A simple and efficient method for obtaining transgenic soybean callus tissues
A simple and efficient method for obtaining transgenic soybean callus tissues
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一种简单高效获得转基因大豆愈伤组织的方法
DOI：
10.1007/s11738-013-1248-3
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发表时间：
2013-03
影响因子：
2.6
通讯作者：
Wang, Ning Ning
中科院分区：
文献类型：
--
作者：
Chang, Dasong;Wang, Lu;Wang, Dan;Wang, Ning Ning
本研究建立了一种简便、高效的获得大豆[Glycine max（L.）梅尔。]在农杆菌介导法转化的基础上，大豆下胚轴切段用作起始材料。研究了大豆基因型、农杆菌浓度、接种时间、共培养时间、共培养培养基中抗氧化剂的添加等因素对转化效率的影响。将外植体培养在含有0.5 mg L-16-苄氨基嘌呤和2.0 mg L-1，2，4-二氯苯氧基乙酸的愈伤组织诱导培养基上，用于愈伤组织诱导。通过优化的诱导方案，获得了CaMV 35 S：：GUS和CaMV 35 S：：GFP转基因愈伤组织。经GUS检测，平均转化效率达87.7%.从接种农杆菌到获得转基因大豆愈伤组织需要3周左右的时间。为了验证该方法在基因功能研究中的有效性，获得了转GVG：：GmSAR K基因大豆愈伤组织，并对其衰老相关表型进行了分析。据我们所知，这是第一个报告使用下胚轴片段作为起始材料，获得转基因愈伤组织，该系统提供了一种方法，高通量筛选功能基因的目的转化大豆愈伤组织。
In the present study, a simple and efficient method for obtaining transgenic callus tissues of soybean [Glycine max(L.) Merr.] was developed based onAgrobacterium-mediated transformation. Hypocotyl segments of soybean were used as the starting material. Several factors such as soybean genotype,Agrobacteriumconcentration, inoculation time, co-cultivation period and addition of antioxidants in co-cultivation medium affecting the transformation efficiency were examined. The explants were cultured on callus induction medium containing 0.5 mg L−16-benzylaminopurine and 2.0 mg L−1, 2,4-Dichlorophenoxyacetic acid for callus induction. Callus tissues were induced at both the acropetal and basipetal ends.CaMV35S::GUSandCaMV35S::GFPtransgenic callus tissues were obtained using the optimized protocol. The average transformation efficiency reached up to 87.7 % based on GUS detection. From inoculation withAgrobacteriumto obtaining transgenic soybean callus will take about 3 weeks. In order to validate this method for gene function investigation,GVG::GmSARKtransgenic soybean callus tissues were obtained and their senescence-associated phenotypes were assessed. To our knowledge, this is the first report using hypocotyl segments as starting materials to obtain transgenic callus, and this system provides a method for high-throughput screening of functional genes of interest in transformed soybean callus.
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影响因子：
4.2
作者：
J. Staden;F. E. Drewes
通讯作者：
J. Staden;F. E. Drewes
影响因子：
2.4
作者：
N. Jiang;Eun-Hee Jeon;J. Pak;T. Ha;I. Baek;W. Jung;Jai‐Heon Lee;D. Kim;Hong-Kyu Choi;
通讯作者：
N. Jiang;Eun-Hee Jeon;J. Pak;T. Ha;I. Baek;W. Jung;Jai‐Heon Lee;D. Kim;Hong-Kyu Choi;
影响因子：
5.4
作者：
B. Reisch;S. H. Duke;E. Bingham
通讯作者：
B. Reisch;S. H. Duke;E. Bingham
DOI：
10.1002/j.1460-2075.1987.tb02730.x
发表时间：
1987-12
期刊：
The EMBO Journal
影响因子：
--
作者：
R. Jefferson;T. Kavanagh;M. Bevan
通讯作者：
R. Jefferson;T. Kavanagh;M. Bevan
影响因子：
0.6
作者：
R. Chand;D. Sen;K. Prasad;A. Singh;B. Bashyal;L. Prasad;A. Joshi
通讯作者：
R. Chand;D. Sen;K. Prasad;A. Singh;B. Bashyal;L. Prasad;A. Joshi