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Generation of Transgenic Maize by Two Germinating Seed Transformation Methods

文献类型： 外文期刊

作者： Liang, Xue-lian 1 ; Du, Jian-zhong 2 ; Hao, Yao-shan 2 ; Cui, Gui-mei 2 ; Wang, Yi-xue 2 ; Wang, Xiao-qing 2 ; Zhang, 1 ;

作者机构： 1.Zhongkai Univ Agr & Engn, Coll Life Sci, Guangzhou 510225, Guangdong, Peoples R China

2.Shanxi Acad Agr Sci, Minist Agr, Key Lab Crop Gene Resources & Germplasm Enhanceme, Biotechnol Res Ctr, Taiyuan 030031, Peoples R China

3.Shanxi Acad Agr Sci, Minist Agr, Key Lab Crop Gene Resources & Germplasm Enhanceme, Biotechnol Res Ctr, Taiyuan 030

关键词： Maize;Genetic transformation;Germinating seeds;Scarify;Puncture;Bar gene

期刊名称：INTERNATIONAL JOURNAL OF AGRICULTURE AND BIOLOGY （ 影响因子：0.822； 五年影响因子：0.906 ）

ISSN： 1560-8530

年卷期： 2016 年 18 卷 1 期

页码：

收录情况： SCI

摘要： Two germinating seed transformation methods i.e., scarifying and puncturing methods, were used to transfer the bar gene into an inbred maize line. We generated transgenic glufosinate-tolerant maize plants, and conducted a comparison of the two transformation methods. We used the plasmid pCAMBAR.CHI.11 as the donor and germinating maize seeds as the recipients. The germinating seeds were subjected to puncture or scarifying treatment after being soaked in water for 12 h. The injured seeds were co-cultivated with Agrobacterium for 24 h and then sown in the experimental plots. Putative transgenic maize plants were selected by basta tolerance screening and PCR detection, and were further confirmed by Southern blotting. Following glufosinate tolerance assay and molecular detection of transgenic lines of the transgenic seedling (T-0), first generation of transgenic seedling (T-1) and second generation of transgenic seedling (T-2), we have confirmed that the bar gene was stably inherited and expressed. The PCR amplification result for T-2 transgenic plants showed that the genetic segregation ratio of the bar gene followed the 3:1 ratio of a single dominant Mendelian factor. The study proved that both the germinating seed treatment approaches are rapid and simple plant transformation methods. In particular, the application of the puncture method can expand the tissue culture free transformation to dicotyledonous plants. (C) 2016 Friends Science Publishers

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