Acquisition of Insect-Resistant Transgenic Maize Harboring a Truncated cry1Ah Gene via Agrobacterium-Mediated Transformation
文献类型: 外文期刊
第一作者: Huang Da-fang
作者: Huang Da-fang;Li Xiu-ying;Lang Zhi-hong;Zhu Li;Huang Da-fang;Zhang Jie;He Kang-lai
作者机构:
关键词: Bacillus thuringiensis;truncated cry1Ah gene;insectresistant maize;stable inheritance
期刊名称:JOURNAL OF INTEGRATIVE AGRICULTURE ( 影响因子:2.848; 五年影响因子:2.979 )
ISSN: 2095-3119
年卷期: 2014 年 13 卷 5 期
页码:
收录情况: SCI
摘要: A novel insecticidal gene cry1Ah was cloned from Bacillus thuringiensis isolate BT8 previously for plant genetic engineering improvement. Truncated active Cry1Ah toxin has a toxicity level similar to that of the full-length Cry1Ah toxin. In this study, plant expression vector pMhGM harboring truncated cry1Ah gene was transformed into maize (Zea mays L.) immature embryos by Agrobacterium tumefaciens-mediated transformation at which maize alcohol dehydrogenase matrix attachment regions (madMARs) were incorporated on both sides of the gene expression cassette to improve gene expression. A total of 23 PCR positive events were obtained with a transformation efficiency of 5% around. Bioassay results showed that events 1-4 and 1-5 exhibited enhanced resistance to the Asian corn borer (Ostrinia furnacalis). These two events were further confirmed by molecular analysis. Southern blot suggested that a single copy of the cry1Ah gene was successfully integrated into the maize genome. Western blot and ELISA showed that the foreign gene cry1Ah was expressed stably at high level in maize and could be inherited stably over generations. The results of a bioassay of T1-T4 transgenic maize plants indicated that the transgenic plants were highly toxic to the Asian corn borer and their resistance could be inherited stably from generation to generation. Thus, events 1-4 and 1-5 are good candidates for the breeding of insect-resistant maize.
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