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Development of lepidopteran pest-resistant transgenic japonica rice harboring a synthetic cry2A* gene |
JIN Yong-mei, MA Rui, YU Zhi-jing, WANG Ling, JIANG Wen-zhu, LIN Xiu-feng |
1、Agro-Biotechnology Research Institute, Jilin Academy of Agricultural Sciences, Changchun 130124, P.R.China
2、School of Life Science, Jilin Agricultural University, Changchun 130118, P.R.China
3、College of Plant Science, Jilin University, Changchun 130062, P.R.China |
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摘要 A synthetic cry2A* gene encoding Bacillus thuringiensis (Bt) δ-endotoxin that resistance to lepidopteran pest was transformed into japonica rice variety Jijing 88, which is the most widely cultivated variety in Jilin Province, Northeast China, by Agrobacterium-mediated transformation. A total of 106 independent transformants overexpressing cry2A* gene driven by ubiquitin (Ubi) promoter was produced. Three single-copy homozygous transgenic lines were finally selected based on the results of PCR analysis, segregation ratio of Basta resistance, and Southern hybridization analyses. RT-PCR and enzyme linked immune sorbent assay (ELISA) revealed that cry2A* transcripts and protein were highly expressed in these lines. The high level of Cry2A* protein expression resulted in high resistance to rice striped stem borer as evidenced by insect feeding bioassays. Our results demonstrate that cry2A* transgenic japonica rice confers resistance to the rice striped stem borer in the laboratory conditions.
Abstract A synthetic cry2A* gene encoding Bacillus thuringiensis (Bt) δ-endotoxin that resistance to lepidopteran pest was transformed into japonica rice variety Jijing 88, which is the most widely cultivated variety in Jilin Province, Northeast China, by Agrobacterium-mediated transformation. A total of 106 independent transformants overexpressing cry2A* gene driven by ubiquitin (Ubi) promoter was produced. Three single-copy homozygous transgenic lines were finally selected based on the results of PCR analysis, segregation ratio of Basta resistance, and Southern hybridization analyses. RT-PCR and enzyme linked immune sorbent assay (ELISA) revealed that cry2A* transcripts and protein were highly expressed in these lines. The high level of Cry2A* protein expression resulted in high resistance to rice striped stem borer as evidenced by insect feeding bioassays. Our results demonstrate that cry2A* transgenic japonica rice confers resistance to the rice striped stem borer in the laboratory conditions.
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Received: 24 August 2014
Accepted:
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Fund: This research was funded by the National Major Project for Transgenic Organism Breeding, China (201408001- 001-009). |
Corresponding Authors:
LIN Xiu-feng, Tel/Fax: +86-431-87063078,E-mail: linxiufeng8581@163.com
E-mail: linxiufeng8581@163.com
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About author: JIN Yong-mei, E-mail: ymjin0303@163.com;* These authors contributed equally to this study. |
Cite this article:
JIN Yong-mei, MA Rui, YU Zhi-jing, WANG Ling, JIANG Wen-zhu, LIN Xiu-feng.
2015.
Development of lepidopteran pest-resistant transgenic japonica rice harboring a synthetic cry2A* gene. Journal of Integrative Agriculture, 14(3): 423-429.
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