title: Expression of Foreign Genes Demonstrates the Effectiveness of Pollen-Mediated Transformation inZea mays.
abstract: 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°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 theeGfpgene was expressed in the pollen tube and embryos, and theCry1Acgene was detected in the subsequent T1and T2progenies, suggesting the successful transfer of the foreign genes to the recipient plants. The Southern blot analysis ofCry1Acgene in T2progenies and PCR-identifiedAprgene segregation in T2seedlings 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.
authors.affiliation: Biotechnology Research Center, Shanxi Academy of Agricultural SciencesTaiyuan, China; College of Life Science, Shanxi Normal UniversityLinfen, China.
authors.forename: Liyan
authors.initials: L
authors.lastname: Yang
authors.affiliation: Biotechnology Research Center, Shanxi Academy of Agricultural Sciences Taiyuan, China.
authors.forename: Guimei
authors.initials: G
authors.lastname: Cui
authors.affiliation: Biotechnology Research Center, Shanxi Academy of Agricultural Sciences Taiyuan, China.
authors.forename: Yixue
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Biotechnology Research Center, Shanxi Academy of Agricultural Sciences Taiyuan, China.
authors.forename: Yaoshan
authors.initials: Y
authors.lastname: Hao
authors.affiliation: Biotechnology Research Center, Shanxi Academy of Agricultural Sciences Taiyuan, China.
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Du
authors.affiliation: Maize Research Institute, Shanxi Academy of Agricultural Sciences Taiyuan, China.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Biotechnology Research Center, Shanxi Academy of Agricultural Sciences Taiyuan, China.
authors.forename: Changbiao
authors.initials: C
authors.lastname: Wang
authors.affiliation: Biotechnology Research Center, Shanxi Academy of Agricultural Sciences Taiyuan, China.
authors.forename: Huanhuan
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Biotechnology Research Center, Shanxi Academy of Agricultural SciencesTaiyuan, China; School of Environmental and Rural Science, University of New England, ArmidaleNSW, Australia.
authors.forename: Shu-Biao
authors.initials: SB
authors.lastname: Wu
authors.affiliation: Biotechnology Research Center, Shanxi Academy of Agricultural SciencesTaiyuan, China; Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau, Ministry of AgricultureTaiyuan, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Sun
keywords: Cry1Ac; Zea mays; eGfp; pollen-mediated transformation; sonication