Full Text:   <1875>

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CLC number: Q182

On-line Access: 2015-10-03

Received: 2015-03-09

Revision Accepted: 2015-06-09

Crosschecked: 2015-09-13

Cited: 0

Clicked: 3193

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Qi-chao Zhao

http://orcid.org/0000-0001-5975-4489

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Journal of Zhejiang University SCIENCE B 2015 Vol.16 No.10 P.824-831

http://doi.org/10.1631/jzus.B1500056


Generation of insect-resistant and glyphosate-tolerant rice by introduction of a T-DNA containing two Bt insecticidal genes and an EPSPS gene


Author(s):  Qi-chao Zhao, Ming-hong Liu, Xian-wen Zhang, Chao-yang Lin, Qing Zhang, Zhi-cheng Shen

Affiliation(s):  State Key Laboratory of Rice Biology, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, China

Corresponding email(s):   zcshen@zju.edu.cn

Key Words:  Bt gene stacking, Bt resistance management, EPSPS, Transgenic rice


Qi-chao Zhao, Ming-hong Liu, Xian-wen Zhang, Chao-yang Lin, Qing Zhang, Zhi-cheng Shen. Generation of insect-resistant and glyphosate-tolerant rice by introduction of a T-DNA containing two Bt insecticidal genes and an EPSPS gene[J]. Journal of Zhejiang University Science B, 2015, 16(10): 824-831.

@article{title="Generation of insect-resistant and glyphosate-tolerant rice by introduction of a T-DNA containing two Bt insecticidal genes and an EPSPS gene",
author="Qi-chao Zhao, Ming-hong Liu, Xian-wen Zhang, Chao-yang Lin, Qing Zhang, Zhi-cheng Shen",
journal="Journal of Zhejiang University Science B",
volume="16",
number="10",
pages="824-831",
year="2015",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1500056"
}

%0 Journal Article
%T Generation of insect-resistant and glyphosate-tolerant rice by introduction of a T-DNA containing two Bt insecticidal genes and an EPSPS gene
%A Qi-chao Zhao
%A Ming-hong Liu
%A Xian-wen Zhang
%A Chao-yang Lin
%A Qing Zhang
%A Zhi-cheng Shen
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%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1500056

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T1 - Generation of insect-resistant and glyphosate-tolerant rice by introduction of a T-DNA containing two Bt insecticidal genes and an EPSPS gene
A1 - Qi-chao Zhao
A1 - Ming-hong Liu
A1 - Xian-wen Zhang
A1 - Chao-yang Lin
A1 - Qing Zhang
A1 - Zhi-cheng Shen
J0 - Journal of Zhejiang University Science B
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PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B1500056


Abstract: 
Insect resistance and glyphosate tolerance have been two of the most important traits in the genetic improvement of various crops. In this study, two Bacillus thuringiensis (Bt) insecticidal genes, Cry1Ac and Cry1Ig, and a modified glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene (G10) were combined into a single transferred DNA (T-DNA) fragment and introduced into rice by Agrobacterium-mediated transformation. A transgenic line with single-copy T-DNA insertion named GAI-14 was found to be highly resistant to striped stem borer and rice leaf roller, and tolerant to glyphosate. Analysis of T-DNA border sequence suggested that the transgenes were inserted at the chromosome 3 and appeared to have not interrupted any known or putative genes. A field trial observed no significant difference in the basic agronomic traits between GAI-14 and the recipient rice.

转Cry1Ac、Cry1Ig和EPSPS基因水稻及其抗虫、抗草甘膦活性鉴定

目的:获得具有高水平表达Cry1AcCry1IgEPSPS基因并且具有抗虫、抗草甘膦活性的转基因水稻株。
创新点:Cry1Ig为新发现的苏云金杆菌(Bt)杀虫基因;Cry1IgCry1Ac共同使用有利于延缓害虫产生对Bt杀虫蛋白抗性。
方法:Cry1AcCry1IgEPSPSG10)基因的表达框依次插入到同一转移DNA(T-DNA)中,再将此T-DNA利用农杆菌介导法转化到水稻中。从转基因后代中,利用蛋白质印迹法(Western blot)、DNA印迹法(Southern blot)和热不对称交错聚合酶链式反应(TAIL-PCR)技术,筛选出能够高水平表达上述三个基因以及T-DNA单拷贝插入并且未发生明显插入突变的转化株。对于筛选到的转化株,利用Western blot进一步分析Cry1AcCry1IgG10基因的表达水平;使用酶联免疫吸附测定(ELISA)检测Cry1Ac的表达量;使用棉铃虫、二化螟和稻纵卷叶螟为对象,测定转基因水稻的抗虫活性;使用草甘膦喷施法,测定转基因水稻的抗草甘膦活性。最后,在大田试验中,考察转基因水稻与非转基因水稻的基本农艺性状,观察T-DNA的插入是否对转基因水稻的生长产生明显的影响。
结论:本实验最终获得具有高水平表达Cry1AcCry1IgG10基因以及T-DNA单拷贝插入并且未引起明显变化的转化株GAI-14。GAI-14对棉铃虫、二化螟、稻纵卷叶螟以及草甘膦均具有明显的抗性。田间试验表明GAI-14在基本农艺性状上与非转基因水稻无明显差异。

关键词:Bt基因堆叠;转基因水稻;EPSPS抗草甘膦基因;害虫抗性治理

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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