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FAR 基因敲除显著抑制二化螟的存活,且双链 FAR 在水稻中的表达具有很强的抗虫性。

FAR knockout significantly inhibits Chilo suppressalis survival and transgene expression of double-stranded FAR in rice exhibits strong pest resistance.

机构信息

Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, College of Plant Protection, Hunan Agricultural University, Changsha, China.

National Research Center of Engineering & Technology for Utilization of Botanical Functional Ingredients, Hunan Agricultural University, Changsha, China.

出版信息

Plant Biotechnol J. 2022 Dec;20(12):2272-2283. doi: 10.1111/pbi.13906. Epub 2022 Aug 26.

Abstract

Chilo suppressalis is one of the most prevalent and damaging rice pests, causing significant economic losses each year. Chemical control is currently the primary method of controlling C. suppressalis. However, the indiscriminate use of chemical insecticides increases pest resistance, pollutes the environment and poses a significant health threat to humans and livestock, highlighting the need to find safer, more pest-specific and more effective alternatives to pest control. Plant-mediated RNA interference (RNAi) is a promising agricultural pest control method that is highly pest-specific and has less of an impact on the environment. Using multi-sgRNAs/Cas9 technology to delete Fatty acyl-CoA reductase (FAR) of C. suppressalis in the G0 generation, we show that downregulating FAR transcription may significantly increase the mortality rate and darken the epidermis of C. suppressalis compared with the control. Subsequently, we developed dsFAR transgenic rice lines using Agrobacterium-mediated genetic transformation and then screened three strains expressing dsFAR at high levels using transcriptional level analysis. Using transgenic rice stems, a laboratory feeding bioassay indicated that at least one line (L#10) displayed a particularly high level of insect resistance, with an insect mortality rate of more than 80%. In the field trials, dsFAR transgenic rice displayed high levels of resistance to C. suppressalis damage. Collectively, these results suggest the potential of a new environment-friendly, species-specific strategy for rice pest management.

摘要

二化螟是一种常见且具有破坏性的水稻害虫,每年都会造成重大经济损失。目前,化学防治仍是防治二化螟的主要方法。然而,化学杀虫剂的滥用会导致害虫产生抗药性,污染环境,对人类和牲畜的健康构成严重威胁,因此需要寻找更安全、更具针对性、更有效的害虫防治替代方法。植物介导的 RNA 干扰 (RNAi) 是一种很有前途的农业害虫防治方法,它具有高度的针对性,对环境的影响较小。本研究利用多 sgRNAs/Cas9 技术在 G0 代敲除二化螟的 Fatty acyl-CoA reductase (FAR),结果表明,与对照相比,下调 FAR 转录可能会显著增加二化螟的死亡率并使表皮黑化。随后,我们利用农杆菌介导的遗传转化技术构建了 dsFAR 转基因水稻株系,并通过转录水平分析筛选出 3 株高表达 dsFAR 的株系。利用转基因水稻茎进行的室内饲养生物测定表明,至少有一个株系(L#10)表现出特别高的抗虫性,昆虫死亡率超过 80%。在田间试验中,dsFAR 转基因水稻对二化螟的危害表现出高抗性。综上所述,这些结果表明,针对水稻害虫管理的一种新的环保、物种特异性策略具有潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6e27/11383565/af8fb1e7ae96/PBI-20-2272-g005.jpg

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