cultivar ‘Shanning 14’ induced the highest number of multiple shoot among tested 13 cultivars,and hygromycin B was better than glufosinate during transformant selection of cultivar ‘Shanning 14’.The optimal transformant selection scheme was a gradient concentration of hygromycin B from 3 to 8 mg·L-1.The integrations of UidA(GUS)gene into the transformant genome were demonstrated by histological staining,PCR application and sequencing analysis.Transformation efficiency was 3.2% based on the number of transformed events.

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备注/Memo

基金项目：山东省自然科学杰出青年基金资助项目（JQ200909）；国家自然科学基金资助项目（31171571）。

第一作者简介：郝荣华（1983-），女，在读博士，研究方向为大豆转基因。E-mail:ronghua34@163.com。
通讯作者：赵彦修(1954-)，男，教授，从事植物耐盐细胞及分子生物学研究。E-mail:zhaoyx@sdnu.edu.cn。

更新日期/Last Update: 2014-08-15