Development of Transgenic Maize Plants Tolerant to Glyphosate via Pollen-mediated Transformation and Their Glyphosate Tolerance

doi:10.13560/j.cnki.biotech.bull.1985.2016.11.018

Abstract

Abstract: In order to develop transgenic maize plants tolerant to glyphosate and further study their herbicide tolerance and the genetic characteristics of exogenous gene in transgenic plants,plasmid pBI101-aroA-M12 harboring aroA-M12 gene was transformed into maize inbred Chang ‘7-2' plants by pollen-mediated transformation method. Fifty-four transgenic plants were obtained by PCR detection and Southern blot analysis. The detection results of molecular analysis and bioassay in the field showed that target aroA-M12gene was integrated into the genome of transgenic plants and could stably inherit to next generation. Moreover,the aroA-M12 gene conferred the glyphosate-tolerance to transgenic plants. The result of genetic rule of target gene in transgenic plants suggested that the target gene was segregated according to the ratio of 3∶1 in T₂ generation transgenic plants（lines）and it was consistent with the Mendel law of single-factor heredity. The results of ELISA analysis and protein concentration determination of transgenic plants confirmed that the target gene was expressed in transgenic plants,and the protein amount of gene expression were 40.5-112.6 ng/g leaf fresh weight,there was a significant correlation between protein amount of expression of each transgenic plant and corresponding glyphosate-tolerance of that plant,correlation coefficients r = 0.942 3（P<0.01）. Ultimately,4 pure lines of transgene with high glyphosate-tolerance were obtained from this experiment.

Key words: maize, aroA-M12 gene, glyphosate-tolerance, pollen-mediated transformation, genetic analysis

YANG Hui-zhen, REN Zhi-qiang, XIAO Jian-hong, BU Hua-hu, LIU Hui-min. Development of Transgenic Maize Plants Tolerant to Glyphosate via Pollen-mediated Transformation and Their Glyphosate Tolerance[J]. Biotechnology Bulletin, 2016, 32(11): 152-161.

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