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Characterization of transformed soybean strains expressing GbNPR1 and HrpZpsg12 genes for disease resistance

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Abstract

To improve soybean broad-spectrum disease resistance, a bivalent plant expression vector carrying the GbNPR1 and HrpZpsg12 genes with the Bar gene as a selectable marker was constructed and transformed into soybean cultivar ‘Jinong 28’ using an Agrobacterium-mediated method. The results of PCR, Southern blot and SDS-PAGE analyses indicated that the two disease resistance genes, GbNPR1 and HrpZpsg12, and the selectable marker gene, Bar, were successfully integrated into the genome of soybean Jinong 28 and the proteins encoded by the inserted genes were expressed in the transgenic plants. Real-time quantitative PCR results at T2 to T4 revealed that relative expression levels of two disease resistance genes, GbNPR1 and HrpZpsg12 were similar, both highest in leaves, followed by roots, and much lower in stem and matured seed. The relative expression levels of these genes at T3 and T4 generations were similar and much higher than that at T2 generation, implying that transgenic strains were stably inherited to T4 generation. The disease resistance of transgenic strains was significantly increased to moderately resistant to both Phytophthora sojae and Cercospora sojina with artificial inoculation methods. Field evaluation results with artificial infection demonstrated that the T3 and T4 transgenic strains had significantly improved the resistance to both P. sojae and C. sojina when compared with the transformation donor.

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Acknowledgments

We thank the Jilin Academy of Agricultural Sciences for providing the plasmid containing the GbNPR1 gene and the Department of Plant Pathology of Jilin Agricultural University for providing the HrpZpsg12 gene. This research was funded by Grants from the National Science and Technology Major Project (2011ZX08004-004).

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Authors and Affiliations

  1. Center for Plant Biotechnology, Jilin Agricultural University, Changchun, Jilin, People’s Republic of China

    Yang Song, Xueming Zhang, Piwu Wang, Huanyu Dong, Jian Ma, Yongping Fu & Zhuo Zhang

  2. Seed Management Division of Changchun, Changchun, Jilin, People’s Republic of China

    Zhenku Liu

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  1. Yang Song
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  2. Xueming Zhang
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  3. Piwu Wang
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  4. Zhenku Liu
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  7. Yongping Fu
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  8. Zhuo Zhang
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Correspondence to Piwu Wang.

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Song, Y., Zhang, X., Wang, P. et al. Characterization of transformed soybean strains expressing GbNPR1 and HrpZpsg12 genes for disease resistance. Euphytica 211, 369–377 (2016). https://doi.org/10.1007/s10681-016-1749-4

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