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Expression of Foreign Genes Demonstrates the Effectiveness of Pollen-Mediated Transformation in Zea mays

文献类型： 外文期刊

作者： Yang, Liyan 1 ; Cui, Guimei 1 ; Wang, Yixue 1 ; Hao, Yaoshan 1 ; Du, Jianzhong 1 ; Zhang, Hongmei 3 ; Wang, Changbiao; 1 ;

作者机构： 1.Shanxi Acad Agr Sci, Biotechnol Res Ctr, Taiyuan, Peoples R China

2.Shanxi Normal Univ, Coll Life Sci, Linfen, Peoples R China

3.Shanxi Acad Agr Sci, Maize Res Inst, Taiyuan, Peoples R China

4.Univ New England, Sch Environm & Rural Sci, Armidale, NSW, Australia

5.Minist Agr, Key Lab Crop Gene Resources & Germplasm Enhanceme, Taiyuan, Peoples R China

关键词： pollen-mediated transformation;sonication;eGfp;Cry1Ac;Zea mays

期刊名称：FRONTIERS IN PLANT SCIENCE （ 影响因子：5.753； 五年影响因子：6.612 ）

ISSN： 1664-462X

年卷期： 2017 年 8 卷

页码：

收录情况： SCI

摘要： Plant genetic transformation has arguably been the core of plant improvement in recent decades. Efforts have been made to develop in planta transformation systems due to the limitations present in the tissue-culture-based methods. Herein, we report an improved in planta transformation system, and provide the evidence of reporter gene expression in pollen tube, embryos and stable transgenicity of the plants following pollen-mediated plant transformation with optimized sonication treatment of pollen. The results showed that the aeration at 4 degrees C treatment of pollen grains in sucrose prior to sonication significantly improved the pollen viability leading to improved kernel set and transformation efficiency. Scanning electron microscopy observation revealed that the removal of operculum covering pollen pore by ultrasonication might be one of the reasons for the pollen grains to become competent for transformation. Evidences have shown that the eGfp gene was expressed in the pollen tube and embryos, and the Cry1Ac gene was detected in the subsequent T-1 and T-2 progenies, suggesting the successful transfer of the foreign genes to the recipient plants. The Southern blot analysis of Cry1Ac gene in T-2 progenies and PCR-identified Apr gene segregation in T-2 seedlings confirmed the stable inheritance of the transgene. The outcome illustrated that the pollen-mediated genetic transformation system can be widely applied in the plant improvement programs with apparent advantages over tissue-culture-based transformation methods.

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