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利用豆荚特异性和组成型启动子驱动的融合cry1Ab/Ac基因培育抗豆荚螟转基因鹰嘴豆。

Development of pod borer-resistant transgenic chickpea using a pod-specific and a constitutive promoter-driven fused cry1Ab/Ac gene.

作者信息

Ganguly Moumita, Molla Kutubuddin Ali, Karmakar Subhasis, Datta Karabi, Datta Swapan Kumar

机构信息

Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata, 700019, West Bengal, India.

出版信息

Theor Appl Genet. 2014 Dec;127(12):2555-65. doi: 10.1007/s00122-014-2397-5. Epub 2014 Sep 25.

Abstract

We studied pod-specific msg promoter from soybean and developed different transgenic lines of chickpea expressing fused cry1Ab/Ac constitutively and pod specifically for resistance against the destructive pest Helicoverpa armigera. Crystal (Cry) proteins derived from the soil bacterium Bacillus thuringiensis (Bt) play an important role in controlling infestation of Helicoverpa armigera, which has been considered a serious problem in chickpea productivity. This study was undertaken to overcome the problem by introducing fused cry1Ab/Ac insecticidal gene under the control of pod-specific soybean msg promoter as well as rice actin1 promoter into chickpea var. DCP 92-3 by Agrobacterium-mediated transformation. Transgenic chickpea lines were characterized by real-time PCR, ELISA and insect bioassay. Expression of fused cry gene under constitutive and pod-specific promoter results in increase of 77- and 110-fold, respectively, compared to non-transgenic control plants. Levels of Cry toxins produced under the control of actin1 and soybean msg promoter were also estimated by ELISA in the leaves and pods, respectively. The higher expression of fused cry gene caused a lethal effect in larvae. The results of insect bioassay study revealed significant reduction in the survival rate of H. armigera reared on transgenic chickpea twigs as well as on pods. Pod-specific promoter-driven fused cry gene provides better and significant management strategy of pest control of chickpea without phenotypic cost.

摘要

我们研究了大豆中豆荚特异性msg启动子,并培育了不同的鹰嘴豆转基因品系,这些品系组成型且豆荚特异性地表达融合的cry1Ab/Ac,以抵抗毁灭性害虫棉铃虫。来源于土壤细菌苏云金芽孢杆菌(Bt)的晶体(Cry)蛋白在控制棉铃虫侵害方面发挥着重要作用,棉铃虫一直被认为是鹰嘴豆产量的严重问题。本研究旨在通过农杆菌介导的转化,将融合的cry1Ab/Ac杀虫基因在豆荚特异性大豆msg启动子以及水稻肌动蛋白1启动子的控制下导入鹰嘴豆品种DCP 92 - 3,以克服这一问题。通过实时PCR、ELISA和昆虫生物测定对转基因鹰嘴豆品系进行了表征。与非转基因对照植株相比,组成型和豆荚特异性启动子控制下的融合cry基因表达分别增加了77倍和110倍。还分别通过ELISA在叶片和豆荚中估计了肌动蛋白1和大豆msg启动子控制下产生的Cry毒素水平。融合cry基因的高表达对幼虫产生了致死效应。昆虫生物测定研究结果显示,在转基因鹰嘴豆嫩枝和豆荚上饲养的棉铃虫存活率显著降低。豆荚特异性启动子驱动的融合cry基因提供了一种更好且显著的鹰嘴豆害虫防治管理策略,且无表型代价。

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