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一个赋予棉花植株对草地贪夜蛾和棉铃象甲抗性的cry1Ia基因的稳定整合与表达。

Stable integration and expression of a cry1Ia gene conferring resistance to fall armyworm and boll weevil in cotton plants.

作者信息

Silva Carliane Rc, Monnerat Rose, Lima Liziane M, Martins Érica S, Melo Filho Péricles A, Pinheiro Morganna Pn, Santos Roseane C

机构信息

Federal Rural University of Pernambuco, Dois Irmãos, Recife, PE, Brazil.

Embrapa - Genetic Resources and Biotechnology (CENARGEN), SAIN, Brasília, DF, Brazil.

出版信息

Pest Manag Sci. 2016 Aug;72(8):1549-57. doi: 10.1002/ps.4184. Epub 2015 Dec 18.

Abstract

BACKGROUND

Boll weevil is a serious pest of cotton crop. Effective control involves applications of chemical insecticides, increasing the cost of production and environmental pollution. The current genetically modified Bt crops have allowed great benefits to farmers but show activity limited to lepidopteran pests. This work reports on procedures adopted for integration and expression of a cry transgene conferring resistance to boll weevil and fall armyworm by using molecular tools.

RESULTS

Four Brazilian cotton cultivars were microinjected with a minimal linear cassette generating 1248 putative lines. Complete gene integration was found in only one line (T0-34) containing one copy of cry1Ia detected by Southern blot. Protein was expressed in high concentration at 45 days after emergence (dae), decreasing by approximately 50% at 90 dae. Toxicity of the cry protein was demonstrated in feeding bioassays revealing 56.7% mortality to boll weevil fed buds and 88.1% mortality to fall armyworm fed leaves. A binding of cry1Ia antibody was found in the midgut of boll weevils fed on T0-34 buds in an immunodetection assay.

CONCLUSION

The gene introduced into plants confers resistance to boll weevil and fall armyworm. Transmission of the transgene occurred normally to T1 progeny. All plants showed phenotypically normal growth, with fertile flowers and abundant seeds. © 2015 Society of Chemical Industry.

摘要

背景

棉铃象鼻虫是棉花作物的一种严重害虫。有效的防治措施包括施用化学杀虫剂,这增加了生产成本并造成环境污染。当前的转基因Bt作物给农民带来了巨大益处,但显示出的活性仅限于鳞翅目害虫。这项工作报道了通过使用分子工具将赋予对棉铃象鼻虫和草地贪夜蛾抗性的cry转基因整合并表达所采用的程序。

结果

对四个巴西棉花品种进行了微量注射,使用一个最小线性盒构建了1248个推定株系。通过Southern印迹检测发现,仅在一个株系(T0 - 34)中发现了完整的基因整合,该株系含有一个cry1Ia拷贝。在出苗后45天(dae)蛋白质以高浓度表达,在90 dae时下降约50%。在喂食生物测定中证明了cry蛋白的毒性,结果显示以T0 - 34花蕾为食的棉铃象鼻虫死亡率为56.7%,以叶片为食的草地贪夜蛾死亡率为88.1%。在免疫检测试验中,在以T0 - 34花蕾为食的棉铃象鼻虫中肠中发现了cry1Ia抗体结合。

结论

导入植物中的基因赋予了对棉铃象鼻虫和草地贪夜蛾的抗性。转基因正常传递到T1代后代。所有植株在表型上生长正常,花朵可育且种子丰富。© 2015化学工业协会。

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