Dataset Information

Construction of gold-siRNA _NPR1 nanoparticles for effective and quick silencing of NPR1 in Arabidopsis thaliana.

ABSTRACT: In recent years, gold nanoparticles (AuNPs) have been widely used as gene silencing agents and therapeutics for treatment of cancers due to their high transfection efficiency and lack of cytotoxicity, but their roles in gene silencing in plants have not yet been reported. Here, we report synthesis of AuNPs-branched polyethylenimine and its integration with the small interfering RNAs (siRNA) of NPR1 to form a AuNPs-siRNA _NPR1 compound. Our results showed that AuNPs-siRNA _NPR1 was capable of infiltrating into Arabidopsis cells. AuNPs-siRNA _NPR1 silenced 80% of the NPR1 gene in Arabidopsis. Bacteriostatic and ion leakage experiments suggest that the NPR1 gene in Arabidopsis leaves was silenced by AuNPs-siRNA _NPR1 . In Columbia-0 plants, compared with the control group treated with buffer solution, the AuNPs-siRNA _NPR1 treatment significantly increased the number of colonies and cell death, and the leaves turned yellow, similar to the phenotype of the npr1 leaves. These results indicated this AuNPs-siRNA _NPR1 silencing the NPR1 gene method is simple, effective and quick (3 days), and a powerful tool to study gene functions in plants.

SUBMITTER: Lei WX

PROVIDER: S-EPMC9054099 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Construction of gold-siRNA NPR1 nanoparticles for effective and quick silencing of NPR1 in Arabidopsis thaliana.

Lei Wen-Xue WX An Zi-Shuai ZS Zhang Bai-Hong BH Wu Qian Q Gong Wen-Jun WJ Li Jin-Ming JM Chen Wen-Li WL

RSC advances 20200520 33

In recent years, gold nanoparticles (AuNPs) have been widely used as gene silencing agents and therapeutics for treatment of cancers due to their high transfection efficiency and lack of cytotoxicity, but their roles in gene silencing in plants have not yet been reported. Here, we report synthesis of AuNPs-branched polyethylenimine and its integration with the small interfering RNAs (siRNA) of NPR1 to form a AuNPs-siRNA NPR1 compound. Our results showed that AuNPs-siRNA ...[more]

PMID: 35515443

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Project description:BackgroundAdvances in forward and reverse genetic techniques have enabled the discovery and identification of several plant defence genes based on quantifiable disease phenotypes in mutant populations. Existing models for testing the effect of gene inactivation or genes causing these phenotypes do not take into account eventual uncertainty of these datasets and potential noise inherent in the biological experiment used, which may mask downstream analysis and limit the use of these datasets. Moreover, elucidating biological mechanisms driving the induced disease resistance and influencing these observable disease phenotypes has never been systematically tackled, eliciting the need for an efficient model to characterize completely the gene target under consideration.ResultsWe developed a post-gene silencing bioinformatics (post-GSB) protocol which accounts for potential biases related to the disease phenotype datasets in assessing the contribution of the gene target to the plant defence response. The post-GSB protocol uses Gene Ontology semantic similarity and pathway dataset to generate enriched process regulatory network based on the functional degeneracy of the plant proteome to help understand the induced plant defence response. We applied this protocol to investigate the effect of the NPR1 gene silencing to changes in Arabidopsis thaliana plants following Pseudomonas syringae pathovar tomato strain DC3000 infection. Results indicated that the presence of a functionally active NPR1 reduced the plant's susceptibility to the infection, with about 99% of variability in Pseudomonas spore growth between npr1 mutant and wild-type samples. Moreover, the post-GSB protocol has revealed the coordinate action of target-associated genes and pathways through an enriched process regulatory network, summarizing the potential target-based induced disease resistance mechanism.ConclusionsThis protocol can improve the characterization of the gene target and, potentially, elucidate induced defence response by more effectively utilizing available phenotype information and plant proteome functional knowledge.

Dataset Information

Construction of gold-siRNA _NPR1 nanoparticles for effective and quick silencing of NPR1 in Arabidopsis thaliana.

Publications

Construction of gold-siRNA <sub><i>NPR1</i></sub> nanoparticles for effective and quick silencing of <i>NPR1</i> in <i>Arabidopsis thaliana</i>.

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