Silencing Arbuscular Mycorrhizal Fungal Gene Using Chitosan Nanoparticle-Mediated dsRNA Delivery System.

IF 1 Q3 BIOLOGY
Chumei Yan, Yuemin Wang, Qianhuang Guo, Hongjuan Huan, Siwei Wang, Xiaoning Fan, Xianan Xie
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Abstract

It has been discovered that many phytopathogenic fungi can absorb exogenous double-stranded RNAs (dsRNAs) to silence target genes, inhibiting fungal growth and pathogenicity for plant protection. In our recent report, the beneficial arbuscular mycorrhizal (AM) fungi are capable of acquiring external naked dsRNAs; however, whether the dsRNAs can be delivered into AM fungi through nanocarriers remains to be investigated. Here, we introduce a simple and advanced method for in vitro synthesizing chitosan (CS)/dsRNA polyplex nanoparticles (PNs) to silence the target gene in the AM fungus Rhizophagus irregularis. This method is straightforward, requiring minimal modifications, and is both efficient and eco-friendly, offering potential for rapid application in elucidating gene functions in AM fungi. Key features • The chitosan can carry the dsRNA derived from the AM fungus Rhizophagus irregularis. • CS/dsRNA polyplex nanoparticles (PNs) can successfully silence the target gene in the AM fungus R. irregularis. • CS/dsRNA PNs can be applied to the characterization of AM fungal genes via the spray-induced gene silencing (SIGS) approach. • This protocol can be applied in asymbiotic and symbiotic cultures of AM fungi. Graphical overview Overview of the chitosan/dsRNA gene silencing procedures.

壳聚糖纳米颗粒介导的dsRNA传递系统沉默丛枝菌根真菌基因。
研究发现,许多植物病原真菌可吸收外源双链rna (dsRNAs)沉默靶基因,抑制真菌生长和致病性,达到保护植物的目的。在我们最近的报告中,有益的丛枝菌根(AM)真菌能够获得外部裸dsRNAs;然而,dsrna是否可以通过纳米载体传递到AM真菌中仍有待研究。本文介绍了一种简单、先进的体外合成壳聚糖(CS)/dsRNA复合纳米颗粒(PNs)的方法,以沉默AM真菌不规则根噬菌(Rhizophagus irregularis)的靶基因。该方法简单、修改少、高效、环保,具有快速应用于AM真菌基因功能研究的潜力。•壳聚糖可携带AM真菌Rhizophagus irregularis的dsRNA。CS/dsRNA复合纳米颗粒(PNs)可以成功地沉默AM真菌中的靶基因。CS/dsRNA PNs可以通过喷雾诱导基因沉默(SIGS)方法应用于AM真菌基因的表征。•该方案可应用于AM真菌的非共生和共生培养。图解概述壳聚糖/dsRNA基因沉默程序概述。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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CiteScore
1.50
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0.00%
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