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在从芦笋中分离出的创伤诱导型启动子(AoPR1)控制下,Cry1Ac在转基因烟草植株中的表达,用于防治烟草天蛾和烟草夜蛾。

Expression of Cry1Ac in transgenic tobacco plants under the control of a wound-inducible promoter (AoPR1) isolated from Asparagus officinalis to control Heliothis virescens and Manduca sexta.

作者信息

Gulbitti-Onarici Selma, Zaidi Mohsin Abbas, Taga Ibrahim, Ozcan Sebahattin, Altosaar Illimar

机构信息

Department of Biochemistry Microbiology & Immunology, University of Ottawa, Ottawa, ON, K1H 8M5, Canada.

出版信息

Mol Biotechnol. 2009 Jul;42(3):341-9. doi: 10.1007/s12033-009-9168-6. Epub 2009 Apr 8.

Abstract

Expression of cry1Ac gene from Bacillus thuringiensis (Bt) was evaluated under the control of a wound-inducible AoPR1 promoter from Asparagus officinalis in transgenic tobacco plants. The leaves of transgenic plants were mechanically wounded to evaluate the activity of the AoPR1 promoter in driving the expression of Cry1Ac protein at the wound site. Our results indicate that mechanical wounding of transgenic plants was effective in inducing the expression of Cry1Ac protein. As a result of this induction, the accumulated levels of Cry1Ac protein increased during 6-72 h post-wounding period. The leaves of transgenic tobacco plants were evaluated for resistance against Heliothis virescens and Manduca sexta in insect bioassays in two different ways. The detached tobacco leaves were either fed directly to the insect larvae or they were first mechanically wounded followed by a 72 h post-wounding feeding period. Complete protection of mechanically wounded leaves of transgenic plants was observed within 24 h of the bioassay. The leaves of transgenic plants fed directly (without pre-wounding) to the larvae achieved the same level of protection between 24 and 72 h of the bioassay.

摘要

在来自芦笋的伤口诱导型AoPR1启动子的控制下,评估了苏云金芽孢杆菌(Bt)cry1Ac基因在转基因烟草植株中的表达。对转基因植株的叶片进行机械损伤,以评估AoPR1启动子驱动伤口部位Cry1Ac蛋白表达的活性。我们的结果表明,转基因植株的机械损伤有效地诱导了Cry1Ac蛋白的表达。由于这种诱导作用,在损伤后6至72小时内,Cry1Ac蛋白的积累水平有所增加。通过两种不同方式在昆虫生物测定中评估了转基因烟草植株叶片对棉铃虫和烟草天蛾的抗性。将分离的烟草叶片直接喂给昆虫幼虫,或者先对其进行机械损伤,然后在损伤后72小时的喂食期进行喂食。在生物测定的24小时内观察到转基因植株机械损伤叶片得到了完全保护。直接(无预先损伤)喂给幼虫的转基因植株叶片在生物测定的24至72小时内达到了相同的保护水平。

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