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Targeting chitinase gene of Helicoverpa armigera by host-induced RNA interference confers insect resistance in tobacco and tomato

机译:宿主诱导的RNA干扰靶向棉铃虫的几丁质酶基因赋予烟草和番茄抗虫性

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摘要

Helicoverpa armigera Hubner (Lepidoptera: Noctuidae) is a devastating agricultural insect pest with broad spectrum of host range, causing million dollars crop loss annually. Limitations in the present conventional and transgenic approaches have made it crucial to develop sustainable and environmental friendly methods for crop improvement. In the present study, host-induced RNA interference (HI-RNAi) approach was used to develop H. armigera resistant tobacco and tomato plants. Chitinase (HaCHI) gene, critically required for insect molting and metamorphosis was selected as a potential target. Hair-pin RNAi construct was prepared from the conserved off-target free partial HaCHI gene sequence and was used to generate several HaCHI-RNAi tobacco and tomato plants. Northern hybridization confirmed the production of HaCHI gene-specific siRNAs in HaCHI-RNAi tobacco and tomato lines. Continuous feeding on leaves of RNAi lines drastically reduced the target gene transcripts and consequently, affected the overall growth and survival of H. armigera. Various developmental deformities were also manifested in H. armigera larvae after feeding on the leaves of RNAi lines. These results demonstrated the role of chitinase in insect development and potential of HI-RNAi for effective management of H. armigera.
机译:棉铃虫Hubner(鳞翅目:夜蛾科)是具有毁灭性的农业害虫,具有广泛的寄主范围,每年造成数百万美元的作物损失。当前常规和转基因方法的局限性使得开发可持续和环境友好的作物改良方法变得至关重要。在本研究中,使用宿主诱导的RNA干扰(HI-RNAi)方法开发抗棉铃虫的烟草和番茄植物。昆虫蜕皮和变态反应至关重要的几丁质酶(HaCHI)基因被选为潜在目标。从保守的脱靶游离HaCHI部分基因序列制备发夹RNAi构建体,并将其用于生成几种HaCHI-RNAi烟草和番茄植物。北部杂交证实了在HaCHI-RNAi烟草和番茄品系中生产了HaCHI基因特异性siRNA。连续取食RNAi系的叶子会极大地减少目标基因的转录本,因此影响棉铃虫的总体生长和存活。取食RNAi株系的叶片后,在棉铃虫幼虫中也表现出各种发育畸形。这些结果证明了几丁质酶在昆虫发育中的作用以及HI-RNAi在有效处理棉铃虫中的潜力。

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