Chinese Bulletin of Botany ›› 2013, Vol. 48 ›› Issue (6): 616-622.DOI: 10.3724/SP.J.1259.2013.00616

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Cytological Study of Pollen-mediated Plant Transformation Method Based on GFP Observation

Na Li1, Yi Sun2,3, Liyan Yang1*   

  1. 1School of Life Sciences, Shanxi Normal University, Linfen 041004, China;

    2Biotechnology Research Centre, Shanxi Academy of Agricultural Sciences, Taiyuan 030031, China;

    3Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau, Ministry of Agriculture, Taiyuan 030031, China
  • Received:2013-01-30 Revised:2013-04-21 Online:2013-11-01 Published:2013-12-03
  • Contact: Liyan Yang

Abstract: Pollen-mediated gene transformation does not require tissue culture and is easy to apply. In the present study, we investigate the cytological basis for the transformation approach. Fresh pollen was taken from maize plants (Zea mays cv ‘Zhengdan 958’) and treated under ultrasonication mixed with plasmid DNA harboring green fluorescence protein (GFP) gene. Treated pollen was artificially pollinated onto silks of maize ears. The expression of GFP gene in pollen grains, pollen tubes and embryo were observed under a fluorescence microscope. Pollen grains had a strong green autoflorescent background and so were not suitable for observing the expression of exotic GFP gene. Treated pollen tubes produced strong green florescence as compared with the control. The expression of GFP gene in embryo could be used to confirm the expression of exotic genes. Our study provides cytological evidence for pollen-mediated transformation.

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