Non-transgenic, PAMAM co-delivery DNA of interactive proteins NbCRVP and NbCalB endows Nicotiana benthamiana with a stronger antiviral effect to RNA viruses

文献类型： 外文期刊

第一作者： Song, Liyun

作者： Song, Liyun;Shen, Lili;Li, Ying;Wang, Fenglong;Jiao, Yubing;Yang, Jinguang;Song, Liyun;Zhang, Daoshun;Liu, Tianbo;Jiang, Changqing;Li, Bin;Li, Changquan

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关键词： Cysteine-rich venom protein; Calcium-dependent lipid-binding (CaLB domain) family protein; Poly(amidoamine) (PAMAM); PAMAM@NbCRVP:NbCalB; Broad-spectrum antiviral effect

期刊名称：JOURNAL OF NANOBIOTECHNOLOGY （ 影响因子：10.2； 五年影响因子：11.5 ）

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年卷期： 2024 年 22 卷 1 期

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收录情况： SCI

摘要： Background Viral diseases continue to pose a major threat to the world's commercial crops. The in-depth exploration and efficient utilization of resistance proteins have become crucial strategies for their control. However, current delivery methods for introducing foreign DNA suffer from host range limitations, low transformation efficiencies, tissue damage, or unavoidable DNA integration into the host genome. The nanocarriers provides a convenient channel for the DNA delivery and functional utilization of disease-resistant proteins.Results In this research, we identified a cysteine-rich venom protein (NbCRVP) in Nicotiana benthamiana for the first time. Virus-induced gene silencing and transient overexpression clarified that NbCRVP could inhibit the infection of tobacco mosaic virus, potato virus Y, and cucumber mosaic virus, making it a broad-spectrum antiviral protein. Yeast two-hybrid assay, co-immunoprecipitation, and bimolecular fluorescence complementation revealed that calcium-dependent lipid-binding (CaLB domain) family protein (NbCalB) interacted with NbCRVP to assist NbCRVP playing a stronger antiviral effect. Here, we demonstrated for the first time the efficient co-delivery of DNA expressing NbCRVP and NbCalB into plants using poly(amidoamine) (PAMAM) nanocarriers, achieving stronger broad-spectrum antiviral effects.Conclusions Our work presents a tool for species-independent transfer of two interacting protein DNA into plant cells in a specific ratio for enhanced antiviral effect without transgenic integration, which further demonstrated new strategies for nanocarrier-mediated DNA delivery of disease-resistant proteins.

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