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利用转基因技术对功能验证秋黏虫 ABCC2 突变体对 Bt 产生抗性的影响。

Transgenic to Functionally Validate Fall Armyworm ABCC2 Mutations Conferring Bt Resistance.

机构信息

Department of Biology, University of Crete, Vassilika Vouton, 71409 Heraklion, Greece.

Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, 71409 Heraklion, Greece.

出版信息

Toxins (Basel). 2023 Jun 7;15(6):386. doi: 10.3390/toxins15060386.

Abstract

The fall armyworm (FAW), (J.E. Smith; Lepidoptera: Noctuidae) is an invasive agricultural pest with a global distribution, causing major crop losses annually. Its control strategies largely rely on chemical insecticides and transgenic crops expressing insecticidal proteins (Cry and Vip toxins); however, the development of high resistance poses a significant issue. The ATP-binding cassette transporter C2 (ABCC2) has been linked to Cry toxin pore formation, acting as a receptor of some Cry toxins. Recently detected mutations in the SfABCC2 gene in extracellular loop 4 (ECL4) have been associated with Bt toxin resistance in FAW. In the present study, we expressed the SfABCC2 gene in , a species normally unaffected by the Bt toxins. We demonstrate that susceptibility can be introduced by the ectopic and tissue-specific expression of wildtype SfABCC2. Next, we introduced mutations into ECL4-both individually and in combination-that have been recently described in Brazilian FAW and functionally validated by toxicity bioassays against the foliar Bt product Xentari. Our results provide an efficient demonstration of the suitability of transgenic for validating FAW ABCC2 resistance mutations in ECL4 against Bt toxins, and potential cross-resistance issues between closely related proteins that use ABCC2.

摘要

秋粘虫(FAW),(J.E. Smith;鳞翅目:夜蛾科)是一种具有全球分布的入侵性农业害虫,每年造成重大作物损失。其控制策略主要依赖于化学杀虫剂和表达杀虫蛋白(Cry 和 Vip 毒素)的转基因作物;然而,高抗性的发展是一个重大问题。ATP 结合盒转运蛋白 C2(ABCC2)与 Cry 毒素孔形成有关,作为一些 Cry 毒素的受体。最近在 SfABCC2 基因的细胞外环 4(ECL4)中检测到的突变与 FAW 中的 Bt 毒素抗性有关。在本研究中,我们在 中表达了 SfABCC2 基因,该物种通常不受 Bt 毒素的影响。我们证明,通过野生型 SfABCC2 的异位和组织特异性表达,可以引入敏感性。接下来,我们引入了 ECL4 中的突变-单独引入和组合引入-这些突变最近在巴西 FAW 中被描述,并通过针对叶面 Bt 产品 Xentari 的毒性生物测定进行了功能验证。我们的结果提供了一个有效的证明,即转基因 适合验证 ECL4 中针对 Bt 毒素的 FAW ABCC2 抗性突变,以及使用 ABCC2 的密切相关蛋白之间的潜在交叉抗性问题。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e280/10301211/cacf1933b41a/toxins-15-00386-g001.jpg

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