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在植物体内表达双链 RNA 实现真菌细胞中转基因的体内特异性基因沉默

In vivo trans-specific gene silencing in fungal cells by in planta expression of a double-stranded RNA.

机构信息

Embrapa Recursos Genéticos e Biotecnologia, PqEB W5 Norte, 70770-900, Brasília, DF, Brazil.

出版信息

BMC Biol. 2010 Mar 31;8:27. doi: 10.1186/1741-7007-8-27.

Abstract

BACKGROUND

Self-complementary RNA transcripts form a double-stranded RNA (dsRNA) that triggers a sequence-specific mRNA degradation, in a process known as RNA interference (RNAi), leading to gene silencing. In vascular plants, RNAi molecules trafficking occur between cells and systemically throughout the plant. RNAi signals can spread systemically throughout a plant, even across graft junctions from transgenic to non-transgenic stocks. There is also a great interest in applying RNAi to pathogenic fungi. Specific inhibition of gene expression by RNAi has been shown to be suitable for a multitude of phytopathogenic filamentous fungi. However, double-stranded (ds)RNA/small interfering (si)RNA silencing effect has not been observed in vivo.

RESULTS

This study demonstrates for the first time the in vivo interference phenomenon in the pathogenic fungus Fusarium verticillioides, in which expression of an individual fungal transgene was specifically abolished by inoculating mycelial cells in transgenic tobacco plants engineered to express siRNAs from a dsRNA corresponding to the particular transgene.

CONCLUSION

The results provide a powerful tool for further studies on molecular plant-microbe and symbiotic interactions. From a biotechnological perspective, silencing of fungal genes by generating siRNAs in the host provides a novel strategy for the development of broad fungi-resistance strategies in plants and other organisms.

摘要

背景

自我互补的 RNA 转录本形成双链 RNA(dsRNA),在 RNA 干扰(RNAi)过程中触发特定序列的 mRNA 降解,导致基因沉默。在维管束植物中,RNAi 分子在细胞之间以及整个植物中进行细胞间运输。RNAi 信号可以在植物体内系统传播,甚至可以跨越转基因和非转基因品系的嫁接接头。将 RNAi 应用于致病真菌也引起了人们的极大兴趣。通过 RNAi 特异性抑制基因表达已被证明适用于多种植物病原丝状真菌。然而,双链 (ds)RNA/小干扰 (si)RNA 沉默效应尚未在体内观察到。

结果

本研究首次证明了在致病真菌轮枝镰孢中存在体内干扰现象,通过将表达 dsRNA 的菌丝细胞接种到转基因烟草植物中,可以特异性地消除个别真菌转基因的表达,dsRNA 对应于特定的转基因。

结论

这些结果为进一步研究分子植物-微生物和共生相互作用提供了有力的工具。从生物技术的角度来看,通过在宿主中产生 siRNA 来沉默真菌基因为在植物和其他生物中开发广谱真菌抗性策略提供了一种新策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a54e/2907587/00f279120f27/1741-7007-8-27-1.jpg

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