Development of lepidopteran pest-resistant transgenic japonica rice harboring a synthetic cry2A* gene

doi:10.1016/S2095-3119(14)60897-2

Journal of Integrative Agriculture ›› 2015, Vol. 14 ›› Issue (3): 423-429.DOI: 10.1016/S2095-3119(14)60897-2

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

收稿日期:2014-08-24 出版日期:2015-03-01 发布日期:2015-03-12
通讯作者: LIN Xiu-feng, Tel/Fax: +86-431-87063078,E-mail: linxiufeng8581@163.com
作者简介:JIN Yong-mei, E-mail: ymjin0303@163.com;* These authors contributed equally to this study.
基金资助:
This research was funded by the National Major Project for Transgenic Organism Breeding, China (201408001- 001-009).

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

Received:2014-08-24 Online:2015-03-01 Published:2015-03-12
Contact: LIN Xiu-feng, Tel/Fax: +86-431-87063078,E-mail: linxiufeng8581@163.com
About author:JIN Yong-mei, E-mail: ymjin0303@163.com;* These authors contributed equally to this study.
Supported by:
This research was funded by the National Major Project for Transgenic Organism Breeding, China (201408001- 001-009).

摘要/Abstract

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.

关键词: insect resistance , Bt gene , cry2A* transgenic rice , japonica rice variety , striped stem borer

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.

Key words: insect resistance , Bt gene , cry2A* transgenic rice , japonica rice variety , striped stem borer

JIN Yong-mei, MA Rui, YU Zhi-jing, WANG Ling, JIANG Wen-zhu, LIN Xiu-feng. Development of lepidopteran pest-resistant transgenic japonica rice harboring a synthetic cry2A* gene[J]. Journal of Integrative Agriculture, 2015, 14(3): 423-429.

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Development of lepidopteran pest-resistant transgenic japonica rice harboring a synthetic cry2A* gene

Development of lepidopteran pest-resistant transgenic japonica rice harboring a synthetic cry2A* gene

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