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MiDaf16 样和 MiSkn1 样基因家族是开发用于控制和管理南方根结线虫的生物技术工具的可靠靶标。

MiDaf16-like and MiSkn1-like gene families are reliable targets to develop biotechnological tools for the control and management of Meloidogyne incognita.

机构信息

Embrapa Genetic Resources and Biotechnology, Brasília-DF, 70297-400, Brazil.

Federal University of Brasília, Brasília-DF, 70910-900, Brazil.

出版信息

Sci Rep. 2020 Apr 24;10(1):6991. doi: 10.1038/s41598-020-63968-8.

Abstract

Meloidogyne incognita is a plant-parasitic root-knot nematode (RKN, PPN) responsible for causing damage to several crops worldwide. In Caenorhabditis elegans, the DAF-16 and SKN-1 transcription factors (TFs) orchestrate aging, longevity, and defense responses to several stresses. Here, we report that MiDaf16-like1 and MiSkn1-like1, which are orthologous to DAF-16 and SKN-1 in C. elegans, and some of their targets, are modulated in M. incognita J2 during oxidative stress or plant parasitism. We used RNAi technology for the stable production of siRNAs in planta to downregulate the MiDaf16-like1 and MiSkn1-like1 genes of M. incognita during host plant parasitism. Arabidopsis thaliana and Nicotiana tabacum overexpressing a hairpin-derived dsRNA targeting these genes individually (single-gene silencing) or simultaneously (double-gene silencing) were generated. T plants were challenged with M. incognita and the number of eggs, galls, and J2, and the nematode reproduction factor (NRF) were evaluated. Our data indicate that MiDaf16-like1, MiSkn1-like1 and some genes from their networks are modulated in M. incognita J2 during oxidative stress or plant parasitism. Transgenic A. thaliana and N. tabacum plants with single- or double-gene silencing showed significant reductions in the numbers of eggs, J2, and galls, and in NRF. Additionally, the double-gene silencing plants had the highest resistance level. Gene expression assays confirmed the downregulation of the MiDaf16-like1 and MiSkn1-like1 TFs and defense genes in their networks during nematode parasitism in the transgenic plants. All these findings demonstrate that these two TFs are potential targets for the development of biotechnological tools for nematode control and management in economically important crops.

摘要

南方根结线虫是一种植物寄生性根结线虫(RKN,PPN),可导致全球多种作物受损。在秀丽隐杆线虫中,DAF-16 和 SKN-1 转录因子(TFs)协调衰老、长寿和对多种应激的防御反应。在这里,我们报告说,MiDaf16-like1 和 MiSkn1-like1,这是秀丽隐杆线虫中 DAF-16 和 SKN-1 的同源物,以及它们的一些靶标,在氧化应激或植物寄生时在南方根结线虫 J2 中被调节。我们使用 RNAi 技术在植物体内稳定产生 siRNA,以在宿主植物寄生时下调南方根结线虫的 MiDaf16-like1 和 MiSkn1-like1 基因。生成了拟南芥和烟草过表达针对这些基因的发夹衍生 dsRNA 的转基因植物(单个基因沉默)或同时(双基因沉默)。用 M. incognita 挑战 T 植物,评估卵、虫瘿和 J2 的数量以及线虫繁殖因子(NRF)。我们的数据表明,MiDaf16-like1、MiSkn1-like1 和它们网络中的一些基因在 M. incognita J2 中在氧化应激或植物寄生时被调节。具有单基因或双基因沉默的转基因拟南芥和烟草植物在卵、J2 和虫瘿的数量以及 NRF 方面显示出显著减少。此外,双基因沉默植物具有最高的抗性水平。基因表达分析证实了在转基因植物中,线虫寄生时 MiDaf16-like1 和 MiSkn1-like1 TFs 和防御基因在它们的网络中的下调。所有这些发现表明,这两个 TF 是开发生物技术工具以控制和管理经济上重要作物中的线虫的潜在靶标。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1f99/7181638/a7d8ad4ae8de/41598_2020_63968_Fig1_HTML.jpg

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